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Cristina MAGNONI

Professore Associato
Dipartimento Chirurgico, Medico, Odontoiatrico e di Scienze Morfologiche con interesse Trapiantologico, Oncologico e di Medicina Rigenerativa

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Pubblicazioni

- Nuovo sostituto dermico e sua applicazione terapeutica [Brevetto]
Giannetti, Alberto; Magnoni, Cristina; Benassi, Luisa; Pellegrini, Massimo
abstract

Il prodotto oggetto del brevetto è costituito da:a)cellule staminali/stromali multipotenti autologhe ottenute da microaspirazione di tessuto adiposo sottocutaneo e in grado di differenziare nelle cellule che costituiscono il derma umano e di produrre matrice connettivale; b) da una matrice biocompatibile tridimensionale.Le cellule stromali autologhe multipotenti sono seminate nello scaffold dopo un periodo di selezione-coltura in vitro. Questo prodotto è destinato alle strutture sanitarie specialistiche con le seguenti indicazioni terapeutiche: lesioni con perdita di tessuti molli di varia eziologia, sia di natura acuta (traumatiche, ustioni, esiti post-chirurgici) sia di natura cronica (ferite da pressione, ulcere vascolari, ulcere del piede diabetico).

2024 - Delay in cutaneous squamous cell carcinoma diagnosis due to interrupted services is associated with worse prognoses and modified surgical approaches [Articolo su rivista]
Taccioli, Filippo; Blessent, Claudio Gio Francesco; Paganelli, Alessia; Fagioli, Francesca; Chester, JOHANNA MARY; Kaleci, Shaniko; Costantini, Matteo; Barbara, Ferrari; Fiorentini, Chiara; DE SANTIS, Giorgio; Magnoni, Cristina
abstract

2023 - Allele specific CRISPR/Cas9 editing of dominant Epidermolysis Bullosa Simplex in human epidermal stem cells [Articolo su rivista]
Cattaneo, C; Enzo, E; De Rosa, L; Sercia, L; Consiglio, F; Forcato, M; Bicciato, S; Paiardini, A; Basso, G; Tagliafico, E; Paganelli, A; Fiorentini, C; Magnoni, C; Latella, M C; De Luca, M
abstract

: Epidermolysis Bullosa Simplex (EBS) is a rare skin disease inherited mostly in an autosomal dominant manner. Patients display a skin fragility that leads to blisters and erosions caused by minor mechanical trauma. EBS phenotypic and genotypic variants are caused by genetic defects in intracellular proteins whose function is to provide the attachment of basal keratinocytes to the basement membrane zone and most of EBS cases display mutations in keratin 5 (KRT5) and keratin 14 (KRT14) genes. Besides palliative treatments, there is still no long-lasting effective cure to correct the mutant gene and abolish dominant negative effect of the pathogenic protein over its wild-type counterpart. Here, we propose a molecular strategy for EBS01 patient's keratinocytes carrying a monoallelic c.475/495del21 mutation in KRT14 exon1. Through the CRISPR/Cas9 system we performed a specific cleavage only on the mutant allele and restore a normal cellular phenotype and a correct intermediate filament network, without affecting the epidermal stem cell, referred to as holoclones, which play a crucial role in epidermal regeneration.

2023 - Comparative study of in-vitro autofluorescence of normal versus non-melanoma-skin-cancer cells at different excitation wavelengths [Articolo su rivista]
Garbarino, Federico; Scelfo, Daniel; Paulone, Gabriele; Paganelli, Alessia; Ulrici, Alessandro; Magnoni, Cristina; Pasquali, Luca
abstract

: In this experimental study the autofluorescence of squamous carcinoma cells, stimulated by 6 different excitation wavelengths in the range 280-533 nm, has been compared with the autofluorescence of normal control keratinocytes. Skin cells were cultivated in vitro, to isolate their characteristic autofluorescence form the more complex one that would be originated by the complete skin tissue. Autofluorescence spectra in the visible range were complemented by absorption measurements. It was observed that the control cells showed characteristic emission (and absorption) structures due to typical endogenous chromophores [FAD and NAD(P)H, lipo-pigments, porphyrins], that were severely dumped in pathological cells. The autofluorescence spectra were then elaborated by multivariate analysis: after a first exploratory data analysis by means of Principal Component Analysis, the whole dataset was used to develop classification models using partial least squares-discriminant analysis, to differentiate between normal and pathological cells. This permitted us to identify the most suitable fluorescence spectral interval, in the 550-670 nm range, to discriminate between normal and pathological behavior, independently on the excitation wavelength.

2023 - Metabolic Profile of Whole Unstimulated Saliva in Patients with Sjögren's Syndrome [Articolo su rivista]
Setti, Giacomo; Righi, Valeria; Mucci, Adele; Panari, Lucia; Bernardelli, Giuditta; Tarentini, Elisabetta; Gambini, Anna; Consolo, Ugo; Generali, Luigi; Magnoni, Cristina; Meleti, Marco; Sandri, Gilda; Bellini, Pierantonio
abstract

: Primary Sjögren's Syndrome (pSS) is a multi-system autoimmune disease that involves the exocrine glands. Lymphocytes infiltrate the gland tissue, leading to anatomical modification and hypofunction. Even if the prognosis of pSS is favorable, quality of life is typically reduced due to the diverse manifestations of the disease. The aim of this study is to compare the salivary metabolomes of pSS with healthy controls (HCs). Seven cases were selected from a cohort of pSS patients, and six age- and sex-matched HCs were recruited from a cohort of volunteers. Whole unstimulated saliva was collected for NMR analysis. Our metabolomic analysis focused on 360 ms total echo 1D 1H NMR CPMG spectra. Metabolites detected with CPMG NMR spectra were assigned through 2D NMR spectra (COSY, TOCSY, and HSQC). About 50 metabolites were detected and assigned. Unsupervised exploratory PCA returned partial clustering, and PLS-DA improved the separation between pSS and HCs, highlighting a pool of metabolites distinctly describing each group. Despite the limited number of samples, the presented preliminary data are promising. PLS-DA indicated well-defined group separation, suggesting that the application of 1H-NMR metabolomics is suitable for the study of pSS.

2023 - NMR Metabolomics Based Study on Oral Malignant Disorders [Abstract in Atti di Convegno]
Gambini, A.; Setti, G.; Magnoni, C.; Meleti, M.; Mucci, A.; Righi, V.
abstract

2023 - Obesity and Wound Healing: Focus on Mesenchymal Stem Cells [Articolo su rivista]
Alma, Antonio; Marconi, Guya Diletta; Rossi, Elena; Magnoni, Cristina; Paganelli, Alessia
abstract

Chronic wounds represent nowadays a major challenge for both clinicians and researchers in the regenerative setting. Obesity represents one of the major comorbidities in patients affected by chronic ulcers and therefore diverse studies aimed at assessing possible links between these two morbid conditions are currently ongoing. In particular, adipose tissue has recently been described as having metabolic and endocrine functions rather than serving as a mere fat storage deposit. In this setting, adipose-derived stem cells, a peculiar subset of mesenchymal stromal/stem cells (MSCs) located in adipose tissue, have been demonstrated to possess regenerative and immunological functions with a key role in regulating both adipocyte function and skin regeneration. The aim of the present review is to give an overview of the most recent findings on wound healing, with a special focus on adipose tissue biology and obesity.

2023 - Researching the level of agreement among experts on terms used to describe wounds: An international study [Articolo su rivista]
Greco, Alessandro; Mastronicola, Diego; Pacini, Francesco; Giacomelli, LUCA MARIO; Papa, Simonetta; Fiorentini, Chiara; David, Valentina; WERTZBERGER ROWAN, Sara; Mennini, Natascia; Magnoni, Cristina
abstract

2023 - Wound Healing after Acellular Dermal Substitute Positioning in Dermato-Oncological Surgery: A Prospective Comparative Study [Articolo su rivista]
Paganelli, Alessia; Naselli, Andrea Giovanni; Bertoni, Laura; Rossi, Elena; Azzoni, Paola; Pisciotta, Alessandra; Cesinaro, Anna Maria; Benassi, Luisa; Kaleci, Shaniko; Garbarino, Federico; Ferrari, Barbara; Fiorentini, Chiara; Reggiani, Camilla; Magnoni, Cristina
abstract

Background: MatriDerm and Integra are both widely used collagenic acellular dermal matrices (ADMs) in the surgical setting, with similar characteristics in terms of healing time and clinical indication. The aim of the present study is to compare the two ADMs in terms of clinical and histological results in the setting of dermato-oncological surgery. Methods: Ten consecutive patients with medical indications to undergo surgical excision of skin cancers were treated with a 2-step procedure at our Dermatologic Surgery Unit. Immediately after tumor removal, both ADMs were positioned on the wound bed, one adjacent to the other. Closure through split-thickness skin grafting was performed after approximately 3 weeks. Conventional histology, immunostaining and ELISA assay were performed on cutaneous samples at different timepoints. Results: No significant differences were detected in terms of either final clinical outcomes or in extracellular matrix content of the neoformed dermis. However, Matriderm was observed to induce scar retraction more frequently. In contrast, Integra was shown to carry higher infectious risk and to be more slowly reabsorbed into the wound bed. Sometimes foreign body-like granulomatous reactions were also observed, especially in Integra samples. Conclusions: Even in the presence of subtle differences between the ADMs, comparable global outcomes were demonstrated after dermato-oncological surgery.

2022 - Atypical cellular neurothekeoma: report of a case [Articolo su rivista]
Pedroni, Gioia; Paganelli, Alessia; Bassoli, Sara; Kutzner, Heinz; Cesinaro, Anna Maria; Magnoni, Cristina
abstract

2022 - Current Knowledge in Skin Metabolomics: Updates from Literature Review [Articolo su rivista]
Paganelli, A.; Righi, V.; Tarentini, E.; Magnoni, C.
abstract

Metabolomic profiling is an emerging field consisting of the measurement of metabolites in a biological system. Since metabolites can vary in relation to different stimuli, specific metabolic patterns can be closely related to a pathological process. In the dermatological setting, skin metabolomics can provide useful biomarkers for the diagnosis, prognosis, and therapy of cutaneous disorders. The main goal of the present review is to present a comprehensive overview of the published studies in skin metabolomics. A search for journal articles focused on skin metabolomics was conducted on the MEDLINE, EMBASE, Cochrane, and Scopus electronic databases. Only research articles with electronically available English full text were taken into consideration. Studies specifically focused on cutaneous microbiomes were also excluded from the present search. A total of 97 papers matched all the research criteria and were therefore considered for the present work. Most of the publications were focused on inflammatory dermatoses and immune-mediated cutaneous disorders. Skin oncology also turned out to be a relevant field in metabolomic research. Only a few papers were focused on infectious diseases and rarer genetic disorders. All the major metabolomic alterations published so far in the dermatological setting are described extensively in this review.

2022 - In Vivo Melanoma Cell Morphology Reflects Molecular Signature and Tumor Aggressiveness [Articolo su rivista]
Marconi, Alessandra; Quadri, Marika; Farnetani, Francesca; Ciardo, Silvana; Palazzo, Elisabetta; Lotti, Roberta; Cesinaro, Anna Maria; Fabbiani, Luca; Vaschieri, Cristina; Puviani, Mario; Magnoni, Cristina; Kaleci, Shaniko; Pincelli, Carlo; Pellacani, Giovanni
abstract

Melanoma is the deadliest type of skin cancer, characterized by high cellular heterogeneity which contributes to therapy resistance and unpredictable disease outcome. Recently, by correlating Reflectance-Confocal-Microscopy (RCM) morphology with histopathological type, we identified four distinct melanoma-subtypes: dendritic-cell (DC), round-cell (RC), dermal-nest (DN), and combined-type (CT) melanomas. In the present study, each RCM-melanoma subtype expressed a specific biomolecular profile and biological behavior in vitro. Markers of tumor aggressiveness, including Ki67, MERTK, nestin and stemness markers, were highest in the most invasive CT and DN melanomas, as compared to DC and RC. This was also confirmed in multicellular tumor spheroids. Transcriptomic analysis showed a modulation of cancer progression-associated genes from DC to CT melanomas. The switch from E- to N-cadherin expression proved the epithelial-to-mesenchymal transition from DC to CT subtypes. The DN melanoma was predominantly located in the dermis, as also shown in skin reconstructs. It displayed a unique behavior and a molecular profile associated with a high degree of aggressiveness. Altogether, our results demonstrate that each RCM-melanoma subtype has a distinct biological and gene expression profile, related to tumor aggressiveness, confirming that RCM can be a dependable tool for in vivo detecting different types of melanoma and for early diagnostic screening.

2022 - Mesenchymal stromal cells promote the proliferation of basal stem cells and efficient epithelization in organotypic models of wound healing [Articolo su rivista]
Paganelli, A.; Benassi, L.; Rossi, E.; Tarentini, E.; Magnoni, C.
abstract

Adipose derived mesenchymal stromal cells (ADSCs) represent a fascinating tool in the scenario of wound healing and regenerative medicine. Recent data already demonstrated that ADSCs could exert a stimulatory action on epithelial cells through secretion of soluble factors. The aim of the present study was to assess how ADSCs guide wound re-epithelization in vitro in the presence of keratinocytes. We used an organotypic model of wound healing and we seeded keratinocytes on a ADSC-induced dermal matrix. Conventional hematoxylin–eosin stain and immunohistochemistry staining for Ki67, p63 and pan-keratins were performed at different timepoints. Histological sections of organotypic cultures showed complete coverage of the ADSC-induced matrix by keratinocytes. Proliferation of basal stem cells was found to be the main mechanism responsible for epithelization of the dermis. In conclusion, ADSC do not only stimulate dermal regeneration through collagen deposition but also promote epithelization.

2022 - Surgical management and oncological follow-up of cutaneous squamous cell carcinomas arising in epidermolysis bullosa patients [Articolo su rivista]
Paganelli, Alessia; Giordano, Erminia; Fiorentini, Chiara; Ferrari, Barbara; Reggiani, Camilla; Garbarino, Federico; Magnoni, Cristina
abstract

Background Hereditary epidermolysis bullosa (EB) is a rare genodermatosis characterized by skin fragility and blistering of the skin and mucous membranes in reaction to minimal traumas. The development of cutaneous squamous cell carcinomas (cSCCs) is one of the most common medical complications in junctional and dystrophic forms of the disease. Complete surgical excision of cutaneous tumors represents the gold standard of treatment. However, not only recognition of cSCCs can be challenging in the affected skin but also wound closure after surgical excision poses a great therapeutic challenge in EB patients. The aim of our study was to analyze the postoperative outcomes of such patients in order to have a better knowledge of the main critical issues in their surgical management and oncological follow-up. Methods We retrospectively identified a cohort of five EB patients treated at Modena University Hospital. Collected data included patient age and sex, date of cSCC diagnosis, relapses/recurrences, site of the neoplasm, number of surgical interventions, use of dermal substitutes, and postoperative infections. Results A total of 26 cSCCs were detected in our cohort. Forty-one surgical interventions were necessary to achieve excision of cSCCs with clear margins, varying from 1 to 4 surgical sessions per cSCC. Dermal substitutes were used in most cases but carried a higher infectious risk. Conclusions EB patients tend to develop numerous cSCCs that often relapse even after complete excision with clear margins. These results stress the importance of early cSCC diagnosis and strict postsurgical follow-up.

2022 - The Dark Side of Adipose-Derived Mesenchymal Stromal Cells in Cutaneous Oncology: Roles, Expectations, and Potential Pitfalls [Articolo su rivista]
Paganelli, Alessia; Rossi, Elena; Magnoni, Cristina
abstract

Adipose-derived stromal cells (ADSCs) have well-established regenerative and immunomodulatory properties. For such reasons, ADSCs are currently under investigation for their use in the setting of both regenerative medicine and autoimmune diseases. As per dermatological disorders, mesenchymal stromal cell (MSC)-based strategies represent potential therapeutic tools not only for chronic ulcers and wound healing, but also for immune-mediated dermatoses. However, a growing body of research has been focusing on the role of MSCs in human cancers, due to the potential oncological risk of using MSC-based strategies linked to their antiapoptotic, proangiogenic, and immunosuppressive properties. In the dermatological setting, ADSCs have shown not only to promote melanoma growth and invasiveness, but also to induce drug resistance. In contrast, genetically modified ADSCs have been demonstrated to efficiently target therapies at tumor sites, due to their migratory properties and their peculiar tropism for cancer microenvironment. The present review briefly summarizes the findings published so far on the use of ADSCs in the dermato-oncological setting, with the majority of data being available for melanoma.

2022 - Use of confocal microscopy imaging for in vitro assessment of adipose-derived mesenchymal stromal cells seeding on acellular dermal matrices: 3D reconstruction based on collagen autofluorescence [Articolo su rivista]
Paganelli, A.; Tarentini, E.; Benassi, L.; Scelfo, D.; Pisciotta, A.; Rossi, E.; Magnoni, C.
abstract

Background: Both mesenchymal stromal cells (MSCs) and acellular dermal matrices (ADMs) represent fascinating therapeutic tools in the wound healing scenario. Strategies aimed at combining these two treatment modalities are currently under investigation. Moreover, scarcity of quantitative, nondestructive techniques for quality assessment of engineered tissues poses great limitations in regenerative medicine and collagen autofluorescence-based imaging techniques are acquiring great importance in this setting. Objective: Our goals were to assess the in vitro interactions between ADSCs and ADMs and to analyze extracellular-matrix production. Methods: Adipose-derived MSCs (ADSC) were plated on 8-mm punch biopsies of a commercially available ADM (Integra®). Conventional histology with hematoxylin-eosin staining, environmental scanning electron microscopy, and confocal-laser scanning microscopy were used to obtain imaging of ADSC-seeded ADMs. Collagen production by ADSCs was quantified by mean fluorescence intensity (MFI), expressed in terms of positive pixels/field, obtained through ImageJ software processing of three-dimensional projections from confocal scanning images. Control conditions included: fibroblast-seeded ADM, ADSC- and fibroblast-induced scaffolds, and Integra® alone. Results: ADSCs were efficiently seeded on Integra® and were perfectly incorporated in the pores of the scaffold. Collagen production was revealed to be significantly higher when ADSCs were seeded on ADM rather than in all other control conditions. Collagen autofluorescence was efficiently used as a surrogate marker of ECM production. Conclusions: Combined therapies based on MSCs and collagenic ADMs are promising therapeutic options for chronic wounds. Not only ADSCs can be efficiently seeded on ADMs, but ADMs also seem to potentiate their regenerative properties, as highlightable from fluorescence confocal imaging.

2021 - Clinicopathological Features Associated with Fluorescence Alteration: Analysis of 108 Oral Malignant and Potentially Malignant Lesions [Articolo su rivista]
Giovannacci, I.; Magnoni, C.; Pedrazzi, G.; Vescovi, P.; Meleti, M.
abstract

Background: Several noninvasive diagnostic tools have been developed to aid the early detection of oral cancer and for evaluation before definitive biopsy. Among these, devices evaluating a tissues autofluorescence (AF) are emerging. In particular, the most well known of these is the VELscope® system (LED Medical Diagnostics, Inc., Barnaby, Canada), which emits a light of 400-460 nm. This study has been developed to describe the most relevant clinicopathological features associated with AF alterations in a set of patients with oral squamous cell carcinoma and potentially malignant disorders (PMDs). Materials and methods: Overall, 108 lesions from 60 patients with clinical diagnoses of potentially malignant oral disorders and carcinomas were included in the study. For each case, the following variables were recorded and compared with the AF pattern: (1) clinical appearance (white, red, and white/red); (2) histological diagnosis (no dysplasia, mild/moderate dysplasia, severe dysplasia/in situ carcinoma, invasive carcinoma, and verrucous carcinoma); and (3) clinicopathological diagnosis. Binomial logistic regression was performed to investigate whether clinical appearance and/or histological diagnosis were significant in determining the degree of AF. Results: Among the white lesions, 66% resulted in hyper-fluorescence, whereas the red lesions appeared hypo-fluorescent in 95.2% of cases. The AF was altered (both hypo-fluorescent and hyper-fluorescent) in 36% of lesions without dysplasia; in 75.9% of lesions with mild or moderate dysplasia and in the totality of the in situ, invasive, and verrucous carcinomas (p < 0.0001). With regard to the binomial logistic regression, variables were separately considered and both were extremely significant in determining the degree of AF. Conclusions: Promising evidence for the use of AF as an adjunctive tool to conventional oral examinations (COEs) has been demonstrated. However, although the sensitivity of AF examination associated with COE is very high, both the literature and this study agree to indicate a low specificity.

2021 - Correlation between autofluorescence intensity and histopathological features in non-melanoma skin cancer: An ex vivo study [Articolo su rivista]
Giovannacci, I.; Meleti, M.; Garbarino, F.; Cesinaro, A. M.; Mataca, E.; Pedrazzi, G.; Reggiani, C.; Paganelli, A.; Truzzi, A.; Elia, F.; Giacomelli, L.; Magnoni, C.
abstract

Non-melanoma skin cancer (NMSC) is the most common malignant tumor affecting fair-skinned people. Increasing incidence rates of NMSC have been reported worldwide, which is an important challenge in terms of public health management. Surgical excision with pre-operatively identified margins is one of the most common and effective treatment strategies. Incomplete tumor removal is associated with a very high risk of recurrence and re-excision. Biological tissues can absorb and re-emit specific light wave-lengths, detectable through spectrophotometric devices. Such a phenomenon is known as autofluorescence (AF). AF spectroscopy has been widely explored for non-invasive, early detection of NMSC as well as for evaluation of surgical margins before excision. Fluorescence-aided diagnosis is based on differences in spectral characteristics between healthy and neoplastic skin. Understanding the biological basis of such differences and correlating AF intensity to histological features could improve the diagnostic accuracy of skin fluorescence spectroscopy. The primary objective of the present pre-clinical ex vivo study is to investigate the correlation between the intensity of cutaneous AF and the histopathological features of NMSC. Ninety-eight lesions suggestive for NMSCs were radically excised from 75 patients (46 M; 29 F; mean age: 79 years). After removal, 115 specific reference points on lesions (“cases”; 59 on BBC, 53 on SCC and 3 on other lesions) and on peri-lesional healthy skin (controls; 115 healthy skin) were identified and marked through suture stitches. Such reference points were irradiated at 400–430 nm wavelength, and resulting emission AF spectra were acquired through spectrophotometry. For each case, AFIR (autofluorescence intensity ratio) was measured as the ratio between the number of photons emitted at a wavelength ranging between 450 and 700 nm (peak: 500 nm) in the healthy skin and that was captured in the pathological tissue. At the histological level, hyperkeratosis, neoangiogenesis, cellular atypia, epithelial thickening, fibrosis and elastosis were quantified by light microscopy and were assessed through a previously validated grading system. Statistical correlation between histologic variables and AFIR was calculated through linear regression. Spectrometric evaluation was performed on 230 (115 cases + 115 controls) reference points. The mean AFIR for BCC group was 4.5, while the mean AFIR for SCC group was 4.4 and the fluorescence peaks at 500 nm were approximately 4 times lower (hypo-fluorescent) in BCCs and in SCCs than in healthy skin. Histological variables significantly associated with alteration of AFIR were fibrosis and elastosis (p < 0.05), neoangiogenesis, hyperkeratosis and epithelial thickening. Cellular atypia was not significantly associated with alteration of AFIR. The intensity of fluorescence emission in neoplastic tissues was approximately 4 times lower than that in healthy tissues. Histopathological features such as hyperkeratosis, neoangiogenesis, fibrosis and elastosis are statistically associated with the decrease in AFIR. We hypothesize that such tissue alterations are among the possible biophysical and biochemical bases of difference in emission AF between neoplastic and healthy tissue. The results of the present evaluation highlighted the possible usefulness of autofluorescence as diagnostic, non-invasive and real-time tool for NMSCs.

2021 - Hologene 5: A Phase II/III Clinical Trial of Combined Cell and Gene Therapy of Junctional Epidermolysis Bullosa [Articolo su rivista]
De Rosa, L.; Enzo, E.; Zardi, G.; Bodemer, C.; Magnoni, C.; Schneider, H.; De Luca, M.
abstract

Epidermolysis bullosa (EB) is a group of devastating genetic diseases characterized by skin and mucosal fragility and formation of blisters, which develop either spontaneously or in response to minor mechanical trauma. There is no definitive therapy for any form of EB. Intermediate junctional EB (JEB) caused by mutations in the gene LAMB3 has been the first genetic skin disease successfully tackled by ex vivo gene therapy. Here, we present a multicenter, open-label, uncontrolled phase II/III study that aims at confirming the efficacy of Hologene 5, a graft consisting of cultured transgenic keratinocytes and epidermal stem cells and meant to combine cell and gene therapy for the treatment of LAMB3-related JEB. Autologous clonogenic keratinocytes will be isolated from patients’ skin biopsies, genetically corrected with a gamma-retroviral vector (γRV) carrying the full-length human LAMB3 cDNA and plated onto a fibrin support (144cm2). The transgenic epidermis will be transplanted onto surgically prepared selected skin areas of at least six JEB patients (four pediatric and two adults). Evaluation of clinical efficacy will include, as primary endpoint, a combination of clinical parameters, such as percentage of re-epithelialization, cellular, molecular, and functional parameters, mechanical stress tests, and patient-reported outcome (PRO), up to 12months after transplantation. Safety and further efficacy endpoints will also be assessed during the clinical trial and for additional 15years in an interventional non-pharmacological follow-up study. If successful, this clinical trial would provide a therapeutic option for skin lesions of JEB patients with LAMB3 mutations and pave the way to a combined cell and gene therapy platform tackling other forms of EB and different genodermatoses. Clinical Trial Registration: EudraCT Number: 2018-000261-36.

2021 - Integrated metabolomic analysis and cytokine profiling define clusters of immuno-metabolic correlation in new-onset psoriasis [Articolo su rivista]
Tarentini, Elisabetta; Odorici, Giulia; Righi, Valeria; Paganelli, Alessia; Giacomelli, Luca; Mirisola, Valentina; Mucci, Adele; Benassi, Luisa; D’Aversa, Elisabetta; Lasagni, Claudia; Kaleci, Shaniko; Reali, Eva; Magnoni, Cristina
abstract

The association between the metabolic profile and inflammatory cytokines in psoriasis is poorly understood. We analyzed the metabolic and cytokine/chemokine profiles in serum and skin from patients with new-onset psoriasis and healthy subjects (n = 7/group) by HR-MAS NMR and Bio-Plex immunoassay. Immuno-metabolic correlation matrix was analyzed in skin and serum to identify a potential immune-metabolic signature. Metabolomics analysis showed a significant increase in ascorbate and a decrease in scyllo-inositol, and a trend towards an increase in eight other metabolites in psoriatic skin. In serum, there was a significant increase of dimethylglycine and isoleucine. In parallel, psoriatic skin exhibited an increase of early inflammatory cytokines (IL-6, IL-8, TNF-α, IL-1β) and correlation analysis highlighted some major clusters of immune-metabolic correlations. A cluster comprising scyllo-inositol and lysine showed correlations with T-cell cytokines; a cluster comprising serine and taurine showed a negative correlation with early inflammatory cytokines (IL-6, G-CSF, CCL3). A strong positive correlation was enlightened between glutathione and inflammatory cytokines/angiogenesis promoters of psoriasis. The integration of metabolic and immune data indicated a molecular signature constituted by IL-6, IL1-ra, DMG, CCL4, Ile, Gly and IL-8, which could discriminate patients and healthy subjects and could represent a candidate tool in the diagnosis of new-onset psoriasis.

2021 - Metabolomic analysis of actinic keratosis and scc suggests a grade-independent model of squamous cancerization [Articolo su rivista]
Righi, V.; Reggiani, C.; Tarentini, E.; Mucci, A.; Paganelli, A.; Cesinaro, A. M.; Mataca, E.; Kaleci, S.; Ferrari, B.; Meleti, M.; Magnoni, C.
abstract

Background—Actinic keratoses (AKs) are the most common sun-induced precancerous lesions that can progress to squamocellular carcinoma (SCC). Recently, the grade-independent association between AKs and SCC has been suggested; however, the molecular bases of this potential association have not been investigated. This study has assessed the metabolomic fingerprint of AK I, AK II, AK III and SCC using high resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy in order to evaluate the hypothesis of grade-independent association between AK and SCC. Association between AKs and SCCs has also been evaluated by histopathology. Methods—Metabolomic data were obtained through HR-MAS NMR spectroscopy. The whole spectral profiles were analyzed through multivariate statistical analysis using MetaboAnalyst 5.0. Histologic examination was performed on sections stained with hematoxylin and eosin; statistical analysis was performed using STATA software version 14. Results—A group of 35 patients affected by AKs and/or SCCs and 10 healthy controls were enrolled for metabolomics analysis. Histopathological analysis was conducted on 170 specimens of SCCs and AKs (including the ones that underwent metabolomic analysis). SCCs and AK I were found to be significantly associated in terms of the content of some metabolites. Moreover, in the logistic regression model, the presence of parakeratosis in AKs appeared to be less frequently associated with SCCs, while AKs with hypertrophy had a two-fold higher risk of being associated with SCC. Conclusions—Our findings, derived from metabolomics and histopathological data, support the notion that AK I are different from healthy skin and share some different features with SCCs. This may further support the expanding notion that all AKs should be treated independently from their clinical appearance or histological grade because they may be associated with SCC.

2021 - Synergic effect of buccal fat pad pedicled flap and dermal acellular matrix for large cheek defect [Articolo su rivista]
Rossi, E.; Salgarelli, A. C.; Mandel, V. D.; Magnoni, C.
abstract

Reconstruction of large defects of the upper cheek defects still remains a challenge for the surgeon, who can apply different techniques. We present a new method involving the use of a dermal regeneration template to achieve an improved, faster healing of pedicled buccal fat flap in a 75-year-old woman affected by melanoma of the upper-middle cheek. The tumor involved soft tissue, zygomatic arch and periocular fact. The choice of the surgical technique consisted first in the creation of a buccal fat pad to restore the important lack of tissue over the underlying bones, and then in the position of a dermal acellular matrix (Integra® Dermal Regeneration Template; Integra LifeSciences Holdings Corporation, Plainsboro, NJ, USA). Three weeks later, once the neodermal formation was finished, a split thickness graft was placed. This is a not yet described association that represents a good surgical option for the restoration of large cheek defects that allows good functional and cosmetic result in older patient when minimal surgical invasion and operative duration are necessary because of a patient's general condition. The postoperative course with this surgical technique was regular and a good functional result was achieved. This technique provides an adequate functional coverage, a restoration of soft tissue lacking and an acceptable cosmetic result without ectropion.

2020 - Acute maculopapular eruption in Covid-19 patient: A case report [Articolo su rivista]
Rossi, E.; Lasagni, C.; Trakatelli, M.; Wertzberger Rowan, S.; Magnoni, C.
abstract

We report the case of a positive COVID-19 patient who presented to our hospital for a maculopapular skin rash which appeared 7 days after the onset of COVID-19 symptoms. He was 34 years old and nothing relevant was recorded at his previous anamnesis. The patient was hospitalized for 3 days and received systemic therapy with steroid, antihistamines, tocilizumab, and hydroxicloroquine. On the third day of the hospitalization the cutaneous rash had almost completely disappeared.

2020 - An intraoperative study with ex vivo fluorescence confocal microscopy: diagnostic accuracy of the three visualization modalities [Articolo su rivista]
Reggiani, C.; Pellacani, G.; Reggiani Bonetti, L.; Zanelli, G.; Azzoni, P.; Chester, J.; Kaleci, S.; Ferrari, B.; Bellini, P.; Longo, C.; Bertoni, L.; Magnoni, C.
abstract

Ex-vivo fluorescence confocal microscopy (FCM) is an imaging technique that captures real time digital images of freshly excised tissue with quasi histopathological resolution. FCM reduces time to image acquisition compared to hematoxylin and eosin (H&E) frozen section analysis.

2020 - Buschke-Lowenstein tumor: Use of dermal matrix for reconstruction of genital area [Articolo su rivista]
Condorelli, A. G.; Ferrari, B.; Francomano, M.; Rocco, B.; Sighinolfi, M. C.; Giacomelli, L.; Pignatti, M.; Magnoni, C.
abstract

2020 - Clinical Factors Influencing the Outcomes of an Acellular Dermal Matrix for Skin Cancer Treatment: A Retrospective Study [Articolo su rivista]
Ferrari, B.; Reggiani, C.; Francomano, M.; Bellini, P.; Ferrari, F.; Giacomelli, L.; Mannocci, A.; Magnoni, C.
abstract

OBJECTIVE: To evaluate the efficacy and safety of a biologically engineered dermal matrix used in reconstructive surgery after skin tumor resection, focusing on the frequency of successful grafting and identifying potential factors influencing treatment outcomes. DESIGN AND PARTICIPANTS: This retrospective analysis involved consecutive patients diagnosed with skin cancer in any area of the body and for which treatment with a dermal skin template was recommended as alternative to traditional surgery. MAIN OUTCOME MEASURES: Percentage of successful grafting and the patient and tumor characteristics influencing treatment outcome via univariate analysis. MAIN RESULTS: A total of 302 patients were included. Surgical reconstruction with the matrix was effective in 88.9% of the patients within 21 days of surgery. Notably, the matrix was successful regardless of tumor location, type, or size. Infection was the only variable significantly associated with graft failure (P < .001). CONCLUSIONS: The studied dermal matrix provides an efficient alternative to traditional reconstructive surgery in patients who present specific comorbidities or risk factors. The only variable significantly associated with graft failure was infection, which should be properly controlled through appropriate treatment.

2020 - Comparison of two perioperative antibiotic schedules in patients undergoing surgical reconstruction with dermal matrix after excision of skin cancer [Articolo su rivista]
Fiorentini, Chiara; Bedini, Andrea; Mandel, Victor Desmond; Bacca, Erica; Menozzi, Marianna; Reggiani, Camilla; De Pace, Barbara; Meschiari, Marianna; Santoro, Antonella; Franceschini, Erica; Mussini, Cristina; Terrenato, Irene; Giacomelli, Luca; Magnoni, Cristina
abstract

Perioperative antibiotic treatment duration in skin reconstruction with dermal substitutes is not well established. This study compares the incidence of infective complications after two different durations of perioperative antibiotic treatment in patients undergoing surgical reconstruction with skin dermal substitutes (SDS) after excision of skin cancer. Infective complications at the site of SDS were compared in subjects undergoing surgical reconstruction who received either a > 24-hour (extended protocol) or a ≤ 24-hour (short protocol) perioperative antibiotic treatment. Of 116 patients undergoing SDS surgical reconstruction, 62 (53.4%) received an extended schedule, and 54 (46.6%) received a short schedule. The two groups were similar for gender, age, comorbidities, American Society of Anesthesiologists score, and type of skin cancer. Overall incidence rate of infection was 20.7% (24/116). No differences in terms of risk of infection were observed between the two groups (OR: 1.04, 95% CI: 0.42-2.55; P = .937). Patients undergoing SDS reconstruction in the limb/foot had a higher risk of infection in comparison with those undergoing SDS reconstruction in the chest/head (OR: 2.69, 95% CI: 1.06-6.86; P = .038). The short protocol was demonstrated to be beneficial to patients undergoing surgical reconstruction with SDS. A ≤ 24-hour perioperative antibiotic schedule did not increase the infection rate, potentially allowing a reduction of antibiotic exposure.

2020 - Extracellular matrix deposition by adipose-derived stem cells and fibroblasts: a comparative study [Articolo su rivista]
Paganelli, A.; Benassi, L.; Rossi, E.; Magnoni, C.
abstract

Cell-based strategies are today widely studied as possible therapies for wound healing. In this setting, fibroblasts play a key role since they are the main dermal cellular component and are responsible for extracellular matrix secretion. Several works report on the possibility of using fibroblast-derived extracellular matrix scaffolds for wound healing in skin injuries. While fibroblast-based substitutes have already been intensively studied by other groups, we focused our attention on the possibility of creating an adipose-derived stem cell (ADSC)-induced dermal scaffold for wound healing. ADSCs are a particular subset of mesenchymal stem cells present in the stromal vascular fraction of the adipose tissue. The aim of our work was to compare the ability of ADSCs and fibroblast to produce in vitro a scaffolding material, both in terms of collagen and fibronectin production. ADSCs turned out to be capable of efficiently producing a collagen and fibronectin-containing dermal matrix upon stimulation with ascorbic acid. We observed fibronectin and collagen production by ADSCs to be even more abundant when compared to fibroblasts’. Our results support the use of ADSC-induced sheets instead of fibroblast-based dermal substitutes as wound-healing strategies in full-thickness skin injuries.

2020 - Hidradenitis suppurativa: Surgical and postsurgical management [Articolo su rivista]
Manfredini, M.; Garbarino, F.; Bigi, L.; Pellacani, G.; Magnoni, C.
abstract

Background: Hidradenitis suppurativa (HS) is a chronic inflammatory disorder. Several medical treatments, with varying degrees of efficacy, have been developed. However, in most cases of advanced HS, the definitive treatment option is often represented by surgical excisions. Objective: Surgical techniques, reconstructive approach, and local wound care should be accurately designed in order to obtain the best result. In this review we analyze the possible surgical treatments and local wound care. Methods: A MEDLINE search was performed on the various surgical treatments, reconstructive techniques, and local wound care. Results: Surgical treatment is a common therapeutic modality for HS. Different surgical reconstructive techniques and post-surgical wound care approaches are described for the management of HS patients. Conclusions: There were few high-quality evidence-based studies evaluating the surgical management of HS. Many disparate HS severity scores were used in these studies, making comparisons between them difficult. Nonetheless, research on different surgical approaches and wound care management has increased substantially in the past decade and it has given patients more surgical therapeutic strategies. The description of the best combinations and timing of surgery, wound care, and medical therapies will be a matter of future research for the definition of the optimal management of the HS patient.

2020 - Histopathological determinants of autofluorescence patterns in oral carcinoma [Articolo su rivista]
Meleti, M.; Giovannacci, I.; Vescovi, P.; Pedrazzi, G.; Govoni, P.; Magnoni, C.
abstract

Biological tissues (including oral mucosa) can absorb and re-emit specific light wavelengths, detectable through spectrophotometric devices. Such a phenomenon is known as “autofluorescence” (AF). Several devices evaluating tissue AF have been developed and commercialized in the last two decades. Among these, the VELscope® system has been proposed as a visual diagnostic aid for potentially malignant disorders and malignant lesions of the oral mucosa. In the present pilot study, we investigated which are the main histopathological features possibly related to variations in AF patterns in a set of 20 oral squamous cell verrucous carcinoma. Among all the histological features investigated, only the mean width of keratin was significantly different between hypofluorescent and hyperfluorescent carcinomas. The results of the present study demonstrate that AF features of oral malignant lesions are significantly associated with the width of their keratin layer.

2020 - Mesenchymal stem cells and psoriasis: State of the art and future perspectives [Articolo su rivista]
Paganelli, A.; Kaleci, S.; Benassi, L.; Pellacani, G.; Magnoni, C.
abstract

2020 - Mesenchymal stem cells for the treatment of psoriasis: a comprehensive review [Articolo su rivista]
Paganelli, A.; Tarentini, E.; Benassi, L.; Kaleci, S.; Magnoni, C.
abstract

Mesenchymal stem cells (MSCs) have recently been shown to have not only regenerative capabilities but also immunomodulating properties. For this reason, they are currently under investigation in clinical trials for the treatment of several autoimmune systemic disorders. Psoriasis is a systemic immune-mediated disease for which MSCs could have therapeutic potential. We analysed the existing literature with regard to MSC-based strategies for the treatment of psoriasis, using the MEDLINE, Embase, Scopus and Cochrane Library electronic databases from inception to the date of study. A number of studies confirm the involvement of MSCs in psoriasis pathogenesis and therefore designate MSCs as an important potential therapeutic tool in this setting. Preclinical data are mostly based on imiquimod-induced murine models of psoriasis, and confirm the anti-inflammatory and immunomodulatory action of MSCs in the setting of psoriasis. Six patients affected by psoriasis were described in four clinical studies. Despite significant differences in terms of therapeutic protocols and clinical outcomes, the MSC-based regimens were efficacious in 100% of the cases. Despite more data still being needed, MSCs could be a promising therapy for psoriasis.

2020 - Salivary microRNA for diagnosis of cancer and systemic diseases: A systematic review [Articolo su rivista]
Setti, G.; Pezzi, M. E.; Viani, M. V.; Pertinhez, T. A.; Cassi, D.; Magnoni, C.; Bellini, P.; Musolino, A.; Vescovi, P.; Meleti, M.
abstract

Background: The aberrant expression of microRNAs (miRNAs) has been associated with several diseases, including cancer, inflammatory, and autoimmune conditions. Interest in salivary miRNAs as non-invasive tools for the diagnosis of malignancies and systemic diseases is rapidly increasing. The present systematic review was developed for answering the question: “Are salivary microRNAs reliable biomarkers for diagnosis of cancer and systemic diseases?” Methods: The application of inclusion and exclusion criteria led to the selection of 11 papers. Critical appraisals and quality assessments of the selected studies were performed through the National Institute of Health “Study Quality Assessment Tool” and the classification of the Oxford Center for Evidence-Based Medicine. Results: Seven studies reported statistically significant correlations between one or more salivary miRNAs and the investigated disease. The critical analysis allowed us to classify only two studies (18.2%) as having “good” quality, the rest being scored as “intermediate” (8; 73%) and “poor” (1; 9%). Evidence exists that salivary miR-940 and miR-3679-5p are reliable markers for pancreatic cancer and that miR140-5p and miR301a are promising molecules for the salivary diagnosis of gastric cancer. Conclusions: Further studies, possibly avoiding the risk of bias highlighted here, are necessary to consolidate these findings and to identify new reliable salivary biomarkers.

2020 - Surgical and postsurgical wound care in hidradenitis suppurativa [Articolo su rivista]
Manfredini, M.; Garbarino, F.; Bigi, L.; Pellacani, G.; Magnoni, C.
abstract

Hidradenitis suppurativa (HS) is a chronic inflammatory disorder. Several medical treatments, with varying degree of efficacy, have been developed. However, in most cases of advanced (HS), the definitive treatment option is often represented by surgical excisions. Surgical techniques, reconstructive approach, and local wound care should be accurately designed in order to obtain the best result. In this letter, we analyzed the possible surgical treatments and local wound care. A literature review was performed on the various surgical treatments, reconstructive techniques, and local wound care. Surgical treatment is a common therapeutic modality for HS. Different surgical reconstructive techniques and postsurgical wound care approaches are described for the management of HS patients. There were few high-quality evidence-based studies evaluating the surgical management of HS. Many disparate HS severity scores were used in these studies making comparison between them difficult. Nonetheless, research into different surgical approaches and wound care management has increased substantially in the past decade and has given patients more surgical therapeutic strategies. The description of the best combinations and timing of surgery, wound care and medical therapies, will be a matter of future research for the definition of the optimal management of HS patient.

2020 - Surgical management of squamous cell carcinoma arising in patients affected by epidermolysis bullosa: a comparative study [Articolo su rivista]
Paganelli, A.; Reggiani, C.; Fiorentini, C.; Lando, M.; Cesinaro, A. M.; Magnoni, C.
abstract

2020 - The COVID-19 outbreak in dermatologic surgery: resetting clinical priorities [Articolo su rivista]
Rossi, E.; Trakatelli, M.; Giacomelli, L.; Ferrari, B.; Francomano, M.; Pellacani, G.; Magnoni, C.
abstract

Emilia-Romagna was one of the Italian regions mostly affected by the COVID-19 pandemic and lockdown measures were taken to slow the COVID-19 outbreak. All routine activities in Modena hospitals were suspended; however, urgent procedures were still to be performed. Setting the priority of procedures in oncological dermatology in the COVID-19 era is challenging.

2019 - Auto-Fluorescence and Histopathologic Evaluation of Medication-Related Osteonecrosis of the Jaws: Perspectives for Treatment [Articolo su rivista]
Giovannacci, Ilaria; Vescovi, Paolo; Magnoni, Cristina; Corradi, Domenico; Corcione, Luigi; Lucchina, Alberta Greco; Mortellaro, Carmen; Nammour, Samir; Meleti, Marco
abstract

Auto-fluorescence (AF) of healthy bone tissue has recently been described. Loss of AF (LAF) has, on the contrary, been reported in necrotic bone. Further, the use of LAF as a possible guidance to distinguish viable from necrotic bone during surgical treatment of osteonecrosis has been proposed. The aim of this study is to detail 8 patients of medication-related osteonecrosis of the jaws treated through an AF-guided surgical resection. The authors also provide the histopathologic description of hypo-fluorescent and hyper-fluorescent bone in each patient. After removal of necrotic bone block, Er:YAG laser was used for vaporizing further necrotic bone, up to the detection of strongly hyper-fluorescent bone. Samples of hyper-fluorescent bone were collected around areas of necrosis. Histopathologic evaluation revealed viable bone tissue in all hyper-fluorescent specimens. On the basis of these data, AF-guided surgical resection could be effective in highlighting surgical margins of necrotic bone tissue and it might have some utility in a range of applications of bone surgery.

2019 - Effects of topical methotrexate loaded gold nanoparticle in cutaneous inflammatory mouse model [Articolo su rivista]
Fratoddi, I.; Benassi, L.; Botti, Eleonora; Vaschieri, C.; Venditti, I.; Bessar, H.; Samir, M. A.; Azzoni, P.; Magnoni, C.; Costanzo, A.; Casagrande, V.; Federici, M.; Bianchi, L.; Pellacani, G.
abstract

Gold nanoparticles functionalized with 3-mercapto-1 -propansulfonate (AuNPs-3MPS) have been prepared and loaded with Methotrexate (MTX), an immunosuppressive agent used in the systemic treatment of moderate-severe inflammatory diseases. The effects of the AuNPs-3MPS@MTX topically administered in vitro on skin model and in vivo on imiquimod-induced psoriasis-like mice model, have been studied. Clinical response, epidermal thickness, cell proliferation rate and inflammation were tested. AuNPs-3MPS@MTX treated mice showed a decreasing of scaling and erythema score, reduction of epidermal thickness, parakeratosis and hyperkeratosis, compared to AuNPs3MPS treated mice. Immunnhistocheinistry analysis staining displayed that Ki67, K6 CD3 and CD8 stainings were reduced in AuNPs3MPS@MTX treated mice. Blood evaluation showed no differences in blood count and in ALT and AST levels before and after AuNPs3MPS or AuNPs-3MPS@MTX treatment. Topical AuNPs-3MPS@MTX treatment is able to induce a reduction of keratinocytes hyperproliferation, epidermal thickness and also inflammatory infiltrate in vivo on imiquimod-induced psoriasis like mice model. Published by Elsevier Inc.

2019 - Field cancerization therapy with ingenol mebutate contributes to restoring skin-metabolism to normal-state in patients with actinic keratosis: a metabolomic analysis [Articolo su rivista]
Righi, V.; Tarentini, E.; Mucci, A.; Reggiani, C.; Rossi, M. C.; Ferrari, F.; Casari, A.; Magnoni, C.
abstract

Actinic keratosis (AK) is a skin premalignant lesion, which progresses into squamous cell carcinoma (SCC) if left untreated. Ingenol mebutate gel is approved for local treatment of non-hyperkeratotic, non-hypertrophic AK; it also has the potential to act as a field cancerization therapy to prevent the progression of AK to SCC. To gain better insights into the mechanisms of ingenol mebutate beyond the mere clinical assessment, we investigated, for the first time, the metabolome of skin tissues from patients with AK, before and after ingenol mebutate treatment, with high-resolution magic angle spinning nuclear magnetic resonance spectroscopy. The metabolomic profiles were compared with those of tissues from healthy volunteers. Overall, we identified a number of metabolites, the homeostasis of which became altered during the process of tumorigenesis from healthy skin to AK, and was restored, at least partially, by ingenol mebutate therapy. These metabolites may help to attain a better understanding of keratinocyte metabolism and to unmask the metabolic pathways related to cell proliferation. These results provide helpful information to identify biomarkers with prognostic and therapeutic significance in AK, and suggest that field cancerization therapy with ingenol mebutate may contribute to restore skin metabolism to a normal state in patients with AK.

2019 - From actinic keratosis to squamous cell carcinoma: NMR analysis with clinical and histological aspects [Abstract in Atti di Convegno]
Righi, Valeria; Tarentini, Elisabetta; Rossi, Maria Cecilia; Magnoni, Cristina; Mucci, Adele
abstract

2019 - In vitro Engineering of a Skin Substitute Based on Adipose-Derived Stem Cells [Articolo su rivista]
Paganelli, A.; Benassi, L.; Pastar, I.; Pellegrini, M.; Azzoni, P.; Vaschieri, C.; Pisciotta, A.; Carnevale, G.; Pellacani, G.; Magnoni, C.
abstract

In the field of wound healing, stem cell-based strategies are gaining importance for their regenerative potential. Adipose-derived stem cells (ADSCs) are a particular subset of mesenchymal stem cells present in the stromal-vascular fraction of the adipose tissue, today considered very attractive for their relative abundance and accessibility in the human body. However, ADSCs are still not routinely used in normal clinical practice. Several studies have also reported ADSC transplantation in association with biomaterials in an attempt to enhance the local retention and growth rate of the cells. The aim of our study was to evaluate the ability of ADSCs to build a dermal scaffold to be potentially used as a dermal substitute in the field of wound healing, with optimal biocompatibility and mechanical properties. ADSCs were defined as CD90-, CD73-, and CD105-positive cells. ADSCs turned out to be capable of secreting all the main components of the extracellular matrix (ECM) upon stimulation, thus efficiently producing a collagen and fibronectin-containing dermal matrix. We also checked whether the ADSC-produced dermal scaffold could be seeded with keratinocytes. The scaffolding material directly produced by ADSCs has several advantages when compared to the commercially available ones: it is easily obtained from the patients and it is 100% biocompatible and supports cell-ECM interaction. Moreover, it represents a possible powerful therapeutic tool for patients with chronic ulcers since it appears to be potentially grafted with keratinocytes layers, thus bypassing the classical two-step grafting procedure.

2019 - Integra in Scalp Reconstruction After Tumor Excision: Recommendations From a Multidisciplinary Advisory Board [Articolo su rivista]
Magnoni, Cristina; De Santis, Giorgio; Fraccalvieri, Marco; Bellini, Pietrantonio; Portincasa, Aurelio; Giacomelli, Luca; Papa, Giovanni
abstract

Integra is a dermal regeneration template used in the reconstruction of burns, traumatic injuries, or excision lesions in patients who present particular risk factors for traditional surgical procedures. A multidisciplinary advisory board of expert dermatologists and plastic surgeons have discussed the use of Integra in the reconstruction of scalp defects after tumor excision, focusing on the evidence derived from literature and on their experience in the treatment of approximately 400 patients. In this position paper, the authors summarize the main evidence discussed during the board, and the common practice guidelines proposed by the experts. The use of Integra is recommended in elderly patients with multiple comorbidities who have a higher risk for potential complications in traditional surgery; these patients may in fact benefit from a lower anesthetic risk, a less complicated postsurgical care and limited morbidity at the donor site obtained with the dermal template. Integra should also be used in the reconstruction of large and complex wounds and in case of bone exposure, as it helps to overcome the challenges related to wound healing in difficult areas. Notably, Integra has proven to be effective in patients who have undergone previous surgical procedures or adjuvant radiation therapy, in which previous incisions, scarring and radiation damages may hamper the effectiveness of traditional procedures. Finally, Integra is recommended in patients with recurrent and aggressive tumors who need closer tumor surveillance, as it gives easy access to the tumor site for oncologic follow-up examination.

2019 - Salivary 1H-NMR Metabolomics in Primary Sjögren Syndrome. Preliminary Results of a Pilot Case-Control Study [Abstract in Rivista]
Setti, G.; Sandri, G.; Tarentini, E.; Panari, L.; Mucci, A.; Righi, V.; Meleti, M.; Magnoni, C.; Consolo, U.; Bellini, P.
abstract

2019 - Self-healing CD30- T-clonal proliferation of the tongue: report of an extremely rare case [Articolo su rivista]
Setti, Giacomo; Martella, Eugenia; Mancini, Cristina; Vescovi, Paolo; Magnoni, Cristina; Bellini, Pierantonio; Giovannacci, Ilaria; Meleti, Marco
abstract

The etiology of traumatic ulcerative granulomas with stromal eosinophilia (TUGSE) is not clear, traumatic irritation having advocated as the most likely cause. TUGSEs are typically self-limiting slow-healing lesions of the oral mucosa with unclear pathogenesis, commonly manifesting as a rapidly developing, long-lasting ulcer.

2019 - The “Bat Flap”: A flap for reconstruction of large skull skin defects [Articolo su rivista]
Reggiani, C.; Bellini, P.; Salgarelli, A. C.; Setti, G.; Ferrari, B.; De Santis, G.; Starnoni, M.; Magnoni, C.
abstract

2019 - Two-step surgery in extensive anogenital hidradenitis: What kind of faecal management? [Articolo su rivista]
Garbarino, F.; Manfredini, M.; Rossi, E.; Magnoni, C.
abstract

2019 - Which are the main fluorophores in skin and oral mucosa? A review with emphasis on clinical applications of tissue autofluorescence [Articolo su rivista]
Giovannacci, I.; Magnoni, C.; Vescovi, P.; Painelli, A.; Tarentini, E.; Meleti, M.
abstract

Abstract OBJECTIVES: The present review provides information about which molecules appear to be the main fluorophores in skin and oral mucosa, together with their clinical applications. DESIGN: The MEDLINE database was searched, using "oral mucosa AND fluorophores", "skin AND fluorophores", "epidermal AND fluorophores", "dermal AND fluorophores" and "cutaneous AND fluorophores" as entry terms. We searched the literature following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The level of evidence in the studies was assessed using the Classification of the Oxford Centre for Evidence-based Medicine (CEBM) Levels for Diagnosis. RESULTS: Five papers and 17 were primarily focused on description of fluorophores in oral mucosa and skin Evidence exists that fluorophores of oral mucosa and skin are mainly proteins such as collagen, elastin, keratin and tryptophan. Other possible fluorophores identified are: porphyrins, advanced glycation end products, flavins, lipopigment, nicotinamide adenine dinucleotide, flavin adenine dinucleotide, pheomelanin, eumelanin and components of lipofuscin. Clinical applications of oral mucosal autofluorescence (AF) are related to management of malignant and potentially malignant lesions. In the skin, AF has been used for acne assessment, diagnosis of sweat-gland pathologies, glycemic control and management of malignant lesions and as a marker for skin aging. CONCLUSION: Fluorophores stimulated through AF devices are implied in different physiologic and pathologic processes. AF seems to be useful for several clinical applications, especially in skin department. Because most of the studies show a low level of evidence, further studies are necessary in such a promising and fascinating field.

2018 - Identifying the factors that influence surgeon's compliance with excisional margins of non-melanoma skin cancer [Articolo su rivista]
Cautela, Jennifer Marchetti; Mannocci, Alice; Reggiani, Camilla; Persechino, Flavia; Ferrari, Federica; Rossi, Elena; Passini, Erika; Bellini, Pierantonio; Meleti, Marco; WERTZBERGER ROWAN, Sara; Magnoni, Cristina
abstract

The rising incidence of Non Melanoma Skin Cancers (NMSC) leads to a high number of surgical procedures worldwide. The strict compliance with international guidelines with regard to excisional margins may help decrease the number of re-excision procedures and reduce the risk of NMSC recurrence. The aim of this study was to investigate the prevalence of excisional margins as recommended by the European Academy of Dermatology and Venereology (EADV) and the European Dermatology Forum (EDF) guidelines, and the factors (demographic or clinical) that influence surgeons' compliance with these guidelines.This was a prevalence study looking at surgical excisions of NMSCs performed over a period of 2 years (2011-2012). A sample size of 1669 patients was considered. Definition of excisional margins recommended by the international guidelines (EADV and EDF) were used as point of reference for the analysis. Tumor and histologic specimen size were calculated ex vivo by 5 different pathologists. The size of skin specimens was measured with a major axis and a minor axis. The same was done for the tumor present on the skin specimens. The differences between the major and minor axes of surgical specimen and tumor were calculated. These differences were subsequently divided by two, hypothesizing that the lesion had the same distance from the margins of the surgical specimen. The differences obtained were named "Delta", the formulas applied being the following: Delta major = (major axis specimen-major axis tumor)/2; Delta minor = (minor axis specimen -minor axis tumor)/2.Results show a significant statistical difference, associated with factors such as: Age of the patient, anatomical localization of the tumor, histological diagnosis, and surgeons' experience.The identification of these factors sheds light on clinicians' practice and decision-making regarding excisional margins. Hopefully a higher level of adherence to the guidelines can be achieved in the future.

2018 - Infective complications in patients undergoing surgical reconstruction with dermal matrix: the Modena experience [Poster]
Bedini, Andrea; Fiorentini, Chiara; Mandel, Victor Desmond; Menozzi, Marianna; Bacca, Erica; Ferrari, Federica; DE PACE, Barbara; Meschiari, Marianna; Santoro, Antonella; Orlando, Gabriella; Franceschini, Erica; Puzzolante, Cinzia; Mussini, Cristina; Magnoni, Cristina
abstract

Background: Bioengineered skin dermal substitutes (SDS) represent a novel therapeutic opportunity for restoring damaged tissue1,2,3. Antimicrobial prophylaxis duration in such procedures has not been well established yet. The aim of the study was to evaluate the changing of infective complications following shortening of perioperative prophylaxis in patients undergoing surgical reconstruction with SDS. Material & Methods: Infective complications at the site of SDS were compared in 2 groups: subjects undergoing surgical reconstruction between September 2014 and January 2016 (PERIOD A) who received a >24H-antibiotic-prophylaxis, and between May 2016 and June 2017 (PERIOD B) who received a ≤24H-antibiotic-prophylaxis. Differences in the incidence of infection and pathogen prevalence were explored. Results: Between September 2014 and June 2017, 116 patients underwent a surgical reconstruction with a SDS. The mean age was 73-years, 77 were male (66.4%), 78 (67.2%) were positive for hypertension, 20 (17.2%) for diabetes mellitus, 16 (13.8%) for chronic renal impairment, 22 (19%) were smokers, and 45 (38.8%) had an ASA score ≥3. In the 94.8% (n=110) the reason of surgical intervention was a skin cancer. Surgical SDS reconstruction involved the scalp in 44 cases (37.9%), the face in 28 (24.1%), the chest in 11 (9.5%), the limbs in 33 (28.5%). Among 116 patients, 62 (53.4%) received >24H-antibiotic and 54 (46.6%) ≤24H-antibiotic-prophylaxis. The average duration of prophylaxis in the 2 groups of patients was 6.6 days and 0.5 day, respectively. Overall incidence rate of infection was 20.7% (24/116). The most frequently isolated pathogen was S. aureus (41.6%), followed by P. aeruginosa (29.1%), P. mirabilis (8.3%), and E. faecalis (4.1%). Patients undergoing SDS reconstruction in limbs had higher infection rate in comparison with chest/head (33.3% and 15.6%, respectively; p=0.034). No differences in the infection rate were observed between the patients who received >24H or ≤24H-antibiotic-prophylaxis (22.5% and 18.5%, respectively; p=0.590). The two groups resulted similar for gender, age, comorbidities, ASA score, and type of skin cancer. Discussion: As far as we know, this is the first study that compared two perioperative antibiotic prophylaxis regimes in patients undergoing SDS reconstruction. Comparing the two patient groups (≤24-hour and >24-hour prophylaxis), no differences in the rate of infection were found. The result is very important: it shows that prolongation of prophylaxis in this type of surgical patients does not reduce the rate of infection. Shortening of antibiotic prophylaxis allowed to reduce of 6 days-per-patient the antibiotic exposure. It was surprising that only the reconstruction of the limbs, in comparison with other sites, was associated with a higher risk of infection (33.3 and 15.7 respectively). Nor the most critical patients (with an ASA score ≥3), nor patients undergoing major surgical reconstructions (surgical area >60 cm2) resulted associated with a higher risk. Conclusion: Antibiotic prophylaxis reduction to 24 hours or less demonstrated to be beneficial to patients undergoing surgical reconstruction with SDS. Shortening of antibiotic prophylaxis did not increase infection rate, and it allowed a reduction of 6 days-per-patient the antibiotic exposure. Randomized and controlled trials, with greater population, could give a more accurate response on the duration of antibiotic prophylaxis in patients undergoing surgical SDS reconstruction.

2018 - Oral melanoma and other pigmentations: when to biopsy? [Articolo su rivista]
Lambertini, M; DI PATRIZI, Alfonso; Fanti, P A; Melotti, B; Caliceti, U; Magnoni, C; Misciali, C; Baraldi, C; Ravaioli, G M; Dika, E
abstract

Oral pigmentations (OPs) are often neglected, although a meticulous examination of the oral cavity is important not only in the diagnosis of oral melanoma, but also for the detection of important clinical findings that may indicate the presence of a systemic disease. OPs may be classified into two major groups on the basis of their clinical appearance: focal and diffuse pigmentations, even though this distinction may not appear so limpid in some cases. The former include amalgam tattoo, melanocytic nevi, melanoacanthoma and melanosis, while the latter include physiological/racial pigmentations, smoker's melanosis, drug-induced hyperpigmentations, postinflammatory hyperpigmentations and OPs associated with systemic diseases. We will discuss the most frequent OPs and the differential diagnosis with oral mucosal melanoma (OMM), underlining the most frequent lesions that need to undergo a bioptic examination and lesions that could be proposed for a sequential follow-up.

2018 - The smart approach: feasibility of lentigo maligna superficial margin assessment with hand-held reflectance confocal microscopy technology [Articolo su rivista]
Pellacani, G.; De Carvalho, N.; Ciardo, S.; Ferrari, Beatrice; Cesinaro, A. M.; Farnetani, F.; Bassoli, S.; Guitera, P.; Star, P.; Rawson, R.; Rossi, E.; Magnoni, C.; Gualdi, G.; Longo, C.; Scope, A.
abstract

Background: Lentigo maligna may be challenging to clear surgically. Objective: To evaluate feasibility of using superficial skin cuts as RCM imaging anchors for attaining negative surgical margins in lentigo maligna. Methods: Included patients presented with lentigo maligna near cosmetically sensitive facial structures. We evaluated, with hand-held-RCM, microscopic clearance of melanoma beyond its dermoscopically detected edges. Evaluated margins were annotated using shallow skin cuts. If a margin was positive at ‘first-step’ RCM evaluation, we sequentially advanced the margin radially outward at that segment by 2-mm intervals until an RCM-negative margin was identified. Prior to final surgical excision, we placed sutures at the outmost skin cuts to allow comparison of RCM and histopathological margin assessments. Primary outcome measure was histopathological verification that RCM-negative margins were clear of melanoma. Results: The study included 126 first-step margin evaluations in 23 patients, median age 70 years (range: 43–91). Seventeen patients (74%) had primary in-situ melanoma and six (26%) invasive melanoma, mean thickness 0.3 mm (range 0.2–0.4 mm). Six cases (26%) showed complete negative RCM margins on ‘first-step’, 11 (48%) were negative at ‘second-step’, and four (17%) at ‘third-step’. In two additional cases (9%), margins clearance could not be determined via RCM due to widespread dendritic cells proliferation. The RCM-negative margins in all 21 cases proved clear of melanoma on histopathology. Of the 15 cases that returned at 1-year follow-up, none showed any residual melanoma on dermoscopic and RCM examinations. Interobserver reproducibility showed fair agreement between bedside RCM reader and blinded remote-site reader, with Spearman's rho of 0.48 and Cohen's kappa of 0.43; using bedside reader as reference, the remote reader's sensitivity was 92% and specificity 57% in positive margin detection. Conclusions: Margin mapping of lentigo maligna with hand-held-RCM, using superficial skin cuts, appears feasible. This approach needs validation by larger studies.

2018 - Treatment of chronic infected post-oncological wounds with a dermal matrix: two case studies [Articolo su rivista]
Mandel, Victor Desmond; Persechino, Flavia; Fiorentini, Chiara; Passini, Erica; Magnoni, Cristina
abstract

The reconstruction of complex wounds in patients with comorbidities in the lower extremities is a challenging problem for surgeons. Skin grafting is frequently used to cover large skin defects, but it has several limits, including unwanted outcomes resulting from scars, poor elasticity and limitations in joint movement due to contractures. Locoregional flaps, particularly in the lower limbs, have limited application due to the size of the defect. Tissue engineering of the skin has offered major improvements in the coverage of large defects. Dermal matrix can be applied in order to generate a new dermis that allows good re-epithelialisation or skin grafting at a later stage. The reconstruction of large lower limb defects is more complicated in the case of chronic wounds showing no tendency to heal due to chronic infection. For all surgeons, it is very important to prevent the formation of a biofilm or manage it when it is already established before undertaking surgical procedures that involve a dermal matrix. We report our reconstruction strategy of chronic infected neoplastic ulcers of the lower extremities with a dermal matrix and our postoperative dressing protocol.

2017 - Case of bullous pemphigoid in a 28-year-old woman affected by tuberous sclerosis complex [Articolo su rivista]
Mandel, Victor Desmond; Fiorentini, Chiara; Benati, Elisa; Benassi, Luisa; Magnoni, Cistina; Pellacani, Giovanni
abstract

[No abstract available]

2017 - Changing in the post-surgery infective complications following the shortening of the antibiotic prophylaxis in the patients undergoing skin dermal substitutes reconstruction [Poster]
Bedini, Andrea; Fiorentini, Chiara; Mandel, Victor Desmond; Bacca, Erica; Ferrari, Federica; DE PACE, Barbara; Meschiari, Marianna; Santoro, Antonella; Orlando, Gabriella; Franceschini, Erica; Puzzolante, Cinzia; Mussini, Cristina; Magnoni, Cristina
abstract

Background: Bioengineered skin dermal substitutes (SDS) represent a novel therapeutic opportunity for restoring damaged tissue, both in massive deep burns, extensive full-thickness wounds, and reconstruction after cancer resection. Antimicrobial prophylaxis duration in such procedures has not been well established yet. The aim of the study was to evaluate the changing of infective complications following shortening of perioperative prophylaxis in patients undergoing surgical reconstruction with SDS. Material & Methods: Infective complications at the site of SDS were compared in two groups of patients: subjects undergoing surgical reconstruction between September 2014 and January 2016 (PERIOD A) who received a >24h-antibiotic prophylaxis, and subjects undergoing surgical reconstruction between May 2016 and June 2017 (PERIOD B) who received a ≤24h-antibiotic prophylaxis. Differences in the incidence of infection and pathogen prevalence were explored. Univariate linear regression analysis was performed to evaluate the risk factors for infection (sex, age, ASA code, perioperative antibiotic prophylaxis, site of SDS intervention, type of SDS, dimensions of surgical area, chronic renal impairment, and diabetes mellitus). Results: Between September 2014 and June 2017, 116 patients underwent a surgical reconstruction with a SDS. The 66.4% (n=77) of the study population was male, and the mean age was 73 years (22-92 years). Seventy-eight patients (67.2%) were positive for hypertension, 20 (17.2%) for diabetes mellitus, 16 (13.8%) for chronic renal impairment, 22 (19%) were former or current smokers, and 45 (38.8%) had an ASA code ≥3. In the 94.8% of the patients (n=110) the reason of surgical intervention was a skin cancer. Surgical SDS reconstruction involved the scalp in 44 cases (37.9%), the face in 28 (24.1%), the chest in 11 (9.5%), the arm or the hand in 9 (7.8%), the leg in 12 (10.3%) and the foot in 12 (10.3%). Among 116 patients undergoing SDS surgical reconstruction, 62 (53.4%) received a >24h-prophylaxis and 54 (46.6%) received a ≤24h-prophylaxis. The average duration of prophylaxis in the 2 groups of patients was 6.6 days and 0.5 day, respectively. Overall incidence rate of infection was 20.7% (24/116). The most frequently isolated pathogen was S. aureus (41.6%), followed by P. aeruginosa (29.1%), P. mirabilis (8.3%), and E. faecalis (4.1%). Patients undergoing SDS reconstruction in limb/foot had higher infection rate in comparison with those undergoing SDS reconstruction in chest/head (33.3% and 15.6%, respectively; p=0.034). No differences in the infection rate were observed between the patients who received >24h or ≤24h-antibiotic prophylaxis (22.5% and 18.5%, respectively; p=0.590). The two groups resulted similar for gender, age, comorbidities, ASA score, and type of skin cancer. No significant differences in pathogen prevalence were found (p=0.692). Conclusion: Antibiotic prophylaxis reduction to 24 hours or less demonstrated to be beneficial to patients undergoing surgical reconstruction with SDS. Shortening of antibiotic prophylaxis did not increase infection rate, and it allowed to reduce of 6 days-per-patient the antibiotic exposure.

2017 - Comprehension of metabolic disorders in inflammatory and neoplastic hyper-proliferative diseases: a NMR contribution [Abstract in Atti di Convegno]
Righi, 112 V.; Mucci, A.; Tarentini, Elisabetta; Magnoni, C.
abstract

A progression model based on genetic alterations has been proposed for several types of tumors and in skin diseases, the concept of cancerization field involves cluster of genetically altered cells in a chronically photo damaged skin without clinical evidence of neoplastic lesions [1]. Psoriasis is a chronic, inflammatory skin disease associated with significant morbidity and mortality. Cutaneous Squamous Cell Carcinoma (CSCC) is a common skin cancer characterized by malignant proliferation of keratinocytes. CSCC usually arises from precursor lesions such as actinic keratosis (AKs), but can also grow de novo or on chronically inflamed skin. AKs are the most common skin lesion of disordered keratinocyte proliferation in the disease continuum of photo-damaged skin that may lead to invasive CSCC. In addition, patients with psoriasis can be developed AK or CSCC. Exploring the metabolome of cancer, precancerous lesions and inflammatory disease seems to be a parallel and effective way to understand the phenotypic changes associated with cancer progression. Metabolomics could reveal novel cancer biomarkers that might expand our current understanding of the multifactorial disease. Nuclear Magnetic Resonance (NMR) spectroscopy, giving an accurate description of the molecular composition of a human tissue, provides a “fingerprint” of the whole metabolome [2]. Correlations between some metabolites and proliferative markers allow gaining insight into the relationship between cellular proliferation and metabolic changes associated with the presence of tumor and its aggressiveness.

2017 - Non-melanoma skin cancer of the head and neck: the aid of reflectance confocal microscopy for the accurate diagnosis and management [Articolo su rivista]
Ferrari, Barbara; Salgarelli, Attilio Carlo; Mandel, Victor Desmond; Bellini, Pierantonio; Reggiani, Camilla; Farnetani, Francesca; Pellacani, Giovanni; Magnoni, Cristina
abstract

Background: Non-melanoma skin cancer (NMSC) represents the most common cutaneous neoplasms of the head and neck. In recent years, novel non-invasive diagnostic tool have been developed, and among these we have the reflectance confocal microscopy (RCM), that offers the evaluation of the skin at real time with cellular resolution. Numerous studies have identified the main confocal features of skin tumours, demonstrating the good correlation of these features with certain dermatoscopic patterns and histologic findings. Evidence acquisition: The aim of this analysis was to provide new insight into the role of RCM in the diagnosis and management of NMSC of the head and neck. Data comes from the most recent literature, taking into account previous essential reported information in this field. The study eligibility criteria were: studies providing update information, focusing on RCM findings in NMSC, without restrictions for age, sex, ethnicity. A search concerning the role of dermoscopy and RCM in the diagnosis of NMSC was performed on Medline. Duplicated studies, single case report and papers with language other than English were excluded from this study. Evidence synthesis: RCM clues were analysed for NMSC in association with clinical, dermoscopic and histopathologic findings. Moreover, some new findings have been described and possible applications for NMSC of the head and neck have been discussed. Conclusions: RCM allows tissue imaging in vivo contributing to a more accurate diagnosis of NMSC of the head and neck, sparing time for the patient and costs for the public health system. RCM can also be used for selection of the biopsy site and it is helpful in defining the surgical safety margins to keep during the excision of skin cancers.

2017 - Old but Gold: The Importance of Medical History in Diagnosing Neutrophilic Dermatoses Characterized by Pathergy. A Case of Pyoderma Gangrenosum [Articolo su rivista]
Paganelli, Alessia; Garbarino, Federico; Fiorentini, Chiara; Bigi, Laura; Magnoni, Cristina
abstract

2017 - Structure-Activity Relationship within a new series of σ1 and σ2 receptor ligands: identification of a novel σ2R agonist (BS148) with selective toxicity against metastatic melanoma [Articolo su rivista]
Franchini, Silvia; Sorbi, Claudia; Battisti, Umberto Maria; Tait, Annalisa; Bencheva, Leda Ivanova; Cichero, Elena; Fossa, Paola; Cilia, Antonio; Prezzavento, Orazio; Ronsisvalle, Simone; Aricò, Giuseppina; Benassi, Luisa; Vaschieri, Cristina; Azzoni, Paola; Magnoni, Cristina; Brasili, Livio
abstract

A new series of spiro-cyclic sigma ligands were prepared and studied. Most were found to have a high affinity and selectivity for σ1R; compounds 7b, 15b and 16a were shown to be σ1R agonists, while 16b was proven to be the only σ1R antagonist. Only 16a (BS148) exhibited σ2R selectivity and was able to inhibit the growth of metastatic malignant melanoma cell lines, whilst not affecting normal human melanocytes. BS148’s anti-proliferative activity suggests a σ2R agonist profile. Further, preliminary investigations indicate that, at least a part, the mechanism of metastatic malignant melanoma cell death induced by BS148 is due to apoptosis.

2017 - Synergic effect of buccal fat pad pedicled flap and dermal acellular matrix for large cheek defect [Poster]
Rossi, Elena; Salgarelli, Attilio Carlo; Mandel, Victor Desmond; Magnoni, Cristina
abstract

Introduction & Objectives: Reconstruction of large defects of the upper cheek defects still remains a challenge for the surgeon, than can apply different techniques. We present a new method involving the use of a dermal regeneration template to achieve an improved, faster healing of pedicled buccal fat flap in a 75-years-old woman affected by melanoma of the upper-middle cheek. The tumor involved soft tissue, zygomatic arch and periocular fact. Material & Methods: The choice of the surgical technique consisted first in the creation of a buccal fat pad to restore the important lack of tissue over the underlying bones, then in the positioning of a dermal acellular matrix. Three weeks later, once the neodermal formation was finished, a split thickness graft was placed. Results: This is a not yet described association that represents a good surgical option for the restoration of large cheeck defects that allows good functional and cosmetic result in older patient when minimal surgical invasion and operative duration are necessary because of a patient’s general condition. The post-operative course with this surgical technique was regular and a good functional result was achieved. Conclusions: This technique provides an adequate functional coverage, a restoration of soft tissue lacking and an acceptable cosmetic result without ectropion.

2017 - The GENDER ATTENTION Observational Study: Gender and Hormonal Status Differences in the Incidence of Adverse Events During Cyclosporine Treatment in Psoriatic Patients [Articolo su rivista]
Colombo, Delia; Banfi, Giuseppe; Cassano, Nicoletta; Graziottin, Alessandra; Vena, Gino Antonio; Fiori, Giovanni Gualberto; Zagni, Emanuela; Stingeni, Luca; Chimenti, Sergio; Berardesca, Enzo; Micali, Giuseppe; Albertini, Giuseppe; De Simone, Clara; Bellia, Gilberto; Amato, Salvatore; Ayala, Fabio; Bardazzi, Ferderico; Bernengo, Maria Grazia; Bongiorno, Maria Rita; Borroni, Giovanni; Calvieri, Stefano; Pinton, Piergiacomo Calzavara; Cannarozzo, Calcedonio; Cannavò, Serafinella Patrizia; Cattaneo, Angelo; Ceschini, Rossella; Colombo, Enrico; Congedo, Maurizio; Cusano, Francesco; De Pità, Ornella; Di Nuzzo, Sergio; Donadio, Dario; Donelli, Stefano; Filotico, Raffaele; Flori, Maria Laura; Germino, Maurizio; Girolomoni, Giampiero; Kokelj, Franco; Lanzoni, Anna; Lembo, Giuseppe; Locatelli, Andrea; Magnoni, Cristina; Marconi, Marco; Motolese, Alberico; Papini, Manuela; Parodi, Aurora; Pau, Monica; Peris, Ketty; Piaserico, Stefano; Piccirillo, Angelo; Potenza, Concetta; Ricotti, Giuseppe; Sedona, Patrizio; Stinco, Giuseppe; Valenti, Giancarlo; Fornasa, Cleto Veller; Zichichi, Leonardo; Ori, Alessandra; Amici, Christian; Zullo, Alessandro; Simoni, Lucia; Lanci, Nicole; Longagnani, Chiara; Sala, Saide; Trevisan, Francesca; Magri, Giovanna
abstract

Introduction: Female sex has been shown to be a risk factor for the development of adverse drug reactions; however, this has not been studied for cyclosporine (CsA). The aim of this study was to investigate, in Italian dermatological practice, the influence of gender and menopause and related hormones on the incidence of adverse events (AEs) during CsA treatment in psoriatic patients. Methods: Multicenter, prospective, observational study conducted from May 2011 to June 2013. Patients with plaque psoriasis, undergoing a new CsA administration course, or about to start it, were enrolled in the outpatient clinics of Italian dermatological centers. During the 2–6 months of study duration, patients had to note all AEs that occurred in a diary that was reviewed by the investigators at the follow-up visit. Sex hormone levels were measured within 7 days from the start date of a menstrual cycle. Results: A total of 969 adult psoriatic patients were enrolled in the study, divided into four cohorts: fertile women and corresponding age-matched men; postmenopausal women and corresponding age-matched men. A significant difference in the percentage of patients with AEs was observed between fertile and postmenopausal women, but not between women and age-matched men. AE incidence rate was about 37% higher in fertile women than in age-matched men and about 18% higher in postmenopausal women than in age-matched men, but differences were not statistically significant. Incidence rate ratio of fertile vs. postmenopausal women was 0.67, reaching statistical significance. AEs were mild or moderate in severity in the great majority of patients of all cohorts and postmenopausal women had significantly less grade 1–2 AEs compared to fertile women, but more grade 3–4 AEs. FSH levels were significantly higher in postmenopausal women reporting no AEs, and DHEA sulfate levels were about 10% higher in men with no AEs, compared to those reporting at least one AE. Cortisol levels were slightly though significantly higher in postmenopausal women with no AE. Conclusions: A better understanding of sex- and hormone-related influences on drug responses may help to improve drug safety and efficacy, by permitting one to tailor pharmacological treatments to individual subjects or defined patient cohorts. Funding: Novartis Farma S.p.A., Italy.

2016 - Crescentic flap for the reconstruction of the nose after skin cancer resection [Articolo su rivista]
Arginelli, Federica; Salgarelli, Attilio Carlo; Ferrari, Barbara; Losi, Amanda; Bellini, Pierantonio; Magnoni, Cristina
abstract

Purpose Non-melanoma skin cancers are the most frequent skin tumours; in 25.5% of cases, they are reported to affect the nasal area. For an excellent surgical outcome, first of all the radical excision of the lesion is important, with appropriate margins of healthy skin in order to avoid recurrences. Moreover is important to achieve a good aesthetical result, avoiding distortion of the aesthetic units and preserving their functions. Material and methods We have applied the modified crescentic flap, described by Smadja in 2007, to 24 nasal skin defects left by oncologic surgery. It consists of the crescent-shaped resection of Burow's triangle all around the alar groove that allows the advancement of the flap to the tip of the nose, hiding the scar in the alar groove. Results The outcome and the long-term follow-up were completely satisfactory both for patients and for surgeons. Conclusion For skin defects localized in the midline or paramedian line of the dorsum of the nose, the crescentic flap seems to be a good solution to obtain the better aesthetic result with respect to both anatomy and function of the nasal area, sparing the patient a second intervention or an overly invasive procedure.

2016 - Functionalized gold nanoparticles for topical delivery of methotrexate for the possible treatment of psoriasis [Articolo su rivista]
Bessar, Hagar; Venditti, Iole; Benassi, Luisa; Vaschieri, Cristina; Azzoni, Paola; Pellacani, Giovanni; Magnoni, Cristina; Botti, Elisabetta; Casagrande, Viviana; Federici, Massimo; Costanzo, Antonio; Fontana, Laura; Testa, Giovanna; Mostafa, Fawzia Farag; Ibrahim, Samia Ali; Russo, Maria Vittoria; Fratoddi, Ilaria
abstract

Gold nanoparticles (AuNPs) represent an effective choice for topical drug delivery systems thanks to their small size, general non-toxicity, ease of functionalization and high surface to volume ratio. Even if systemic, methotrexate still plays an important role in psoriasis treatment: its topical use shows insufficient percutaneus penetration owing to limited passive diffusion, high molecular weight and dissociation at physiological pH. The aim of our study was to design a new drug delivery nanocarrier for Methotrexate and to improve its solubility, stability and biodistribution. AuNPs were on purpose prepared with a hydrophilic stabilizing layer, in order to improve the colloidal stability in water. Water-soluble gold nanoparticles functionalized by sodium 3-mercapto-1-propansulfonate (Au-3MPS) were prepared and loaded with methotrexate (MTX). The loading efficiency of MTX on Au-3MPS was assessed in the range 70-80%, with a fast release (80% in one hour). The release was studied up to 24h reaching the value of 95%. The Au-3MPS@MTX conjugate was fully characterized by spectroscopic techniques (UV-vis, FTIR) and DLS. Preliminary toxicity tests in the presence of keratinocytes monolayers allowed to assess that the used Au-3MPS are not toxic. The conjugate was then topically used on C57BL/6 mouse normal skin in order to trace the absorption behavior. STEM images clearly revealed the distribution of gold nanoparticles inside the cells. In vitro studies showed that Methotrexate conjugated with Au-3MPS is much more efficient than Methotrexate alone. Moreover, DL50, based on MTT analysis, is 20 folds reduced at 48 h, by the presence of nanoparticles conjugation. UV-vis spectra for in vivo tracing of the conjugate on bare mouse skin after 24h of application, show increased delivery of Methotrexate in the epidermis and dermis using Au-3MPS@MTX conjugate, compared to MTX alone. Moreover we observed absence of the Au-3MPS in the dermis and in the epidermis, suggesting that these layers of the skin do not retain the nanoparticles. Based on our data, we found that the novel Au-3MPS@MTX conjugate is an effective non-toxic carrier for the satisfactory percutaneous absorption of Methotrexate and could help in possible topical treatment of psoriasis.

2016 - Guidance flap choice for lip cancer: Principles, timing and esthetic-functional results [Articolo su rivista]
Salgarelli, Attilio Carlo; Setti, Giacomo; Bellini, Pierantonio; Magnoni, Cristina; Robiony, Massimo; Consolo, Ugo
abstract

The purpose of this study is to help in the choice of an appropriate reconstructive technique by reference to the dimensions of the defect, the required functional and esthetic outcomes, and retention of adequate surgical safety margins to prevent primary tumor recurrence. Material and methods: A total of 158 patients were treated. We indicate how the most appropriate reconstructive method may be chosen, with reference to the size and position of the cancer and depth of tissue infiltration. Result: Of all patients, 89 (56.3%) had T1 (lesions up to 2 cm long, less than 1/3). The remaining patients had T2 lesions >2 cm, from 1/3 to 2/3 of lip involvement (50 patients), T3 lesions >4 cm, more than 2/3 of lip involvement (18), and a T4 lesion > 5.5 cm with commissure involvement (1). Conclusion: We share the widespread view that a surgeon who performs a reconstruction using the minimal tissue components required to close the lesion will achieve the best results. Reconstruction does not influence prognosis and overall should be oriented to the defect. Careful, clean, and safe resection of lip carcinoma, with creation of healthy margins, can be followed by functional and esthetic lip reconstruction.

2016 - NMR Contribution to the comprehension of metabolic disorders in inflammatory and neoplastic hyper-proliferative disease [Abstract in Rivista]
Righi, V.; Mucci, Adele; Fazio, Nicola; Magnoni, Cristina
abstract

A progression model based on genetic alterations has been proposed for several types of tumours and in skin, the concept of cancerization field involves cluster of genetically altered cells in a chronically photo damaged skin without clinical evidence of neoplastic lesions1. Cutaneous Squamous Cell Carcinoma (CSCC) is a common skin cancer characterized by malignant proliferation of keratinocytes. CSCC usually arises from precursor lesions such as actinic keratosis (AKs), but can also grow de novo or on chronically inflamed skin. AKs are the most common skin lesion of disordered keratinocyte proliferation in the disease continuum of photodamaged skin that may lead to invasive CSCC. Although there is reason to believe AKs convert to CSCC, there is also some evidence against this. Most AKs do not progress to CSCC. Clinically, some CSCC appear to arise de novo in healthy skin and in support of this, different genetic abnormalities may underlie AKs and CSCC. Exploring the metabolome of cancer and precancerous lesions seems to be a parallel and effective way to understand the phenotypic changes associated with cancer progression. Metabolomics could reveal novel cancer biomarkers that might expand our current understanding of the multi-factorial disease. Nuclear Magnetic Resonance (NMR) spectroscopy, giving an accurate description of the molecular composition of a human tissue provides a “fingerprint” of the whole metabolome.2 Correlations between some metabolites and proliferative markers allow gaining insight into the relationship between cellular proliferation and metabolic changes associated with the presence of tumor and its aggressiveness.

2016 - Skin cancer risk in outdoor workers: A European multicenter case-control study [Articolo su rivista]
Trakatelli, M.; Barkitzi, K.; Apap, C.; Majewski, S.; De Vries, E.; Coebergh, Jan Willem; Apalla, Zoe; Ioannides, Dimitrios; Kalabalikis, Dimitrios; Kalokasidis, Kostantinos; Kitsou, Anastasia; Siskou, Stella; Traianou, Athina; Sotiriadis, Dimitrios; Moreno-Ramirez, David; Ferrandiz, Lara; Ruiz-De-Casas, Andres; Micallef, Rita; Ranki, Annamari; Saksela, Olli; Pitkänen, Sari; Stockfleth, Eggert; Altsitsiadis, Efthimios; Hinrichs, Birgit; Ulrich, Martina; Magnoni, Cristina; Fiorentini, Chiara; Proby, Charlotte; Crawford, Leaca
abstract

Background Exposure to ultraviolet radiation (UVR) is the most important external risk factor for skin cancer. Outdoor workers, who are exposed to high ambient UVR levels are at increased risk. Objective To compare outdoor with indoor workers in terms of: (i) skin cancer risk factors, and (ii) risk of developing skin cancer. Methods Using descriptive methods and a large multicenter European case-control study, we compared risk factor patterns between outdoor (N = 1416) and indoor workers (N = 1863). Risk of developing basal cell carcinoma (BCC), squamous cell carcinoma (SCC), melanoma and actinic keratosis (AK) were analysed by type of work using multivariate logistic regression models, for three categories of work: indoor; farming/construction; other outdoor work. Results Although skin phototype was equally distributed by type of work, significantly less outdoor than indoor workers used sunscreen in their own country (44.3% vs. 60.2%), but had more outdoor hobbies (66.2% vs. 58.2%). Outdoor workers had lower educational levels, and felt less confident in understanding medical information and filling medical forms (all P < 0.001). Outdoor workers had more signs of photodamage (78.1% vs. 65.5%) and among the skin cancer patients, 37.7% of outdoor workers vs. 28.6% of indoor workers had ≥2 skin cancers diagnosed during their lifetime. Multivariate logistic regression models showed significantly increased risk of outdoor vs. indoor work for AK (ORother outdoor= 1.55, ORfarming/construction= 2.58), SCC (ORother outdoor= 1.32, ORfarming/construction= 2.77) and BCC (ORother outdoor= 1.53, ORfarming/construction= 1.83). No significant associations were found for melanoma. The risk of all types of skin cancer and AK was significantly increased for workers with ≥5 years of outdoor work. Conclusions Outdoor workers had more risk behaviour with similar constitutional skin cancer risk factors: more UV exposure (both occupational and leisure) and less sunscreen use and lower health literacy. This results in higher exposure, more photodamage and an increased risk of developing AK, BCC and SCC.

2015 - Sentinel lymph node biopsy in lower lip cancer [Abstract in Rivista]
Setti, G.; Salgarelli, Attilio Carlo; Magnoni, Cristina; Bellini, Pierantonio; Consolo, Ugo
abstract

Objective Lower lip SCC neck nodes metastases increase mortality to 50% of cases when involved. Recent NCCN guidelines don't refer to occult metastases investigation strategy. We report application and results of sentinel node biopsy as diagnostic method in cN0 lesions. Study Design Through the use of lymphoscintigraphy, performed with colloidal 99Tc injected in the main lesion, 22 patients with neck cN0 and negative ultrasonography were treated. Tumor staging varies from T1 (77,3%) to T4. Results Only two cases, both T2, result positive to micro metastases investigation, realized with monoclonal anti-body MNF 116 marking and 200μm microtome sections; either undergo to ND which allowed to identify respectively 1 pN+ each, same neck level of sentinel node. Conclusions To avoid needless elective neck dissection or a delay of treatment in occult N+ cases, biopsy method can identify metastases presence in cN0 cases, though we suggest expanding the method also to all T1 less than 1 cm.

2015 - Una strana placca eritematosa parzialmente ulcerata [Relazione in Atti di Convegno]
Mandel, Victor Desmond; Fiorentini, Chiara; Benati, Elisa; Magnoni, Cristina; Pellacani, Giovanni
abstract

Presentiamo il caso di una donna di 38 anni affetta da sclerosi tuberosa ed in terapia immunosoppressiva dal 2006 per trapianto renale. La paziente presentava grave deficit cognitivo e di coordinazione motoria. All’esame obiettivo si rilevava la presenza di una placca eritematosa in regione pretibiale sinistra, parzialmente ulcerata, di cui la paziente riferiva comparsa e persistenza da circa 3 anni. Nel resto del corpo la cute si presentava integra eccetto alcune lesioni da grattamento. In passato erano state eseguite due biopsie cutanee per un miglior inquadramento diagnostico il cui referto istologico era risultato completamente aspecifico. La madre riferiva di aver notato la comparsa talvolta di alcune bolle sul corpo, che si rompevano e guarivano spontaneamente senza lasciare esiti cicatriziali. Nel sospetto di una patologia bollosa si eseguiva un prelievo ematico per immunofluorescenza indiretta e due biopsie cutanee per esame istologico e immunofluorescenza diretta. Al momento della consegna degli esiti degli esami, che deponevano per un quadro di pemfigoide bolloso, si osservava un’intensa area disepitelizzata nella regione pretibiale sinistra e due piccole bolle tese sulla gamba destra. Localmente si applicavano steroide topico sulle lesioni bollose e medicazione a base di idrofibra con argento sull’area pretibiale sinistra con risoluzione del quadro cutaneo.

2014 - Dermoscopic yellow structures in basal cell carcinoma [Articolo su rivista]
Bellucci, Claudia; Arginelli, Federica; Bassoli, Sara; Magnoni, Cristina; Seidenari, Stefania
abstract

BACKGROUND: Yellowish structures in dermoscopy comprise milia-like cysts (MLCs) and yellow lobular-like structures. OBJECTIVE: This study aimed at establishing the frequency of these features in BCC and at describing their dermoscopic details. METHODS: A retrospective analysis of digital dermoscopic images referring to 400 BCCs was performed. Images were evaluated for the presence of starry and cloudy MLCs and yellow lobular-like structures. RESULTS: Among the 400 BCCs constituting our database, 40 presented yellowish structures (10%). "Yellow" BCCs were located more frequently on the head and were mainly of the nodular type. MLCs were observed in 7.75% of the cases (with a mean number of 4.9 MLCs per lesion), whereas yellow globules were noticed in 4.2% /ucodep of the lesions. CONCLUSION: In the presence of BCC specific dermoscopic criteria, the observation of MLCs and yellow lobular-like structures should not prompt the dermatologist to exclude the diagnosis of BCC

2014 - Dermoscopy of small melanomas: just a miniaturized dermoscopy? [Articolo su rivista]
Ferrari, C; Seidenari, Stefania; Borsari, S; Fabiano, A; Bassoli, Sara; Giusti, F; Ponti, Giovanni; Magnoni, Cristina
abstract

BACKGROUND AND OBJECTIVE: Small melanomas (MMs) are usually MMs in an initial growth phase, deserving attention by the clinician aiming at an early diagnosis. The aim of this study was to identify clues for early diagnosis of small MM, by comparing the dermoscopic features of MMs smaller than 4 mm (micromelanomas) to those of larger MMs. MATERIALS AND METHODS: The database consisted in the dermoscopic images of 482 MMs which were retrieved and measured digitally. The ABCD and the 7-point criteria were evaluated on the whole data base by 3 expert dermoscopists, whereas the main dermoscopic pattern was assessed only on micromelanomas. The dermoscopic aspects were correlated to clinical and histological ones. RESULTS: Most 7-point and ABCD scores and criteria referring to micromelanomas differed from those of the whole MM data base. Lesion asymmetry, number of colors, blue-whitish veil, atypical vessels, irregular globules/dots and regression increased according to MM diameter. An inverse trend was observed for atypical network and irregular pigmentation, which were more frequently observed in micromelanomas than in larger ones. Among the 22 micromelanomas, 12 lesions were in situ, whereas the other 10 were 0.2-2 mm thick. The clinical and dermoscopic characteristics of the 2 groups were similar. CONCLUSIONS: Micromelanomas are not a rarity. However, the clinician should be aware of the fact that the majority of them lack most of the dermoscopic features presented by larger lesions. This article is protected by copyright. All rights reserved.

2014 - Functional classification of wound dressings [Articolo su rivista]
Greco, A.; Mastronicola, D.; Magnoni, C.
abstract

2014 - High Magnification Digital Dermoscopy of Basal Cell Carcinoma: A Single-centre Study on 400 cases. [Articolo su rivista]
Seidenari, S; Bellucci, C; Bassoli, S; Arginelli, F; Magnoni, Cristina; Ponti, Giovanni
abstract

The aim of this study was to assess the frequency of classic dermoscopic basal cell carcinoma (BCC) features and the sensitivity of new descriptors, such as light brown nests (homogeneous and structured) only visible employing a high magnification digital videomicroscope. A retrospective analysis of 2,024 highly magnified digital images referring to 400 BCCs was performed by 3 independent observers, who assessed 11 classic BCC descriptors and the new ones. Light brown nests were detected in 40.5% of BCCs. Homogeneous ones were observable in 17.8%, and structured nests in 32.8%. Light brown nests were visible in 14.3% of non-pigmented lesions, whereas in the pigmented groups these were observed in 42-54% of the cases. We suggest that brown nests described in this study may improve early recognition of superficial BCCs and of non-pigmented or slightly pigmented ones that may lack classic dermoscopic patterns.

2014 - Sentinel lymph node biopsy in lower lip cancer [Abstract in Rivista]
Setti, C.; Salgarelli, Attilio Carlo; Magnoni, Cristina; Bellini, Pierantonio; Consolo, Ugo
abstract

Vedasi l'allegato

2014 - Stem cell properties in cell cultures from different stage of melanoma progression. [Articolo su rivista]
Magnoni, Cristina; S., Guidice; Pellacani, Giovanni; G., Bertazzoni; Longo, Caterina; E., Veratti; D., Morini; L., Benassi; S., Al Jalbout; C., Vaschieri; P., Azzoni; DE POL, Anto; S., Seidenari; Tomasi, Aldo; Ponti, Giovanni
abstract

Cutaneous Melanoma is an extremely heterogeneous human cancer. The most aggressive melanoma may contain deregulated cells with undifferentiated/stem cell-like phenotype. A critical mechanism by which melanoma cells enhance their invasive capacity is the dissolution of the intercellular adhesions and the acquisition of mesenchymal features as a part of an epithelial-to-mesenchimal transition (EMT). The aim of this study was to clarify the role of stem cell-like population in human melanomas through the in vitro analysis of melanocytic cell cultures, obtained from distinct histotypes of primary and metastatic malignant melanoma. Patients with advanced melanoma larger than 2cm diameter and/or wider than 300mm2 surface were enrolled. The melanoma cells were isolated from skin biopsies of lentigo melanoma (LM), superficial spreading (SS), nodular melanoma (NM) and metastatic melanoma (MM) and maintained in different media in order to evaluate the colony-forming unit assay, alkaline phosphatase and toluidine blue stain and the cell ability to differentiate into osteogenic and adipogenic lineages. Immunohistochemistry and flow cytometry analysis were performed in order to evaluate antigenic markers CD90, CD73, CD105, CD146, CD20, CD166 and nestin. This study confirms that melanoma can include an heterogeneous cell population with both abilities to self-renew and to give rise to differentiated progeny. Melanoma cells displayed the intra-tumoral heterogeneity and dynamic antigen phenotypes. Histologically, the transitions from normal skin to DN to LM to NM to MM was associated with a gradual increase in the expression of CD146, CD20, CD133, Nestin and CD73 by melanoma cells. These molecular evidences could be a milestone for the development of novel biomolecular targeted-therapy approaches.

2014 - Un eritema anulare centrifugo ricorrente da molti anni in estate [Poster]
Mandel, Victor Desmond; Manfredini, Marco; Ferrari, Barbara; Giusti, Francesca; Magnoni, Cristina; Pellacani, Giovanni
abstract

L’eritema anulare centrifugo ricorrente annualmente è una peculiare variante clinica di eritema anulare centrifugo superficiale, clinicamente ed istologicamente indistinguibile da questo, ma caratterizzato da una ricorrenza annuale e stagionale. Presentiamo il caso di una donna di 46 anni con un’importante sintomatologia pruriginosa. All’esame obiettivo si rilevava la presenza di chiazze eritematose con risoluzione centrale multiple e confluenti agli arti superiori ed inferiori. Il cavo orale, faccia, tronco, mani e piedi erano risparmiati dalle lesioni. La paziente non assumeva farmaci, non era affetta da patologie sistemiche e riferiva che tali manifestazioni si presentavano ogni anno, da 8 anni a questa parte, nel periodo estivo e regredivano spontaneamente in autunno. La diagnosi differenziale dell’eritema anulare centrifugo comprende principalmente il granuloma anulare, micosi fungoide, pitiriasi rosea, tinea corporis, sarcoidosi, orticaria e lebbra. Clinicamente le manifestazioni cutanee deponevano per un quadro di eritema anulare centrifugo. Veniva eseguita una biopsia cutanea il cui quadro istologico, caratterizzato da un infiltrato infiammatorio perivascolare linfoistiocitario nel derma papillare e medio, confermava la diagnosi di eritema anulare centrifugo. Mediante l’esecuzione di una completa serie di esami di laboratorio e strumentali veniva esclusa la presenza di una patologia sottostante. Veniva impostata una terapia sistemica steroidea con risoluzione del quadro cutaneo.

2014 - Wave technique in lower lip cancer [Abstract in Rivista]
Bellini, Pierantonio; Salgarelli, Attilio Carlo; Setti, G.; Magnoni, Cristina; Consolo, Ugo
abstract

Vedasi l'allegato

2013 - Are the neck malignant melanomas different from the ones affecting the head? Clinicopathologic, dermoscopic and prognostic findings [Articolo su rivista]
Ponti, Giovanni; Pollio, Annamaria; Tomasi, Aldo; Borsari, Stefania; Magnoni, Cristina; Mandel, Victor Desmond; Loschi, Pietro; Ruini, Cristel; Seidenari, Stefania
abstract

Background: Malignant melanomas of the head and neck are usually considered as a unique entity in comparison to other body sites. However, no characterization of neck melanoma has been performed so far, despite the clear anatomic and histological differences. Aim: We investigated clinical, demographic, histological and dermoscopic differences between face, scalp and neck melanoma. Materials and methods: A retrospective analysis of medical and histologic records from 116 melanomas of the head and neck area collected between January 2003 and January 2008 was performed. Body site, gender, age, number of lesions, age at first melanoma diagnosis, size, Clark level, association with nevi, presence or absence of mitoses and/or ulceration, presence of synchronous and/or metachronous melanoma were recorded. Moreover, digital dermoscopy images of 92 melanomas of the head and neck area were analyzed for main dermoscopic patterns and lesion diameter. Results: Significant differences in Breslow thickness, ex-naevo origin and tumor size among neck and face-scalp melanomas were observed. Neck MM patients were younger than those with MM of face and scalp. In contrast to scalp and face, no patient died from neck melanoma. Dermoscopic patterns were similar to those of trunk-limbs MM, and no lesion showed a lentigo maligna pattern which was observed in most lesions of the face. Conclusion: Melanomas of the neck must be distinguished from face and scalp melanomas because of younger age, different dermoscopic patterns and ex-naevo origin and better prognosis. These data should be taken into account both from an epidemiological and clinical point of view.

2013 - Electrical stimulation as adjuvant treatment for chronic leg ulcers of different aetiology:an RCT [Articolo su rivista]
Magnoni, Cristina; Rossi, Elena; Fiorentini, Chiara; Baggio, Andrea; Ferrari, Barbara; Giannetti, Alberto
abstract

OBJECTIVE: To investigate the effectiveness of an innovative electrical stimulation (ES) therapy as adjuvant treatment for chronic wounds of various aetiology, in terms of pain and ulcer healing. METHOD: Patients with chronic limb ulcers were enrolled for the study and randomised into the intervention or control group. The intervention group received conventional treatment plus ES therapy (FREMS; Lorenz Lifetech) while the control group received only conventional treatment. Each ES treatment cycle consisted of 12 sessions performed in 4 weeks (three sessions/week). All patients were treated until full wound healing occurred, or for a maximum of 9 ES cycles, with a 2-week rest between each cycle. RESULTS: A total of 60 patients were enrolled in the study and randomised into the two groups: the intervention group (n=30) and the control group (n=30). During follow-up, some patients terminated the protocol because they reached the ulcer closure before the maximum of 9 cycles. The analysis of the effect of ES on pain and ulcer healing was performed on all patients who underwent at least two consecutive clinical evaluations (two cycles), in order to reach a compatible sample size with the primary objective (one patient withdrew). In both groups, there was a significant reduction of pain compared with baseline (p < 0.05), starting from T6 visit in the first cycle. In particular, there was a significant reduction of pain in the intervention group compared with the control group after 14 days, and this reduction continued until the end of the second cycle. Similarly, there was a significant reduction of PUSH tool score in the intervention group compared with the control group after 14 days, and this reduction continued until the end of the second cycle. CONCLUSION: Data collected in this study support data in the literature. Analysis of longitudinal data analysed by simple models and complex models suggest that the ES therapy had a positive and significant effect on pain reduction (VAS) and on the improvement of ulcer healing process in terms of the PUSH tool total index compared with conventional treatment, and may have induced a significant acceleration of the wound-healing process.

2013 - High-resolution multiphoton tomography and fluorescence lifetime imaging of UVB-induced cellular damage on cultured fibroblasts producing fibres [Articolo su rivista]
Stefania, S.; Simona, S.; Paola, A.; Luisa, B.; Stefania, B.; Jennifer, C.; Chiara, F.; Paul, F.; Stefania, G.; Karsten, K.; Cristina, M.; Clifford, T.; Christopher, D.
abstract

Background: Multiphoton tomography (MPT) is suitable to perform both ex vivo and in vivo investigations of living skin and cell cultures with submicron resolution. Fluorescence lifetime imaging (FLIM) generates image contrast between different states of tissue characterized by various fluorescence decay rates. Our purpose was to combine MPT and FLIM to evaluate fibroblasts and collagen fibres produced in vitro. Methods: Fibroblast cultures, 2-4 days old, at a subconfluent stage, were evaluated before and after irradiation with a single UVB dose. One month old cultures stimulated with ascorbic acid were also assessed. Results: After UVB radiation, fibroblasts appear irregular in size, lose their alignment and show a decrease in fluorescence lifetime. One month-old fibroblasts, producing collagen fibres after stimulation with ascorbic acid, appear as small roundish structures intermingled by filaments showing a granular arrangement. Conclusion: The combination of MPT and FLIM may be useful for the in vitro study of cell modifications induced by injurious or protective agents and drugs. © 2013 John Wiley &amp; Sons A/S.

2013 - Multiphoton laser tomography and fluorescence lifetime imaging of melanoma: morphologic features and quantitative data for sensitive and specific non-invasive diagnostics. [Articolo su rivista]
Seidenari, S; Arginelli, F; Dunsby, C; French, Pm; König, K; Magnoni, Cristina; Talbot, C; Ponti, Giovanni
abstract

Multiphoton laser tomography (MPT) combined with fluorescence lifetime imaging (FLIM) is a non-invasive imaging technique, based on the study of fluorescence decay times of naturally occurring fluorescent molecules, enabling a non-invasive investigation of the skin with subcellular resolution. The aim of this retrospective observational ex vivo study, was to characterize melanoma both from a morphologic and a quantitative point of view, attaining an improvement in the diagnostic accuracy with respect to dermoscopy. In the training phase, thirty parameters, comprising both cytological descriptors and architectural aspects, were identified. The training set included 6 melanomas with a mean Breslow thickness±S.D. of 0.89±0.48 mm. In the test phase, these parameters were blindly evaluated on a test data set consisting of 25 melanomas, 50 nevi and 50 basal cell carcinomas. Melanomas in the test phase comprised 8 in situ lesions and had a mean thickness±S.D. of 0.77±1.2 mm. Moreover, quantitative FLIM data were calculated for special areas of interest. Melanoma was characterized by the presence of atypical short lifetime cells and architectural disorder, in contrast to nevi presenting typical cells and a regular histoarchitecture. Sensitivity and specificity values for melanoma diagnosis were 100% and 98%, respectively, whereas dermoscopy achieved the same sensitivity, but a lower specificity (82%). Mean fluorescence lifetime values of melanocytic cells did not vary between melanomas and nevi, but significantly differed from those referring to basal cell carcinoma enabling a differential diagnosis based on quantitative data. Data from prospective preoperative trials are needed to confirm if MPT/FLIM could increase diagnostic specificity and thus reduce unnecessary surgical excisions.

2013 - Negative Pigment Network Identifies a Peculiar Melanoma Subtype and Represents a Clue to Melanoma Diagnosis: A Dermoscopic Study of 401 Melanomas [Articolo su rivista]
Bassoli, Sara; Ferrari, Chiara; Borsari, Stefania; Giusti, Francesca; Magnoni, Cristina; Pellacani, Giovanni; Seidenari, Stefania
abstract

The dermoscopic descriptor "negative pigment network" (NPN) has been reported in several types of melanocytic and non-melanocytic lesions, although it has a higher frequency of association with melanoma and Spitz naevus. In a study of 401 consecutive melanomas, excluding facial, acral and mucosal locations, the frequency and variability of NPN were investigated, and the results of NPN correlated with clinical and histopathological data. NPN of any extension was found in 27% of melanomas, most frequently invasive and arising from a naevus on the trunk of young subjects. Seven percent of melanomas in the study population showed presence of NPN in more than half of the lesion area; most of these did not show typical dermoscopic melanoma features. The authors propose a new melanoma subtype, in which extensive NPN should be considered as a diagnostic indicator.

2013 - Proteomic Analysis of PTCH1+/- Fibroblast Lysate and Conditioned Culture Media Isolated from the Skin of Healthy Subjects and Nevoid Basal Cell Carcinoma Syndrome Patients. [Articolo su rivista]
Ponti, Giovanni; Bertazzoni, G; Pastorino, L; Monari, Emanuela; Cuoghi, Aurora; Bergamini, Stefania; Bellei, Elisa; Benassi, Luisa; Azzoni, Paola; Petrachi, Tiziana; Magnoni, Cristina; Pellacani, Giovanni; Loschi, P; Pollio, A; Witkowski, Am; Tomasi, Aldo
abstract

Background. The pathogenesis underlying the increased predisposition to the development of basal cell carcinomas (BCCs) in the context of Gorlin-Goltz syndrome is linked to molecular mechanisms that differ from sporadic BCCs. Patients with Gorlin syndrome tend to develop multiple BCCs at an early age and present with tumors of non-sun-exposed skin. The aim of this study was to compare the proteomic profile of cultured fibroblast and fibroblast conditioned culture media of PTCH1+ and nonmutated fibroblasts. Results. Proteomic analysis was performed using Surface-Enhanced Laser Desorption/Ionization Time-of-Flight mass spectrometry in PTCH1+ fibroblast conditioned media isolated from not affected sun-protected skin areas of Gorlin patients and from healthy subjects. 12 protein cluster peaks, >5 kDa, had significant differences in their peak intensities between PTCH1+ and PTCH1- subject groups. We detected a strongly MMP1 overexpression in PTCH1+ fibroblasts obtained from NBCCS patients with respect to healthy donors. Conclusion. Protein profiles in the fibroblast conditioned media revealed statistically significant differences between two different types (missense versus nonsense) of PTCH1 mutations. These differences could be useful as signatures to identify PTCH1 gene carriers at high risk for the development of NBCCS-associated malignancies and to develop novel experimental molecular tailored therapies based on these druggable targets.

2013 - Relationship between histological and computer based assessment of melanoma diameter and thickness in head & neck vs. trunk melanoma. [Articolo su rivista]
Seidenari, Stefania; A., Fabiano; S., Al Jalbout; S., Bassoli; S., Borsari; Magnoni, Cristina; Tomasi, Aldo; Vinceti, Marco; Ponti, Giovanni
abstract

BackgroundDiameter represents a controversial parameter in the diagnosis of melanoma (MM). The ABCD rule considers it an important diagnostic parameter, while some authors decrease its relevance. Also, its measurement is not always reliable, histological methods being usually used. The current study aims to compare histological and digital measurements and to evaluate the correlation between MM head- and trunk diameter and thickness. Methods The study population was subdivided according to diameter subgroups considering head- and trunk-limbs lesions separately. Digital diameters measured by an automatic software referring to 477 MM images were compared to diameters reported on the pathologist’s records. Clinical and histological information was also considered, and the correlation between diameter and thickness was assessed. Odds ratios (OR) were computed for different diameter subgroups. Results Mean digital diameters of head MMs and trunk limbs MMs exceeded histological measurements by 11% and 20%, respectively. In head MMs, no correlation between diameter and thickness was observable, whereas in the trunk-limbs group a direct relationship between thickness and diameter was noticed. ORs for non in situ vs in situ andfor ≥ 1 mm vs < 1 mm thick were low for small lesions, increasing for larger ones, indicating that the latter are more likely to be thick. ConclusionMM diameter should be assessed digitally to avoid tissue shrinkage after biopsy and imprecise in vivo measurements. Although nearly 10% of MMs might escape an early diagnosis based only on the D of the ABCD rule, MM diameter may be related to its thickness.The evaluation with a computer vision system should be recommended for small pigmented lesions (<6mm) in order to reduce the percentage of misdiagnosed smaller MMs and to better evaluate the parameter E of the ABCDE rule (Evolving lesion).

2012 - Assessing physicians' preferences on skin cancer treatment in Europe [Articolo su rivista]
Ferrandiz, L; Ruiz de Casas, A; Trakatelli, M; de Vries E, Ulrich; Aquilina, S; Saksela, O; Majewski, S; Ranki, A; Proby, C; Magnoni, Cristina; Pitkänen, S; Kalokasidis, K; Siskou, S; Hinrichs, B; Altsitsiadis, E; Stockfleth, E; Moreno Ramirez, D; Epiderm, Group
abstract

BACKGROUND: A wide variety of both surgical and nonsurgical therapies is currently available for patients with skin cancer. OBJECTIVES: This part of the EPIDERM (European Prevention Initiative for Dermatological Malignancies) project is aimed at the evaluation of the treatment preferences for skin cancer in eight countries of the European Union. METHODS: A multicentre hospital-based case-control study was carried out at dermatology departments in Finland, Germany, Greece, Italy, Malta, Poland, Scotland and Spain. Patients with skin cancer (basal cell carcinoma, actinic keratosis, squamous cell carcinoma, cutaneous malignant melanoma and Bowen disease) were consecutively enrolled between July 2008 and July 2010. Information on the study variables (sex, age, country, tumour type, anatomical location and treatment) was obtained from questionnaires designed by the EPIDERM project. RESULTS: In total, 1708 patients with skin cancer were included. Surgery was the first treatment option in 76·5% of the patients (P = 0·001). Actinic keratosis was the only tumour type in which nonsurgical treatment was more frequent than surgery (91·4%). Tumours on the head were less likely to be surgically excised than those at other locations (odds ratio 0·25, P = 0·001). Simple excision or curettage was the most common surgical procedure (65·4%), followed by graft and flaps (22·4%). Cryotherapy was the most common nonsurgical option (52·4%), followed by imiquimod (18·0%), photodynamic therapy (PDT; 12·0%), 5-fluorouracil (5-FU; 5·7%), and diclofenac with hyaluronic acid (4·0%). CONCLUSIONS: Surgery remains the first-choice treatment of skin cancer. Regarding nonsurgical treatments, the conservative treatments available (imiquimod, 5-FU, PDT and diclofenac gel) have not yet exceeded the use of ablative options such as cryotherapy despite their accepted benefit of treating field cancerization.

2012 - Basal cell carcinomas without histological confirmation and their treatment: an audit in four European regions. [Articolo su rivista]
Flohil, Sc; Proby, Cm; Forrest, A; van Tiel, S; Saksela, O; Pitkänen, S; Ahti, T; Micallef, R; de Vries, E; EPIDERM Group (Coebergh, Jw; Ioannides D, Kalabaliki D; Magnoni, Cristina; Fiorentini, Chiara; Majewski S, Crawford L.
abstract

BACKGROUND: Limited data are available on how often basal cell carcinomas (BCCs) are clinically diagnosed without histological confirmation and how they are treated. OBJECTIVES: Within the framework of the EPIDERM project, an audit was conducted in four European countries to study the occurrence of clinically diagnosed BCCs without histological confirmation and to investigate how these are treated. METHODS: In the Netherlands, Scotland, Finland and Malta studies were performed within different timeframes. Patients with one or more BCC(s) were selected and the number of clinically diagnosed BCCs without histological confirmation and their treatment was investigated by (manually) reviewing the (electronic) patient records and checking the (hospital) pathology databases to find evidence of histological confirmation. RESULTS: In the Netherlands, 1089 patients with a first histologically confirmed BCC developed 1974 BCCs of which 1833 (92·9%) were histologically confirmed and 141 (7·1%) were not. A 4-month retrospective study conducted in Scotland selected 294 patients with 344 BCCs; 306 (89·0%) were histologically confirmed and 38 (11·0%) were not. A 3-month prospective study performed at the same centre in Scotland identified 44 patients who developed 58 BCCs; 44 (75·9%) of these were histologically confirmed and 14 (24·1%) were not. In Finland, there were 701 patients who developed 977 BCCs, of which 807 (82·6%) were histologically and 170 (17·4%) nonhistologically confirmed. In Malta, there were 420 patients with 477 BCCs. Only three (0·7%) of them were clinically diagnosed without histological confirmation. In the Netherlands and Finland, clinically diagnosed BCCs without histological confirmation were most often treated with cryotherapy, whereas in Scotland 5% imiquimod cream was the preferred treatment modality. CONCLUSIONS: Although the frequency of clinically diagnosed BCCs without histological confirmation differed between the four European regions (range 0·7-24·1%), this confirms that the burden of BCC in Europe is underestimated when based on data from pathology and/or cancer registries.

2012 - Biological evaluation of MR36, a novel non-polyglutamatable thymidylate synthase inhibitor that blocks cell cycle progression in melanoma cell lines. [Articolo su rivista]
Giudice, Stefania; Benassi, Luisa; Bertazzoni, G; Veratti, E; Morini, D; Azzoni, Paola; Costi, Maria Paola; Venturelli, Alberto; Pirondi, Silvia; Seidenari, Stefania; Magnoni, Cristina
abstract

Melanoma is one of the most common cancers, and its incidence has continued to increase over the past few decades. Chemotherapy resistance and related defects in apoptotic signaling are critical for the high mortality of melanoma. Effective drugs are lacking because apoptosis regulation in this tumor type is not well understood. The folate pathway has been considered an interesting target for anticancer therapies, and approaches targeting this pathway have recently been extended to melanoma treatment. In this study, the intracellular apoptosis signaling pathways of two melanoma cells lines (SK-MEL-2 and SK-MEL-28) were investigated after treatment with a new experimental antifolate substance (MR36) that targets thymidylate synthase. In both melanoma cell lines, apoptosis induction was triggered by a p53-independent mechanism. MR36-induced apoptosis was associated with a loss of both mitochondrial membrane potential and caspase-3 activation. Induction of cell cycle arrest by MR36 was associated with changes in the expression of key cell cycle regulators, such as p21 and cyclin D1, and the hypophosphorylation of pRb. In addition, Fas signaling was also analyzed. These findings suggest that, unlike classical antifolates, MR36 exerted an inhibitory effect on both the enzymatic function and expression of thymidylate synthase, thereby inducing apoptosis through the activation of the extrinsic and intrinsic pathways in the melanoma cell lines. MR36 showed a different mechanism of action from the known antifolates (Nolatrexed and Pemetrexed) that resulted in higher anticancer activity. Therefore, MR36 should be included as a potential new therapeutic treatment in melanoma research.

2012 - Diagnosis of BCC by multiphoton laser tomography. [Articolo su rivista]
Seidenari, S.; Arginelli, F.; Bassoli, S.; Cautela, J.; Cesinaro, A. M.; Guanti, M.; Guardoli, D.; Magnoni, Cristina; Manfredini, M.; Ponti, Giovanni; König, K.
abstract

BACKGROUND/PURPOSE: Multiphoton Laser Tomography (MPT) is a non-linear optical technique that gives access to morphology and structure of both cells and extracellular matrix of the skin through the combination of autofluorescence imaging and second harmonic generation (SHG). The aim of this study was to identify MPT descriptors on ex vivo specimens of basal cell carcinoma (BCC) to assess the sensitivity and specificity of these criteria for the diagnosis of BCC and its differentiation from other skin tumours, inflammatory diseases and healthy skin.METHODS: In the preliminary study, MPT images referring to 24 BCCs and 24 healthy skin samples were simultaneously evaluated by three observers for the identification of features characteristic of BCC. In the main study, the presence/absence of the descriptors identified in the preliminary study was blindly evaluated on a test set, comprising 66 BCCs, 66 healthy skin samples and 66 skin lesions, including 23 nevi, 8 melanomas, 17 skin tumours and other skin lesions by 3 independent observers.RESULTS: In the preliminary study, three epidermal descriptors and six descriptors for BCC were identified. The latter included aligned elongated cells, double alignment of cells, cell nests with palisading and phantom islands. From the test set, 56 BCCs were correctly diagnosed, whereas in 10 cases the diagnosis was 'other lesions'. However, it was always possible to exclude the diagnosis of BCC in healthy skin and other lesion samples. Thus, overall sensitivity of the method was 84.85, whereas a specificity of 100% was observed with respect to both healthy skin and 'other lesions'.CONCLUSIONS: This study describes new morphological descriptors of BCC enabling its characterization and its distinction from healthy skin and other skin lesions in ex vivo samples, and demonstrates for the first time that MPT represents a sensitive and specific technique for the diagnosis of BCC.

2012 - Health literacy, sunscreen and sunbed use: an uneasy association [Articolo su rivista]
Altsitsiadis, E; Undheim, T; de Vries, E; Hinrichs, B; Stockfleth, E; Trakatelli, M; EPIDERM Group (Coebergh JW, Ioannides D; Magnoni, Cristina; Fiorentini, Chiara; Majewski S, Proby C.
abstract

BACKGROUND: Skin cancer can largely be prevented by avoiding unsafe ultraviolet radiation (UVR) exposure. The evidence on potential drivers of sunscreen and sunbed use is extensive, yet in some cases, such as education, remains rather unclear. Health literacy is receiving increasing attention, but its effect on tanning decisions has not yet been explored. OBJECTIVES: To explore the association between health literacy and tanning behaviour, in terms of sunscreen and sunbed use. METHODS: Self-reported data were collected through a common questionnaire in eight European countries under a common protocol. A three-item measure was used to assess health literacy; one item was collected to measure current sunscreen use and one item to measure current sunbed use. Descriptive statistics and analysis of variance tests were applied to explore the profile of sunbed and sunscreen users and health literacy among a number of variables. Univariate and multivariate logistic regressions were used to assess the relation between health literacy and sunscreen and sunbed use. RESULTS: Univariate results suggested that health literacy has opposite effects on sunscreen use vs. sunbed use. Increased health literacy was associated with the skin cancer protective practice of using sunscreen, but also with more sunbed use. In the multivariate models, health literacy had a significant effect only on sunscreen use. CONCLUSIONS: The findings suggest that health literacy can be an interesting approach for influencing sunscreen use. In the case of sunbeds, based on the findings and contrary to what was expected, it can be argued that interventions targeting health literacy seem less likely to reduce sunbed use. More research is needed to elucidate the effect of health literacy on sunscreen and sunbed use in order to improve UVR prevention strategies.

2012 - High resolution diagnosis of common nevi by multiphoton laser tomography and fluorescence lifetime imaging. [Articolo su rivista]
Arginelli, F; Manfredini, M; Bassoli, S; Dunsby, C; French, P; König, K; Magnoni, Cristina; Ponti, Giovanni; Talbot, C; Seidenari, S.
abstract

BACKGROUND: Multiphoton Laser Tomography (MPT) has developed as a non-invasive tool that allows real-time observation of the skin with subcellular resolution. MPT is readily combined with time resolved detectors to achieve fluorescence lifetime imaging (FLIM). The aim of our study was to identify morphologic MPT/FLIM descriptors of melanocytic nevi, referring to cellular and architectural features. METHODS: In the preliminary study, MPT/FLIM images referring to 16 ex vivo nevi were simultaneously evaluated by 3 observers for the identification of morphologic descriptors characteristic of melanocytic nevi. Proposed descriptors were discussed and the parameters referring to epidermal keratinocytes, epidermal melanocytes, dermo-epidermal junction, papillary dermis and overall architecture were selected. In the main study, the presence/absence of the specified criteria were blindly evaluated on a test set, comprising 102 ex vivo samples (51 melanocytic nevi, 51 miscellaneous skin lesions) by 2 observers. RESULTS: Twelve descriptors were identified: "short-lifetime cells in the stratum corneum", "melanin-containing keratinocytes", "dendritic cells", "small short-lifetime cells" in the upper and lower layers", "edged papillae", "non-edged papillae", "junctional nests of short-lifetime cells", "dermal cell clusters", "short-lifetime cells in the papilla", "monomorphic and regular histoarchitecture", "architectural disarray". CONCLUSION: Identified descriptors for benign melanocytic lesions proved sensitive and specific, enabling the differentiation between melanocytic nevi and non-melanocytic lesions.

2012 - Known and potential new risk factors for skin cancer in European populations: a multicentre case-control study [Articolo su rivista]
de Vries, E; Trakatelli, M; Kalabalikis, D; Ferrandiz, L; Ruiz de Casas, A; Moreno Ramirez, D; Sotiriadis, D; Ioannides, D; Aquilina, S; Apap, C; Micallef, R; Scerri, L; Ulrich, M; Pitkänen, S; Saksela, O; Altsitsiadis, E; Hinrichs, B; Magnoni, Cristina; Fiorentini, Chiara; Majewski, S; Ranki, A; Stockfleth, E; Proby, C; Epiderm, Group
abstract

BACKGROUND: During recent years numerous studies have suggested that personal and environmental factors might influence cancer development. OBJECTIVES: To investigate environmental and personal characteristics associated with skin cancer risk. METHODS: A multicentre hospital-based case-control study was performed in Finland, Germany, Greece, Italy, Malta, Poland, Scotland and Spain, including 409 patients with squamous cell carcinoma (SCC), 602 with basal cell carcinoma (BCC) and 360 with cutaneous malignant melanoma (CMM) and 1550 control persons. Exposures were assessed by questionnaires that were partly self-administered, partly completed by dermatologists. Unconditional logistic regression modelling was used to assess associations including the influence of certain drugs and food items on skin cancer risk. RESULTS: The usual associations were observed for sun exposure and pigmentation characteristics, with chronic sun exposure being most strongly associated with SCC risk, and naevi and atypical naevi with CMM risk. Use of ciprofloxacin was associated with a decreased risk of BCC [odds ratio (OR) 0·33] and use of thiazide diuretics was associated with an increased risk of SCC (OR 1·66). Ciprofloxacin was also associated with SCC (OR 0·34) and thiazines with BCC (OR 2·04), but these associations lost significance after correction for multiple testing. Consumption of pomegranate, rich in antioxidants, was associated with decreased BCC and SCC risk, also after correcting for multiple testing. Recent experience of stressful events was associated with increased risk, particularly of CMM. CONCLUSIONS: In this large case-control study from across Europe the expected associations were observed for known risk factors. Some new potential protective factors and potential risk factors were identified for consumption of certain food items, medication use and stress, which deserve further investigation in future studies.

2012 - Multiphoton laser tomography and fluorescence lifetime imaging of basal cell carcinoma: morphologic features for non-invasive diagnostics [Articolo su rivista]
Seidenari, Stefania; Arginelli, Federica; Dunsby, C; French, P; König, K; Magnoni, Cristina; Manfredini, Marco; Talbot, C; Ponti, Giovanni
abstract

Multiphoton laser tomography (MPT) combined with fluorescence lifetime imaging (FLIM) is a non-invasive imaging technique, which gives access to the cellular and extracellular morphology of the skin. The aim of our study was to assess the sensitivity and specificity of MPT/FLIM descriptors for basal cell carcinoma (BCC), to improve BCC diagnosis and the identification of tumor margins. In the preliminary study, FLIM images referring to 35 BCCs and 35 healthy skin samples were evaluated for the identification of morphologic descriptors characteristic of BCC. In the main study, the selected parameters were blindly evaluated on a test set comprising 63 BCCs, 63 healthy skin samples and 66 skin lesions. Moreover, FLIM values inside a region of interest were calculated on 98 healthy skin and 98 BCC samples. In the preliminary study, three epidermal descriptors and 7 BCC descriptors were identified. The specificity of the diagnostic criteria versus 'other lesions' was extremely high, indicating that the presence of at least one BCC descriptor makes the diagnosis of 'other lesion' extremely unlikely. FLIM values referring to BCC cells significantly differed from those of healthy skin. In this study, we identified morphological and numerical descriptors enabling the differentiation of BCC from other skin disorders and its distinction from healthy skin in ex vivo samples. In future, MPT/FLIM may be applied to skin lesions to provide direct clinical guidance before biopsy and histological examination and for the identification of tumor margins allowing a complete surgical removal.

2012 - Patched homolog 1 gene mutation (p.G1093R) induces nevoid basal cell carcinoma syndrome and non-syndromic keratocystic odontogenic tumors: A case report. [Articolo su rivista]
Ponti, Giovanni; A., Pollio; L., Pastorino; Pellacani, Giovanni; Magnoni, Cristina; S., Nasti; G., Fortuna; Tomasi, Aldo; G. B., Scarrà; Seidenari, Stefania
abstract

Mutations in the Patched homolog 1 (PTCH1) gene lead to an autosomal dominant disorder known as nevoid basal cell carcinoma syndrome (NBCCS) or Gorlin syndrome (GS). Several PTCH1 mutations have been observed in NBCCS associated with keratocystic odontogenic tumors (KCOTs), including non-syndromic KCOTs. The missense mutation c.3277G>C (p.G1093R) in exon 19 of the PTCH1 gene has only been reported in non-syndromic KCOTs. The present study reports for the first time a familial case (father and daughter) of NBCCS and KCOTs, carrying the same c.3277G>C (p.G1093R) germline mutation. This observation suggests that this missense mutation is involved in the pathogenesis of NBCCS as well as in a subset of non-syndromic KCOTs. The identification of a missense mutation may lead to an earlier diagnosis of NBCCS.

2012 - Population-based estimates of the occurrence of multiple vs first primary basal cell carcinomas in 4 European regions [Articolo su rivista]
de Vries, E; Micallef, R; Brewster, Dh; Gibbs, Jh; Flohil, Sc; Saksela, O; Sankila, R; Forrest, Ad; Trakatelli, M; Coebergh, Jw; Proby, Cm; EPIDERM Group (Stockfleth E, Apap C; Fiorentini, Chiara; Magnoni, Cristina
abstract

OBJECTIVE: To estimate the population-based incidence of first and multiple basal cell carcinomas (BCCs) throughout Europe. DESIGN: The registry practices of 4 population-based cancer registries that routinely register BCC incidence were evaluated for inclusion of first and subsequent histologically confirmed BCCs. Where multiple BCCs were not routinely registered, comparisons with hospital databases were made. DATA SOURCES: Cancer registry databases from Finland, Malta, the Netherlands (Eindhoven), and Scotland were inspected for registry of first and multiple BCCs in recent years. Cross-checks with hospital and pathology databases were made to check for completeness. RESULTS: Age-standardized first BCC incidence rates varied between 77 (Malta) and 158 (Eindhoven) per 100 000 person-years. Generally, rates were higher in males than in females, and incidences increased steeply with increasing age. There were approximately 30% more patients with a BCC and 40% to 100% more BCC tumors diagnosed in a given calendar year than were routinely reported for patients with a first primary BCC. The difference between the number of first primary BCCs and the total number of BCCs in a calendar year was generally slightly higher for males than for females and increased substantially with increasing age. CONCLUSION: Currently, routinely reported first BCC incidence rates of the included countries should be multiplied by a factor of 1.3 for an estimate of total number of patients diagnosed as having a BCC in a given year

2012 - Potential impact of interventions resulting in reduced exposure to ultraviolet (UV) radiation (UVA and UVB) on skin cancer incidence in four European countries, 2010-2050 [Articolo su rivista]
de Vries, E; Arnold, M; Altsitsiadis, E; Trakatelli, M; Hinrichs, B; Stockfleth, E; Coebergh, J; Epiderm Group (Ioannides D, Kalabalikis D; Magnoni, Cristina; Fiorentini, Chiara; Majewski, S; Proby, C; Crawford, L.
abstract

BACKGROUND: Behavioural interventions to reduce exposure to ultraviolet radiation (UVR) can reduce risk of skin cancer. OBJECTIVES: To integrate the data and to evaluate the impact of interventions to limit exposure to UVR on skin cancer incidence in four selected countries. METHODS: Using PREVENT, a dynamic simulation model, we modelled the potential for skin cancer prevention in four European countries under various scenarios to avoid damage by UVR. RESULTS: In general, the most effective interventions were those aimed at protecting people during outdoor work and outdoor hobbies against the harmful effects of UVR, and combinations of several interventions. These could in theory lead to reductions of up to 45% in skin cancer cases projected for the year 2050. CONCLUSIONS: The scope for prevention depends on the prevalence of the risk factors in the different countries, as well as the associated risk factors and time lags modelled.

2012 - Risk factors for actinic keratosis in eight European centres: a case-control study [Articolo su rivista]
Traianou, A; Ulrich, M; Apalla, Z; De Vries, E; Bakirtzi, K; Kalabalikis, D; Ferrandiz, L; Ruiz de Casas, A; Moreno Ramirez, D; Sotiriadis, D; Ioannides, D; Aquilina, S; Apap, C; Micallef, R; Scerri, L; Pitkänen, S; Saksela, O; Altsitsiadis, E; Hinrichs, B; Magnoni, Cristina; Fiorentini, Chiara; Majewski, S; Ranki, A; Proby, Cm; Stockfleth, E; Trakatelli, M; Epiderm, Group
abstract

BACKGROUND: There are limited data regarding the association of actinic keratosis (AK) and other types of nonmelanoma skin cancer (NMSC); studies investigating possible correlation of AK with melanocytic naevi are even scarcer. To our knowledge, there are no data examining the risk of AK in people using specific medications. OBJECTIVE: To investigate constitutional and exposure risk factors leading to AK and the coexistence of AK with NMSC and melanoma. METHODS: A multicentre hospital-based case-control study was performed in Finland, Germany, Greece, Italy, Malta, Poland, Scotland and Spain, including 343 patients with actinic keratosis (AK), 409 with squamous cell carcinoma (SCC), 602 with basal cell carcinoma (BCC), 360 with invasive melanoma and 119 with in situ melanoma, and 686 control subjects. Exposures were assessed by questionnaires that were partly self-administered and partly filled out by dermatologists. Unconditional logistic regression modelling was used to assess associations including the influence of phenotypic characteristics, presence of naevi, sun-exposure habits and certain drugs on AK risk. RESULTS: Differences in hair and eye coloration variably influenced the risk for AK, with red hair signifying a seven times higher risk [odds ratio (OR) 6·9, 95% confidence interval (CI) 4·34-11·00), and brown - compared with blue - eyes, about a 40% reduced risk (OR 0·61, 95% CI 0·13-0·92). The darker the skin phototype, the lower the risk for AK, with phototype IV exhibiting nine times less risk of developing AK. Some and many freckles on the arms were associated with an OR of 1·8 (95% CI 1·08-2·81) and 3·0 (95% CI 1·10-3·54), respectively, while overall number of naevi and high educational level were inversely associated with AK. Sun exposure, thiazide diuretics and cardiac drugs had a higher risk for AK. SCC was the most frequent (58%) skin neoplasm coexisting with AKs, followed by BCC (30%), melanoma in situ (12%) and invasive melanoma (6%). CONCLUSION: In this large case-control study from across Europe the expected associations were confirmed for known risk factors. Some possible new risk factors, including cardiac and diuretic drugs, were identified, creating a new field for further investigation in future studies.

2012 - The patient journey: a report of skin cancer care across Europe [Articolo su rivista]
Trakatelli, M; Siskou, S; Proby, C; Tiplica, Gs; Hinrichs, B; Altsitsiadis, E; Kitsou, A; Ferrandiz, L; Aquilina, S; Apap, C; Ulrich, M; Fiorentini, Chiara; Magnoni, Cristina; de Vries, E; Flohil, Sc; Kalokasidis, K; Moreno Ramirez, D; Ruiz de Casas, A; Majewski, S; Ranki, A; Pitkanen, S; Saksela, O; Ioannides, D; Sotiriadis, D; Stockfleth, E; Epiderm, Group
abstract

BACKGROUND: There are poorly documented variations in the journey a skin cancer patient will follow from diagnosis to treatment in the European Union. OBJECTIVES: To investigate the possible difficulties or obstacles that a person with a skin malignancy in the European Union may have to overcome in order to receive adequate medical screening and care for his/her condition. In addition, we wished to explore differences in European health systems, which may lead to health inequalities and health inequities within Europe. METHODS: Ten European countries took part in this investigation (in alphabetical order): Finland, Germany, Greece, Italy, Malta, Poland, Romania, Spain, the Netherlands and the U.K. The individual participants undertook local and national enquiries within their own country and completed a questionnaire. RESULTS: This exercise has identified important differences in the management of a skin cancer patient, reflecting major disparities in health care between European countries. CONCLUSIONS: Further investigation of health disparities and efforts to address health inequalities should lead to improvements in European health care quality and reduction in morbidity from skin cancer.

2012 - Unicystic ameloblastoma associated with the novel K729M PTCH1 mutation in a patient with nevoid basal cell carcinoma (Gorlin) syndrome. [Articolo su rivista]
Ponti, Giovanni; Pollio, A.; Mignogna, M. D.; Pellacani, Giovanni; Pastorino, L.; Bianchi Scarrà, G.; Di Gregorio, C.; Magnoni, Cristina; Azzoni, P.; Greco, M.; Seidenari, S.
abstract

Nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder characterized by a very wide spectrum of clinical signs and symptoms. Here, we report an unusual case of NBCCS in a 38-year-old man with an early onset of clinical signs and symptoms and an associated unicystic ameloblastoma, histopathologically showing basaloid differentiation and intraluminal growth. The odontogenic tumor was surgically enucleated and recurred at the follow-up at 14 months. The proband and his child were identified as gene carriers of the novel K729M PTCH1 missense mutation; other first- and second-degree relatives presented clinical features of NBCCS. Only five other cases of association between ameloblastoma and NBCCS have been reported so far, suggesting that PTCH1 missense mutation might take part in the pathogenesis of keratocystic odontogenic tumors (KCOTs) as well as ameloblastomas.

2012 - Value and prognostic significance of mitotic rate in a retrospective series of pT1 cutaneous malignant melanoma patients [Articolo su rivista]
Ponti, Giovanni; A., Pollio; A. M., Cesinaro; Pellacani, Giovanni; Magnoni, Cristina; Seidenari, Stefania
abstract

INTRODUCTION: Patients affected by thin melanomas (≤1mm) generally have a good prognosis; however, some have a recurrence and eventually die of the disease. The seventh edition of the American Joint Committee on Cancer (AJCC) melanoma staging system, introduced mitotic rate (MR) as one of the primary criteria for staging thin melanoma. MATERIALS AND METHODS: In this study, we sought to determine the prognostic value of mitotic rate in a retrospective cohort of localized primary cutaneous melanoma patients. RESULTS: In total, 286 cases of pT1 primary malignant melanoma occurring in the period 2003-2008 were evaluated. Mitotic counts were re-assessed on standard sections of cases without mitosis and with at least 1 mitosis at diagnosis; 5-year follow-up and recurrence-free survival were available for all patients. Of the 56 radically treated pT1b melanoma patients, 4 (7.1%) had recurrent disease. These data support the efficacy of the incorporation of mitogenicity into AJCC staging for localized cutaneous melanoma and indicate the difficulties in the accuracy and reproducibility of the mitotic count system.

2012 - Variegated Dermoscopy of in situ Melanoma. [Articolo su rivista]
S., Seidenari; S., Bassoli; S., Borsari; C., Ferrari; F., Giusti; Ponti, Giovanni; C., Tomasini; Magnoni, Cristina
abstract

Background: Melanomas in situ (MIS) are difficult to diagnose, lacking well-established dermoscopic descriptors. Objective: The aim of this study was to improve the identification of early melanomas describing the variegated dermoscopic features of MIS and their correlation with demographic and clinical aspects. Methods: Dermoscopic images of 114 histologically proven MIS were evaluated by 3 expert dermoscopists and classified into their main dermoscopic patterns. Dermoscopic features were also considered for their correlation with clinical parameters. Results: Eight different dermoscopic subtypes of MIS were identified: reticular grey-blue (27.2%), reticular (21.1%), multicomponent (20.2%), island (10.5%), spitzoid (7%), inverse network (6.1%), 'net-blue globules' (5.3%) and globular (2.6%). Clinical characteristics of lesions and patients varied according to the different dermoscopic groups. Conclusion: We hypothesize that the different dermoscopic subgroups of MIS correspond to lesions with a different origin and, possibly, various patterns of growth and a different biological behaviour.

2012 - Wave technique for treatment of lower lip cancer [Articolo su rivista]
Salgarelli, Attilio Carlo; Magnoni, Cristina; Bellini, Pierantonio
abstract

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2011 - Cicatricial iatrogenic lower eyelid malposition in skin cancer surgery: results of a combined approach [Articolo su rivista]
Salgarelli, Attilio Carlo; Francomano, Mariangela; Magnoni, Cristina; Bellini, Pierantonio
abstract

Purpose: Malposition of the lower lid, including rounding of the lateral canthal angle, lower eyelidretraction with inferior scleral show, and ectropion, is a relatively frequent complication in the surgicaltreatment of skin cancer of the cheek and zygomatic areas. The tarsal strip technique, in association witha vertical vector cheek lift, is a reliable method for correcting lower lid malposition.Materials and patients: From January 2008 to January 2010, we treated 19 patients with lower eyelidmalposition after skin cancer surgery of the cheek and zygomatic areas. To correct lower eyelidmalposition, we used the tarsal strip technique and a vertical vector cheek lift in all patients.Results: Eleven patients had scleral show and eight patients had ectropion. Sixteen patients obtainedsatisfactory correction of the eyelid malposition in a single surgical procedure, while three patientsrequired a second surgical step to correct the remaining scleral show. Good esthetic and functionalresults were achieved in all cases.Conclusions: The surgical treatment of skin cancer of the cheek and zygomatic areas has the potential forpostoperative sequelae. The tarsal strip technique, in association with a vertical vector cheek lift, isa relatively simple technique for correcting scleral show and ectropion.

2011 - Clinical Study – Reconstruction of Nasal Skin Cancer Defects with Local Flaps [Articolo su rivista]
Salgarelli, Attilio Carlo; Bellini, Pierantonio; Multinu, A.; Magnoni, Cristina; Francomano, M.; Fantini, F.; Consolo, Ugo; Seidenari, S.
abstract

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2011 - Reconstruction of nasal skin cancer defects with local flaps. [Articolo su rivista]
Salgarelli, Attilio Carlo; Bellini, Pierantonio; A., Multinu; Magnoni, Cristina; M., Francomano; F., Fantini; Consolo, Ugo; Seidenari, Stefania
abstract

Reconstruction of nasal defects must preserve the integrity of complex facial functions and expressions, as well as facial symmetry and a pleasing aesthetic outcome. The reconstructive modality of choice will depend largely on the location, size, and depth of the surgical defect. Individualized therapy is the best course, and numerous flaps have been designed to provide coverage of a variety of nasal-specific defects. We describe our experience in the aesthetic reconstruction of nasal skin defects following oncological surgery. The use of different local flaps for nasal skin cancer defects is reported in 286 patients. Complications in this series were one partial flap dehiscence that healed by secondary intention, two forehead flaps, and one bilobed flap with minimal rim necrosis that resulted in an irregular scar requiring revision. Aesthetic results were deemed satisfactory by all patients and the operating surgeons. The color and texture matches were aesthetically good, and the nasal contour was distinct in all patients. All scars were inconspicuous and symmetrical. No patient had tenting or a flat nose.

2011 - Synergistic Use of Local Flaps for Total Lower Lip Reconstruction [Articolo su rivista]
Salgarelli, Attilio Carlo; Bellini, Pierantonio; Magnoni, Cristina; Anesi, Alexandre; Collini, M.
abstract

The loss of tissue in the lower lip has beentreated using different techniques dependingon the size and location (median or lateral) of thedefect.1,2 Plastic surgery for small defects of up totwo-thirds of the lower lip relies on surgicaltechniques such as direct suturing of the resectionmargins with or without Z-plasty. The methodsthat Gillies and Millard,3 Estlander,4 Abbe´,5 andKarapandzic6 proposed, as well as the staircasetechnique,7 have been used for reconstructionsinvolving up to two-thirds of the lip, whereas theproximity of the flaps is suitable for treatingcomplete lower lip losses, according to Von Bruns,8Bernard,9 and Bernard-Freeman-Fries.10–12The multiplicity of techniques reported in the literatureindicates that there is no ideal method fortotal lower lip reconstruction. It is generally agreedthat reconstruction after resection of more thantwo-thirds of the lower lip produces visible scarsand alters the kinetics of smiling.1,2,7The Sabattini-Abbe´ flap is a full-thickness one-lipflap that is pedicled on the superior or inferiorlabial vessels and rotated 180° to fit into the defectof the opposite lip. The Sabattini-Abbe´ flap can beused for reconstructing up to two-thirds of the lip.The staircase technique, used to close full-thicknessdefects of up to two-thirds of the lower lip, is basedon the advancement flap. Johanson and colleaguesfirst described it in 1974,13 and by Dado andAngelats,14 Stiernberg,15 Kuttenberger and Hardt16and Salgarelli and colleagues7 subsequentlymodified it. Reconstruction of the lower lip isperformed using a lateral advancement flap fromthe lateral parts of the lip and chin, in a stepwisefashion.7Here, we propose the synergistic use of thestaircase technique and a Sabattini-Abbe´ flap forreconstructions involving more than two-thirds ofthe lower lip (Figures 1A and B).

2011 - 18beta-glycyrrhetinic acid and glabridin prevent oxidative DNA fragmentation in UVB-irradiated human keratinocyte cultures. [Articolo su rivista]
Veratti, Eugenia; Rossi, Tiziana; Giudice, Stefania; Benassi, Luisa; Bertazzoni, Giorgia; Morini, D; Azzoni, Paola; Bruni, E; Giannetti, Alberto; Magnoni, Cristina
abstract

To investigate on the possible protective activity of glycyrrhizin (GL), 18β-glycyrrhetinic acid (18β-GA) and glabridin (GLB) against UVB radiation damage in human keratinocyte cultures the following assays were performed: MTT, Comet, ROS generation and Western blot test (for the p53 activation, BCL-2 regulation and PARP cleavage). Results: 18β-GA and GLB prevented direct and indirect DNA fragmentation avoiding apoptosis activation.in human keratinocytes.

2010 - Skin Squared [Spin Off]
Pellegrini, Massimo; Giannetti, Alberto; Magnoni, Cristina; Benassi, Luisa; Areta International, s. r. l.; Università di Modena e., Reggio Emilia
abstract

2009 - In vitro evaluation on the anti-apoptotic and anti-oxidant effect of a series of natural polyphenols on UVB- irradiated normal human keratinocytes and melanocytes [Abstract in Atti di Convegno]
E., Bruni; Giudice, Stefania; Veratti, Eugenia; Magnoni, Cristina; Benassi, Luisa; Pellati, Federica; Benvenuti, Stefania; Rossi, Tiziana
abstract

UV radiation, in particular its UVB component, is an important environmental factor in the pathogenesis of skin aging and cancer.One of the features of UVB-caused DNA damage is the formation of cyclobutane pyrimidine dimers and (6-4) photoproducts. Further, indirect DNA damage is also caused by increase in the level of reactive oxygen species (ROS) (1) that cause oxidative damage reacting with DNA, proteins, fatty acids and saccharides (2). Such injuries result in a number of harmful effects: disturbed cell metabolism, morphological and ultrastructural changes, attack on the regulation pathways and, alterations in the differentiation, proliferation and apoptosis of skin cells (3). These processes can lead to photoaging and skin cancer development. In order to avoid UVB radiation damage, phytocompounds and antioxidants as photoprotectives has been considered (4). In recent years naturally occurring herbal compounds such as phenolic acids, flavonoids, and high molecular weight polyphenols have gained remarkable attention as strong protective agents (5,6). In the present study we considered a series of natural polyphenols with the aim to investigate on their anti-oxidant and anti-apoptotic effect in UVB-irradiated normal human keratinocytes and melanocytes.Methods: growing concentration (from 10 to 100 M) of forty-six polyphenols selected by the Department of Pharmaceutical Sciences of the University of Modena were added to normal human keratinocytes and melanocytes coltures. In the first part of the study we assessed the activity on UVB-irradiated normal human keratinocytes and melanocytes. These substances were submitted to a primary in vitro screening by MTT test in order to valuate proliferation rate. Subsequently, 2’7’- dichlorodihydrofluorescein diacetate (DCF) assay was performed to determine the formation of intracellular ROS. The results showed that glabridin, 18β-glycyrrhetinic acid, kaempferol, quercetin, rutin and butein presented interesting properties and a low toxicity. In the second part of the study the antiapoptotic effects of these six selected compounds in UVB-irradiated (50 mJ/cm2/11’and 44’’) normal human keratinocytes and melanocytes was investigated by the western blot analysis.Results: with regard to analysed polyphenols, the results showed that a pre-treatment of human keratinocytes and melanocytes with these molecules inhibited UVB mediated apoptosis through the activation of p53, down-regulation of bcl-2 and bid full-length, up- regulation of bax and inhibition of PARP cleavage. For these purpose we proposed that some substances could be combined to evaluate their synergistic effects.Conclusion: To further elucidate the molecular mechanism of polyphenol or more generally phytocompounds mediated apoptosis, we decided to investigate cell cycle analysis and the extrinsic and intrinsic apoptotic pathways. Human skin is constantly exposed to the UV radiation present in sunlight. This may induce a number of pathobiological cellular changes. The development of novel preventive and therapeutic strategies depends on our understanding of the molecular mechanism of UV-damage. Phytochemical that were identified may be candidates for prevention of adverse effects of UV radiation and melanoma on the skin and evaluation of there clinical efficacy is awaited.

2009 - Multidisciplinary chronic leg ulcers service in Modena: data anaysis. [Articolo su rivista]
Lonardi, Roberto; Curti, Tiziano; F., Neri; D., Olezzi; Giannetti, Alberto; Magnoni, Cristina
abstract

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2007 - A new role of phase I and phase II enzyme in keratinocytes UVB induced apoptosis. [Abstract in Rivista]
Benassi, Luisa; Bertazzoni, Giorgia; Giudice, Stefania; Giannetti, Alberto; Magnoni, Cristina
abstract

A new role of Phase I and Phase II enzymes in keratinocytes UVB induced apoptosis.UVB is the major cause for cutaneous malignancies in the human population. The skin is able to activate anti-oxidants and enzymatic detoxification reactions to neutralize reactive photochemical products. Apoptosis, removing irreversibly DNA-damaged and potentially neoplastic cells, represents a major defence mechanism towards malignant transformation. Very little is known about the role of cutaneous cytochrome P450 (CYP450) isoenzymes and glutathione S-transferase (GST) in UVB response. Although not yet fully investigated, it has been studied a possible relationship between CYP450 inductors and UVB initiated apoptosis in rat hepatocytes. Altered high levels of GST has been directly correlated to resistance to chemotherapeutic drugs suggesting that GST plays a role in prevention of apoptosis. Our study is focused on phase I and phase II enzyme activities in normal human keratinocytes. In the first part of the study we demonstrated that CYP450 (1A1 and 2B1) and GST are induced not only by classical inducers such as β-naphthoflavone, 3-methylcholanthrene, phenobarbital but also by UVB radiations (50 mJ/cm2). Differentiated keratinocytes were employed for all the experiments as confirmed by immunoblotting with Cytokeratin 10, a specific marker of the basal spinous transition. In the second part we evaluated a possible involvement of these enzyme in UVB mediated apoptosis process. Western blot analysis of Bcl2 expression and PARP cleavage showed that inhibition of CYP450 by Proadifen prevented UVB induced apoptotic cell death. In contrast, the diuretic drug, ethacrinic acid, a GST inhibitor, was able to improve UVB induced apoptosis. In order to confirm the anti-apoptotic activity displayed by GST, when GST activity was increased by Phenobarbital, UVB apoptosis was prevented.These results suggest that UVB radiations may play a critical role as tumour promoters even through the regulation of CYP450 and GST metabolising enzymes.

2007 - New thymidylate synthase inhibitors induce apoptosis in melanoma cell lines [Articolo su rivista]
Giudice, Stefania; Benassi, Luisa; Bertazzoni, Giorgia; Costi, Maria Paola; A., Gelain; Venturelli, Alberto; Bernardi, Chiara; Gualdi, Giulio; A., Coppi; Rossi, Tiziana; Giannetti, Alberto; Magnoni, Cristina
abstract

Malignant melanoma is particularly resistant to conventional chemotherapy and radiotherapy. For this reason in the past years a huge variety of new compounds has been developed with potential chemotherapeutic activity which needs to be tested in vitro and in vivo. We investigated the in vitro action of three new experimental antifolate substances (MR7, MR21 and MR36) with a critical target for thymidylate synthase (TS), an essential enzyme for DNA synthesis. The response of two melanoma cell lines (SK-MEL-2 derived from malignant melanoma metastasis and SK-MEL-28 derived from primary malignant melanoma) was examined after treatment with these substances. The antifolate agents induced apoptosis in SK-MEL-2 and SK-MEL-28 cells as confirmed by the TUNEL technique and Comet Assay. Western-blot analysis showed a down-regulation of Bcl-2 protein level and PARP cleavage, otherwise p53 and Bax expressions were not modulated. Moreover, these antifolate-induced apoptosis was accompanied by both pro-caspase-9 and -8 activations. These results were supported by the use of the pan-caspases inhibitor Z-VAD-FMK that almost completely decreased the amount of apoptosis in both the melanoma cell lines treated with antifolate. In conclusion our results show that TS inhibitors are able to induce apoptosis through a caspase-mediated pathway, but without the involvement of the p53/Bax signalling. (c) 2006 Elsevier Ltd. All rights reserved.

2007 - Transcriptional profiles in melanocytes from clinically unaffected skin distinguish the neoplastic growth pattern in patients with melanoma [Articolo su rivista]
Magnoni, Cristina; Tenedini, Elena; Ferrari, Francesco; Benassi, Luisa; Bernardi, Chiara; Gualdi, Giulio; Bertazzoni, Giorgia; Roncaglia, Enrica; Fantoni, Luca Isaia; Manfredini, Rossella; Bicciato, Silvio; Ferrari, Sergio; Giannetti, Alberto; Tagliafico, Enrico
abstract

Background It is generally accepted that sunlight may contribute to the development of melanoma. Objectives To analyse gene expression of melanocytes obtained from clinically unaffected skin of patients with melanoma and healthy controls before and after exposure to ultraviolet B radiation. Methods Using GeneChip array technology, the gene expression of melanocytes obtained from the two donor groups was profiled, in order to identify transcriptional differences affecting susceptibility to melanoma. Results The data collected did not show any difference between the expression profiles of melanocytes purified from normal donors and from patients with melanoma that was able to give a statistically significant class separation. However, by means of unsupervised clustering our data could be divided into two main classes. The first class included the transcriptome profiles of melanocytes obtained from skin samples of patients with a vertical growth phase (VGP) melanoma, while the second class included the transcriptome profiles of melanocytes obtained from skin samples of patients with a radial growth phase (RGP) melanoma. Conclusions These data suggest that melanocytes in patients with VGP and RGP melanomas show significant differences in gene expression profiles, which allow us to classify patients with melanoma also from clinically unaffected skin.

2006 - Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells. [Articolo su rivista]
Mavilio, Fulvio; Pellegrini, Graziella; Ferrari, S.; DI NUNZIO, Francesca; Di Iorio, E.; Recchia, Alessandra; Maruggi, Giulietta; Ferrari, G.; Provasi, E.; Bonini, C.; Capurro, S.; Conti, A.; Magnoni, Cristina; Giannetti, Alberto; DE LUCA, Michele
abstract

The continuous renewal of human epidermis is sustained by stem cells contained in the epidermal basal layer and in hair follicles. Cultured keratinocyte stem cells, known as holoclones, generate sheets of epithelium used to restore severe skin, mucosal and corneal defects. Mutations in genes encoding the basement membrane component laminin 5 (LAM5) cause junctional epidermolysis bullosa (JEB), a devastating and often fatal skin adhesion disorder. Epidermal stem cells from an adult patient affected by LAM5-beta3-deficient JEB were transduced with a retroviral vector expressing LAMB3 cDNA (encoding LAM5-beta3), and used to prepare genetically corrected cultured epidermal grafts. Nine grafts were transplanted onto surgically prepared regions of the patient's legs. Engraftment was complete after 8 d. Synthesis and proper assembly of normal levels of functional LAM5 were observed, together with the development of a firmly adherent epidermis that remained stable for the duration of the follow-up (1 year) in the absence of blisters, infections, inflammation or immune response. Retroviral integration site analysis indicated that the regenerated epidermis is maintained by a defined repertoire of transduced stem cells. These data show that ex vivo gene therapy of JEB is feasible and leads to full functional correction of the disease.

2006 - CYP450 and GST are induced by xenobiotics in normal human keratinocytes and play a different role in UVB-mediated apoptosis. [Abstract in Rivista]
Benassi, Luisa; Magnoni, Cristina; Bertazzoni, Giorgia; Giudice, Stefania; Gualdi, Giulio; Bernardi, Chiara; Seidenari, Stefania; Giannetti, Alberto
abstract

Skin enzymatic detoxification systems may provide a barrier for protection against the external environment. UVB is the major cause for cutaneous malignancies in the human population. The skin is able to activate anti-oxidants and enzymatic detoxification reactions to neutralize reactive photochemical products. Very little is known about the behaviour of cutaneous metabolizing enzymes after ultraviolet B exposure. In the first part of the present study we analysed the induction and expression of cytocrome P450 (CYP450) phase I and gluthatione S transferase (GST) phase II enzymes in normal human keratinocytes in culture after exposure to classical inducers such as: β-naphthoflavone (BNF), 3-methylcholanthrene (MC), phenobarbital (PB) and also to ultraviolet B (UVB) radiation. Phase I enzymes (CYP450 1A1, 1A2, 1B2 and 2B1, 2B2) were investigated through 7-ethoxyresorufìn O-deethylase (EROD) and 7-pentoxyresorufìn O-depenthylase activities (PROD), while GST was analysed throught the coniugation of 1-chloro-2,4 dinitrobenzene with reduced gluthatione.MC, BNF and UVB exposure resulted in a dose-dependent and time dependent induction of CYP450 confirmed by immunoblotting assay. GST was induced by PB and inhibited by UVB. According to these results in the second part of this study we analysed the role of these enzymes in UVB induced apoptotic process. Apoptosis is a key mechanism in the cellular homeostasis and can be triggered by exogenous insults and in particular by genotoxic agents. The exact function of these enzymes in carcinogenesis remains unclear. Western blot analysis of Bcl2 expression showed that inhibition of CYP450 prevented UVB induced apoptotic cell death. On the contrary inhibition of GST, by ethacrynic acid, showed to improve UVB induced apoptosis. These experimental findings stress the value that CYP450 and GST enzymes may play an important role in keratinocytes apoptosis.

2006 - Dermoscopic patterns as predictors of sentinel lymph node metastasization in cutaneous melanoma [Articolo su rivista]
M., Curci; Pellacani, Giovanni; Magnoni, Cristina; F., Fantini; A., Cimitan; Seidenari, Stefania
abstract

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2006 - New Thymidylate synthase inhibitor induces apoptosis in melanoma cell lines. [Abstract in Atti di Convegno]
Giudice, Stefania; Benassi, Luisa; Magnoni, Cristina; Bertazzoni, Giorgia; Costi, Maria Paola; Rossi, Tiziana; Bernardi, Chiara; Gualdi, Giulio; Giannetti, Alberto
abstract

Malignant melanoma is an aggressive disease and its rapidly increasing world-wide. This type of tumor is particularly resistant to conventional chemotherapy and radiotherapy. For this reason in the last years has been developed a variety of a new chemotherapeutic agents. We investigated the action in vitro of four experimental antifolate substances (MR7, MR2, MR36 and NMR707) with a critical target for thymidylate synthase (TS), an essential enzyme for DNA synthesis. These compounds behave as antifolate but show a new structure with respect to all know antifolato agents. We studied how these substances may influence apoptosis in melanoma cell lines SK-MEL-2 (derived from malignant melanoma metastasis) and SK-MEL-28 (derived from primary malignant melanoma).The antifolate agents induced apoptosis in SK-MEL-2 and SK-MEL-28 cells as confirmed by TUNEL technique and Comet Assay. Western-blot analysis showed down regulation of Bcl-2 protein level, up-regulation of p21 and Bax protein level and PARP cleavage especially for MR36, MR7 and MR21. Moreover, these antifolate-induced apoptosis was accompanied by caspase-9 and -8 activation. In addition, a different sensibility for apoptosis was observed for the two cell lines. In fact, in primary melanoma cells, apoptosis occurred also at low concentrations especially for MR7 and MR36. Our results show that TS inhibitors are able to induce apoptosis and for this reason may play a potential role as new therapeutic agents for melanoma, but further studies are necessary to understand their molecular mechanism.

2006 - Pharmacological and toxicological evaluation of a new series of thymidylate synthase inhibitors as anticancer agents [Articolo su rivista]
Benassi, Luisa; Magnoni, Cristina; Giudice, Stefania; Bertazzoni, Giorgia; Costi, Maria Paola; Rinaldi, Marcella; Venturelli, Alberto; A., Coppi; Rossi, Tiziana
abstract

Thymidylate synthase (TS) is responsible for catalysing the de novo biosynthesis of doexythymidine monophosphate and is a target for many anticancer drugs. A series of thymidylate synthase inhibitors (TSIs), synthesised in our laboratory, were submitted to primary anticancer screening by the National Cancer Institute (NCI). Four compounds, 3,3bis(4-methoxyphenyl)-1H, 3H-naphtho[1,8-cd]pyran-1-one (MR7), 6-chloro-3,3-bis(4-hydroxyphenyl)-]H,3H-naphtho[1,8cdjpyran-1-one (MR21), 3,3-bis(3-fluoro-4-hydroxyphenyl)IH,3H-naphtho[1,8-cd]pyran-l-one (MR35) and 6-bromo-3,3bis(3-chloro-4-hydro.xyphenyl)-1H,3H-naphtho[1,8-cd]pyran-l- one (MR36), passed the criteria and were automatically scheduled for evaluation against the full panel of 60 human tumour cell lines. In this study, the antiproliferative activity of the substances against SK-MEL-2 cells (from metastatic tissue) and SK-MEL-28 cells (from primary malignant melanoma cells) was investigated. Neutral Red uptake and the MTT test were performed to confirm the results of the NCI, and [H-3]thymidine incorporation was performed as a test of the proliferation rate. Our results indicated that compounds MR21 and MR36 were the most active agents and the [H-3]-thymidine test was the best in predicting toxicity against melanoma cells.

2005 - Effects of glycyrrhizin on UVB-irradiated melanoma cells [Articolo su rivista]
Rossi, Tiziana; Benassi, Luisa; Magnoni, Cristina; Ruberto, Ippazio Antonio; A., Coppi; Baggio, Giosuè Gabriele
abstract

It is known that liquorice root is rich in compounds which exert several pharmacological actions. In the present study, we evaluated the effect of glycyrrhizin (the main constituent of liquorice root) and of its metabolite aglycone, 18 beta-glycyrrhetinic acid, on UVB-irradiated human melanoma cells: SKMEL-2 from metastatic tissue and SKMEL-28 from primary malignant melanoma. Tests performed (Trypan blue exclusion test, MTT and Western blot) showed that glycyrrhizin is not toxic for both types of cells. In SKMEL-28 cells, Bcl-2 expression was low after UVB irradiation, but it was increased when treated with glycyrrhizin. On the contrary, in the SKMEL-2 cell culture, Bcl-2 expression was not modified by the substances under study. The results show that g cyrrhizin treatment might offer protection from the damage induced in humans by UVB radiation, while it seems to be ineffective on metastatic cells. Further studies must be performed to understand the mechanism of the protective effect.

2005 - TSI INDUCES APOPTOSIS WITH A P53 INDIPENDENT PATHWAY IN MELANOMA CELL LINES AND IN NORMAL HUMAN MELANOCITES [Abstract in Rivista]
Benassi, Luisa; Magnoni, Cristina; Bertazzoni, Giorgia; Giudice, Stefania; Costi, Maria Paola; Rossi, Tiziana; Bernardi, Chiara; Gualdi, Giulio; Giannetti, Alberto
abstract

Melanoma patients have a very poor prognosis and this type of tumor is particular resistant toconventional chemotherapy and radiotherapy. For this reason in the last years has beendeveloped a variety of new therapeutic agents. We investigated four experimental antifolatesubstances (MR7, MR36, MR21 and NMR707) with a critical target for thymidylate synthase(TS), an essential enzyme for DNA synthesis and repair. All the compounds were tested fortheir inhibitory properties against TS enzymes from different species: Lactobacillus casei,Pneumocystis carinii, Cryptococcus neoformans and human. These compounds behave asantifolates but show new structure with respect to all known antifolate agents. We studiedhow these substances may influence apoptosis in melanoma cell lines SKmel 2 (derived frommalignant melanoma metastasis) and SKmel 28 (derived from primary malignant melanoma)and normal human melanocytes. The antifolate agents induced apoptosis in Skmel 2 andSkmel 28 cells. These results were confirmed by Comet Assay. Western blot analysis showeddown regulation of the level of Bcl2 and PARP clivage especially for MR36, MR7 and MR21.Moreover a different apoptotic behaviour for the two cell lines was observed, in fact inprimary melanoma cells apoptosis occurred also at low concentrations especially for MR7.After chemotherapeutic treatment with all the tested compounds p53 was not modulatedindicating a p53 independent pathway for apoptosis. Caspases 8 and 9 clearly mediateantifolate agents apoptosis independent of the status of p53. Our results show that TSinhibitors inducing apoptosis may play a future potential role as new therapeutic agents formelanoma, but further studies are necessary to understand their molecular mechanism.

2004 - Apoptosis: Escaping strategies in human skin cancer (Review) [Articolo su rivista]
Giannetti, Luca; Consolo, Ugo; Magnoni, Cristina; Lorenzo Lo, Muzio
abstract

Apoptosis must be considered a rescue mechanism that controls the integrity of the cell erasing aberrant clones and it is likely that failure of apoptosis constitutes a key factor responsible for tumour formation, progression and resistance to drugs. Several genes (e.g. survivin, bcl-2 family, p53) are involved in these pathways. Much work still needs to be done to identify the molecular pathway of apoptosis resistance but restoration of the apoptotic response represents a crucial new therapeutic target for cancer treatment.

2004 - Induction of phase I and phase II enzyme activities by UVB and xenobiotics in normal human keratinocytes. [Abstract in Rivista]
Benassi, Luisa; Magnoni, Cristina; Bertazzoni, Giorgia; Caselli, Monica; Seidenari, Stefania; Giannetti, Alberto
abstract

The function of the skin is to provide a barrier for protection against the external environment. Relatively little is known about the overall role of CYP450 in the metabolism of xenobiotics or endogenous cellular compounds in the skin. The aim of this study was to analyse the expression and induction of several drugs metabolizing enzymes. To this purpose both phase I and phase II enzymes activities were studied in normal human keratinocytes in culture after exposure to UVB radiation and to classical cytochrome inducers: β-naphthoflavone (BNF), 3-methylcholanthrene (MC), phenobarbital (PB). We investigated 7-ethoxyresorufìn O-deethylase (EROD) and 7-pentoxyresorufìn O-depenthylase activities (PROD). Normal human keratinocytes were cultured with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12. At confluence cells were incubated with inducers or irradiated with different doses of UVB. The microsomal fraction was studied by western-blot analysis. The MC-induced EROD activity was up to fourfold higher when compared with BNF induced activity. UVB exposure resulted in a dose-dependent (25-75 mJ) and time dependent (6-24 h) induction of CYP450 1A1. Immunoblotting assay showed expression for CYP450 1B1 not only keratinocytes but also in melanocytes. Proadifen, an inhibitor of CYP450-monooxygenase, led to a significant decrease in EROD activity. The results of the present study clearly show that irradiation with UVB is capable of modifying the activity of CYP450 isoenzymes in keratinocytes. The phase II enzyme gluthatione S transferase activity (GST) was induced by UVB and PB. These experimental findings stress the value of epidermal cell culture for pharmaco-toxicological studies of topical agents used in dermatology.

2003 - Decrease in toxic potential of mixed tensides maintained below the critical micelle concentration: An in vitro study [Articolo su rivista]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; Rinaldi, M; Fontanesi, Claudio; Seidenari, Stefania
abstract

Sodium lauryl sulphate (SLS) is an anionic tenside widely utilized in commercial topical preparations that may cause skin irritation. It has been shown that the barrier damage caused by SLS in vivo is lower when SLS is used in combination with other tensides which are able to reduce the critical micelle concentration (CIVIC). The aim of our study was to evaluate if the cytotoxic effect of SLS is reduced by the association with different tensides also at concentrations well below the CIVIC. Normal human keratinocytes from plastic surgery were grown in serum-free medium. At subconfluency, the cells were treated with SLS at a dose of 0.0025% in combination with cocamidopropyl betaine, Tween 20 and Tween 80 at the minimum toxic dose. Following tenside treatment, the culture medium was changed, and after 24 h the cells were collected for H-3-thymidine incorporation, the MTT (3-[4,5-dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay and neutral red (NR) uptake. The cytotoxic effect on normal human keratinocytes, as evaluated by 3H-thymidine incorporation, MTT assay and NR uptake, was significantly decreased by the combination with all the tested tensides. The correlation between cytotoxicity and physical properties was also studied by a conductimetric assay to investigate the mechanism involved in this toxicity reduction. Copyright

2003 - Induction of cytochrome P450 enzyme activity by UVB and xenobiotics in normal human keratinocites. [Abstract in Rivista]
Bertazzoni, Giorgia; Benassi, Luisa; Magnoni, Cristina; Caselli, Monica
abstract

The function of the skin is to provide a barrier for protection against the external environment. Relatively little is known about the overall role of CYP450 in the metabolism of xenobiotics or endogenous cellular compounds in the skin. The aim of this study was to analyse the expression and induction of several drugs metabolizing enzymes involved in either phase I or phase II reactions, in proliferanting human keratinocytes after exposure to UVB radiation and to three classical cytochrome inducers such as: β-naphthoflavone (BNF), 3-methylcholanthrene (MC), phenobarbital (PB). We investigated 7-ethoxyresorufìn O-deethylase (EROD) and 7-pentoxyresorufìn O-depenthylase activities (PROD). Normal human keratinocytes were cultured with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12. At confluency cells were incubated with inducers or irradiated with different doses of UVB. The microsomal fraction was studied by western-blot analysis. The MC-induced EROD activity was up to 4 fold higher when compared with BNF induced activity. UVB exposure resulted in a dose-dependent (25-75 mJ) and time dependent (6-24 h) induction of CYP450 1A1. Immunoblotting assay showed expression for CYP450 1B1 for both keratinocytes and melanocytes. Proadifen, an inhibitor of CYP450-monooxygenase, led to a significant decrease in EROD activity. The results of the present study clearly show that irradiation with UVB is capable of modifying the activity of CYP450 isoenzymes in keratinocytes. The phase II enzyme gluthatione S transferase activity (GST) was induced by UVB and PB. These experimental findings stress the value of epidermal cell culture for pharmaco-toxicological studies of topical agents used in dermatology.

2003 - Induzione dell'attività degli enzimi di fase I e II nei cheratinociti umani normali. [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; Caselli, Monica; Seidenari, Stefania
abstract

La cute può essere considerata come un effettivo distretto di biotrasformazione di composti chimici. Lo scopo di questo lavoro è stato analizzare l'espressione dei sistemi enzimatici per il metabolismo di fase I e fase II nei cheratinociti umani in proliferazione dopo irradiazione con raggi UVB e dopo esposizione a tre classici induttori: beta-naphthoflavone (BNF), 3-methylcholantrene (MC), phenobarbital (PB). Cheratinociti umani normali sono stati coltivati con fibroblasti 3T3 mitomicinati in Dulbecco's modified Eagle's medium/Ham's F12. A subconfluenza le cellule sono state incubate con gli induttori o irradiate a differenti dosi di UVB. La fase I è stata analizzata con attività 7-ethoxyresorufin O-deethylase (EROD) e 7-pentoxyresorufin O-depenthylase (PROD). la frazione microsomiale è stata studiata mediante analisi western blot. L'attività EROD indotta dall'MC era più alta di 4 volte rispetto al BNF. L'esposizione a raggi UVB risultava dose dipendente (50-75 mJ) e tempo dipendente (6-24h). L'attività della glutatione-S-transferasi(GST), enzima di fase II determinato con cinetica enzimatica, veniva significativamente espressa con UVB e PB. Questi risultati dimostrano il ruolo della cute nel metabolismo esogeno e la possibilità di utilizzare colture di cellule epidermiche per studi farmacotossicologici di agenti topici usati in dermatologia.

2002 - Induction of cytochrome P450 enzyme activity by UVB and xenobiotics in normal human keratinocytes and melanocytes [Abstract in Rivista]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; Caselli, Monica; Seidenari, Stefania
abstract

Cytochrome P450 (CYP450) play a major role in the bioactivation of procarcinogenesis in target tissue and the expression of this enzyme i san important determinant of human susceptibility to cancer. Relatively little is known about the overall role of CYP450 in the metabolism of xenobiotics or endogenous cellular compounds in the skin. The aim of this study was to analyse the expression of cytochrome enzymes in proliferanting human keratinocytes and melanocytes after exposure to UVB radiation and to three classical cytochrome inducers such as: β-naphthoflavone (BNF), 3-methylcholanthrene (MC), phenobarbital (PB). We investigated 7-ethoxyresorufìn O-deethylase (EROD) (which is CYP450 1A1 dependent) and 7-pentoxyresorufìn O-depenthylase activities (PROD) (CYP450 2B1 dependent)activities. Normal human keratinocytes were cultured with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12 or with KGM serum-free medium. Melanocytes were grown in medium 154. At confluency cells were incubated with inducers or irradiated with different doses of UVB. At different times after treatments, cells were harvested for in vitro measurement of CYP450 induction. The microsomal fraction was studied by western-blot analysis. Low, but measurable levels of CYP activity were detected in both basal and differentianting keratinocytes. The MC-induced EROD activity was up to 4 fold higher when compared with BNF induced activity. UVB exposure resulted in a dose-dependent (10-75 mJ) and time dependent (4-24 h) induction of CYP450 1A1 for keratinocytes and CYP450 2B1 for melanocytes. Immunoblotting assay showed expression for CYP450 1B1 for both keratinocytes and melanocytes. Proadifen, an inhibitor of CYP450-monooxygenase, led to a significant decrease in EROD activity. The results of the present study clearly show that irradiation with UVB is capable of modifying the activity of CYP450 isoenzymes not only in keratinocytes but also in melanocytes.These experimental findings stress the value of epidermal cell culture for pharmaco-toxicological studies of topical agents used in dermatology.

2002 - Neutral sphingomyelinase is modulated by glutathione in keratinocytes UVB-induced apoptosis [Abstract in Rivista]
Magnoni, Cristina; Benassi, Luisa; Rinaldi, Mr; Bertazzoni, Giorgia; Giannetti, Alberto
abstract

The sphingomyelin pathway is an ubiquitous, evolutionary conserved signaling system which plays a role in intracellular signal transduction. The action of a ligand binding to a surface receptor results in early activation of the enzyme sphingomyelinase (SMases) with consequent hydrolysis of membrane sphingomyelin (SM) and generation of ceramide. Increase in intracellular ceramide level is followed by three major cellular responses: cell growth arrest, induction of cell differentiation and/or induction of apoptosis. Recent evidences have shown that reduced glutathione (GSH), but not other antioxidative agent, is able to inhibit apoptotic cell death and necrosis induced by hypoxia in PC12 cells. This protective effect is mediated by GSH direct inhibition of neutral SMase activity and ceramide formation. In the present study we report that GSH inhibits activation of neutral SMases and generation of ceramide and partially prevents apoptotic cell death induced by UVB radiation in keratinocytes. Normal human keratinocytes were cultivated with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12 medium. At preconfluency cells were incubated with GSH for 2h and then irradiated with a UVB dose of 75mJ per cm2. At different times after UVB irradiation, cells were harvested for in vitro measurement of neutral SMaese activity, lipid extraction and Western blot analysis. In vitro measurement of neutral SMase activity showed an early induction 15 min after exposure with a subsequent decrease to control level after 2h. Exposure to UVB radiation resulted in a rapid sphingomyelin hydrolysis and generation of ceramide as measured by TLC analysis. The ceramide accumulation started at 15 min after UV exposure and progressively increased up to 24h. Addition of GSH significantly inhibited activation of neutral SMase and generation of ceramide in UVB-treated keratinocytes. Moreover UVB-induced cleavage of PARP, a marker of the apoptotic response, was partially inhibited by GSH treatment. This data indicate that GSH plays a critical role in UVB signaling pathway regulating neutral SMase activity.

2002 - Ultraviolet B radiation induces activation of neutral and acidic sphingomyelinases and ceramide generation in cultured normal human keratinocytes [Articolo su rivista]
Magnoni, Cristina; Euclidi, Emanuela; Benassi, Luisa; Bertazzoni, Giorgia; Cossarizza, Andrea; Seidenari, Stefania; Giannetti, Alberto
abstract

The sphingomyelin pathway is an ubiquitous, evolutionary conserved signaling system which transduces an extracellular signal into the cell. During the past few years increasing evidence has shown that the sphingolipid ceramide may play a role as a second messenger in intracellular signal transduction. The ceramide generation via sphingomyelinase (SMase) is followed by three major cellular responses: cell growth arrest, induction of cell differentiation and/or induction of programmed cell death or apoptosis. The aim of this study is to investigate whether activation of SMases and generation of ceramide can be induced by UVB radiation in normal human keratinocytes. The present data show that exposure to UVB radiation results in rapid generation of ceramide. The ceramide accumulation starts 15 min after UV exposure and progressively increases up to 24 h. In vitro measurement of SMase activity following exposure to UVB evidences an activation of both neutral and acidic SMases. Moreover, UVB induces apoptosis in normal human keratinocytes as shown by TUNEL technique and FACS analysis. These data indicate that UVB induced ceramide generation and activation of both neutral and acidic SMases, suggesting that sphingolipids metabolism may be involved in the UVB signaling pathway. (C) 2002 Elsevier Science Ltd. All rights reserved.

2001 - Assesment of the cytotoxicity of experimantal calcium releasing cements. [Abstract in Rivista]
Gatti, Antonietta; R. Z., Legeros; Monari, Emanuela; S., Lin; Magnoni, Cristina; Cazzella, Francesco
abstract

The purpose of this study is to evaluate the biocompatibility in rabbit of calcium phosphates. The materials differd in the degree of biodegradation and can therefore be used for application requiring rapid or slow biodegradation.

2001 - Il glutatione endogeno regola l'attivazione della n-smase nel processo di apoptosi indotta dai raggi UVB nei cheratinociti umani normali in coltura. [Abstract in Atti di Convegno]
Magnoni, Cristina; Euclidi, Emanuela; Bertazzoni, Giorgia; Benassi, Luisa; Vaschieri, Cristina; Seidenari, Stefania; Giannetti, Alberto
abstract

I raggi UVB sono le radiazioni maggiormente responsabili degli effetti fotobiologici a livello cutaneo. Come è noto dalla letteratura, i raggi UVB inducono apoptosi nei cheratinociti umani normali, ma i segnali ed i meccanismi che danno inizio alla risposta apoptotica sono ancora in parte da definire. Diversi studi hanno evidenziato che i ceramici, una particolare classe di sfingolipidi, sono implicati nella traduzione dei segnali intracellulari partecipando a diversi processi quali la modulazione del ciclo cellulare, l’induzione del processo di differenziazione e/o del fenomeno apoptotico. I ceramici intracellulari possono venire sintetizzati attraverso varie vie e una di queste implica l’attivazione di particolari enzimi, le sfingomielinasi acida e neutra, che idrolizzano la sfingomielina in fosfocolina e ceramide. Recenti studi in vitro su linee cellulari leucemiche hanno dimostrato che il glutatione (GSH), molecola ad attività antiossidante, inibisce in maniera dose-dipendente l’attivazione della sfingomielinasi neutra (N-SMase). Con il presente studio riportiamo che il pre-trattamento con GSH esogeno inibisce l’attivazione della N-SMase indotta da UVB, riduce la generazione di ceramici intracellulari e blocca parzialmente il processo apoptotico. Cheratinociti umani normali provenienti da interventi di chirurgia plastica sono stati coltivati su fibroblasti 3T3 trattati con mitomicina in terreno di coltura Dulbecco ed Ham F12 addizionato del 10% di siero di vitello fetale. A subconfluenza le cellule sono state pre-trattate con GSH 5mM, per 2h a 37°C e successivamente irradiate con UVB alla dose di 75mJ/ cm2. A tempi diversi le cellule sono state raccolte per valutare l’ attività enzimatica della N-SMase, la produzione di ceramici intracellulari, tramite cromatografia su strato sottile e il clivaggio della proteina poli(ADP-ribosio) polimerasi (PARP), come marker di apoptosi, tramite western-blot. L’esposizione ai raggi UVB induce attivazione della SMase neutra a 15 minuti con picco a 30 minuti dallo stimolo con UVB e idrolisi della sfingomielina con rapida comparsa di ceramici. L’aumento dei ceramici è risultato significativamente evidente già dopo 15 minuti dall’irradiazione UV e si mantiene fino a 24 ore. Il pre-trattamento con GSH inibisce, in modo significativo, l’attivazione in vitro della N-SMase e la produzione di ceramici a tutti i tempi studiati. L’analisi in western-blotting ha evidenziato il clivaggio di PARP a 16 ore dall’esposizione ai raggi UVB, mentre nei campioni protrattati con GSH non si osserva tale clivaggio evidenziando in questo modo un’interferenza con la trasduzione del segnale apoptotico. I nostri dati suggeriscono che il glutatione endogeno sia uno dei fattori che regolano l’attivazione della N-SMase nel processo di apoptosi indotta dai raggi UVB nei cheratinociti umani normali in coltura.

2001 - UVB-radiation induces neutral and acidic sphingomyelinases in cultured normal human keratinocytes. [Abstract in Atti di Convegno]
Euclidi, Emanuela; Magnoni, Cristina; Benassi, Luisa; Bertazzoni, Giorgia; Vaschieri, Cristina; Giannetti, Alberto; Seidenari, Stefania
abstract

The sphingomyelin pathway is an ubiquitous, evolutionary conserved signaling system which transduces extracellular signal into the cell. During the last year increasing evidence has shown that sphingolipids may play a role in intracellular signal transduction. The action of a ligand binding to a surface receptor results in early activation of the enzyme sphingomyelinase (SMases). SMase causes hydrolysis of membrane sphingomyelin (SM) and generation of ceramide. Activation of sphingomyelin cycle has been linked to various extracellular agents such as TNF-α and ionizing radiation. Activation of these targets is followed by three major cellular responses: cell growth arrest, induction of cell differentiation and/or induction of programmed cell death or apoptosis. The aim of the present study is to investigate whether activation of SMases and generation of ceramide could be induced by UVB radiation in normal human keratinocytes. (NHK).NHK were cultivated with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12 medium and, at preconfluency were irradiated with a UVB dose of 75mJ/cm2.At different times after UVB irradiation, cells were harvested for lipid extraction and for in vitro measurement of neutral and acidic SMaese enzymatic activity.Exposure to UVB radiation results in a rapid in vivo sphingomyelin hydrolysis and generation of ceramide as measured by TLC analysis. The ceramide accumulation starts at 15 min after UV exposure and progressively increased up to 24h.In vitro measurement of neutral SMases activity from UVB-treated NHK extracts, using labelled C14 sphingomyelin as substrate, shows that acidic SMase peaks 15 minutes after exposure to UVB, while neutral SMase peaks at 30 minutes. TUNEL shows apoptotic cells in normal human keratinocytes after UVB radiation with a peak at 48 h. These data indicate that UVB can act on cellular membranes inducing sphingomyelin hydrolysis and ceramide production through both neutral and acidic SMases.

2000 - Ceramide 2 (N-acetyl sphingosine) is associated with reduction in Bcl-2 protein levels by Western blotting and with apoptosis in cultured human keratinocytes [Articolo su rivista]
A., Di Nardo; Benassi, Luisa; Magnoni, Cristina; Cossarizza, Andrea; Seidenari, Stefania; Giannetti, Alberto
abstract

Background Ceramides produced by sphingomyelin hydrolysis activate a cycle that is followed by three different major cellular responses: downregulation of cell proliferation, induction of cell differentiation and apoptosis. In the skin, the generation of intracellular ceramide may also provide a link between an extracellular signal and the induction of the apoptosis programme for the elimination of damaged cells. Objectives We investigated the effect of ceramides capable of entering cells on cultured keratinocytes. Methods Human keratinocytes from neonatal skin were cultured in serum-free medium with or without increasing concentrations of ceramide 2 (CER-2; N-acetyl sphingosine) (5, 10, 20 and 40 mu mol L-1). Proliferative effects were studied either by cell counts or by H-3-thymidine incorporation and flow cytometric analysis. Apoptosis was studied by TUNEL staining and Western blot analysis of Bcl-2 protein. Results Cell counts and DNA synthesis were reduced in a dose-dependent manner following CER-2 treatment. TUNEL staining showed CER-2-induced apoptosis at 48, 72 and 96 h. Western blot analysis showed that CER-2 induces downregulation of Bcl-2, at 24-96 h. Conclusions These results demonstrate that CER-2 inhibits cell proliferation and induces apoptosis, possibly via a Bcl-2-dependent mechanism.

2000 - Decrease in toxic potential of tensides when used in association: in vitro study on normal human keratinocytes [Abstract in Atti di Convegno]
Bertazzoni, Giorgia; Benassi, Luisa; Magnoni, Cristina; A., Di Nardo; L., Grenzi; Seidenari, Stefania
abstract

Sodium lauryl sulphate (SLS) is an anionic tenside that is widely utilised as a model for studying acute and cumulative irritation. When used in culture, SLS induces an inhibition of DNA synthesis and a decrease in cellular viability. It has previously been reported that the barrier damage caused by SLS in vivo is lower when SLS is applied to the skin in combination with other tensides. The aim of our study was to evaluate if the cytotoxic effect of SLS is reduced by the association with different tensides also in vitro. Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfluency cells were treated with SLS at a dose corresponding to the IC 50 in combination with cocamidopropylbetaine, Tween 20 and Tween 80 at the minimum toxic dose. Following tenside treatment, the culture medium was changed and after 24 hours the cells were collected for thymidine incorporation, MTT assay and neutral red uptake. The cytotoxic effect on normal human keratinocytes as evaluated by thymidine incorporation, MTT assay and neutral red uptake was significantly decreased by combination with all the tested tensides.

2000 - Interleukin 1α after exposure to SLS is time and dose dependent in normal human keratinocytes in colture [Abstract in Atti di Convegno]
Euclidi, Emanuela; Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Seidenari, Stefania
abstract

Interleukin-1alfa is involved in the biological response to irritants as one of the most important inflammatory mediators. The aim of the present study was to evaluate IL-1alfa production after exposure to sodium lauryl sulphate (SLS) from cultured keratinocytes, representing a model which enables the assessment of the in vitro effects of SLS epidermal cells independently from the skin immune system and barrier alterations. Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfluency, cells were treated with SLS doses ranging from 0.00001% to 0.005%. After one hour exposure the medium was changed. At different time-points the supernatant was collected for ELISA, and cells were harvested for Western blot analysis of pro-IL-1alfa and IL-1 alfa. Extracellular secretion of IL-1alfa from keratinocytes was increased in a dose-dependent manner following SLS treatment. The release of IL-1alfa starts at 15 minutes after exposure to SLS (IC50) reaching a maximum at 3 hours. Western blot analysis showed a down regulation of pro-IL-1alfa at 1 hour. Decrease was evident 1 hour after SLS treatment and was maximal at 12 hours.

2000 - L'esposizione ai raggi UVB induce l'attivazione delle sfingomielinasi neutra e acida in cheratinociti umani normali in coltura. [Abstract in Atti di Convegno]
Euclidi, Emanuela; Magnoni, Cristina; A., Di Nardo; Benassi, Luisa; Bertazzoni, Giorgia; Vaschieri, Cristina; Seidenari, Stefania; Giannetti, Alberto
abstract

Il pathway della sfingomielina, ubiquitario e conservato dal punto di vista evoluzionistico, partecipa alla trasduzione del segnale dall’esterno all’interno della cellula. La sfingomielina (SM) è uno sfingolipide concentrato preferenzialmente nella porzione esterna della membrana citoplasmatica delle cellule di mammifero, ma presente anche nella parte interna della membrana in piccola quota dove sembra essere implicata nei processi di trasduzione del segnale. In questo sistema, lo stimolo di alcuni recettori presenti sulla superficie cellulare attiva una famiglia di enzimi specifici, denominati sfingomielinasi, che idrolizzano la SM generando fosfocolina e ceramide.Due tipi di sfingomielinasi (SMasi) sono coinvolte in questo processo ed agiscono a pH neutro o acido. La SMasi neutra idrolizza la sfingomilelina nell’ambiente extracellulare, mentre la SMasi acida agisce principalmente all’interno dei lisosomi.E’ stato osservato in diverse linee cellulari che uno dei primi eventi del processo apoptotico indotto da TNF-, o da radiazioni ionizzanti, é rappresentato da una rapida attivazione di entrambe le SMasi con conseguente produzione di ceramidi .Inoltre dati recenti dimostrano che il trattamento con analoghi dei ceramidi esogeni é sufficiente per l’ induzione dell’apoptosi nei cheratinociti umani normali.Lo scopo del presente studio è quello di valutare il ruolo delle SMasi neutra e acida e dei ceramidi nel segnale intracellulare generato dai raggi UVB in cheratinociti umani normali.Cheratinociti umani normali provenienti da interventi di chirurgia plastica sono stati coltivati su fibroblasti 3T3 mitomicinati con terreno di coltura Dulbecco ed Ham F12 addizionato del 10% di siero di vitello fetale. A subconfluenza le cellule sono state irradiate con UVB alla dose di 100mJ/ cm2 e raccolte a tempi per il dosaggio dei lipidi intracellulari e per il dosaggio in vitro dell’ attività enzimatica delle sfingomielinasi.Il dosaggio dei lipidi intracellulari tramite TLC ha evidenziato che la esposizione a UVB induce rapida idrolisi della sfingomielina e aumento della quota di ceramidi. L’aumento dei ceramidi é risultato significativamente evidente dopo 15 minuti dall’esposizione con UVB con incremento successivo fino alle 24h. Il dosaggio delle attività enzimatiche è stato effettuato usando come substrato sfingomielina marcata con C14. La attività della SMasi acida mostra un picco a 15 minuti dalla esposizione ai raggi UVB, mentre per la SMase neutra il picco era presente a 30 minuti.Dopo 48h il numero di cellule apoptotiche erano significativamente alte rispetto al controllo.Questi dati, indicano che i raggi ultravioletti possono agire sulla membrana cellulare inducendo idrolisi della SM e conseguente produzione di ceramidi attraverso la attivazione sia della SMasi acida che di quella neutra.

2000 - Sintesi e rilascio di Interleuchina 1 alfa in seguito all'esposizione di sodio lauril solfato in cheratinociti umani normali. [Abstract in Atti di Convegno]
Bertazzoni, Giorgia; Benassi, Luisa; Magnoni, Cristina; A., Di Nardo; Euclidi, Emanuela; Vaschieri, Cristina; Giannetti, Alberto; Seidenari, Stefania
abstract

La risposta cellulare agli irritanti si accompagna alla produzione ed al rilascio di numerosi mediatori infiammatori tra cui l’interleuchina 1alfa(IL-1alfa). Il rilascio di questa citochina può quindi essere studiato come marker di tossicità acuta indotta dai detergenti.Lo scopo di questo lavoro è stato valutare la sintesi e il rilascio di IL-1alfa da parte di cheratinociti umani normali in coltura dopo esposizione a Sodio Lauril Solfato (SLS).I cheratinociti, estratti da cute proveniente da interventi di chirurgia plastica, sono stati messi in coltura con un mezzo definito privo di siero. A subconfluenza, sono stati trattati con concentrazioni di SLS variabili da 0,00001 a 0,01%. Dopo un’ora il mezzo di coltura è stato cambiato e a tempi differenti sono stati raccolti i sovranatanti per l’esecuzione del test ELISA e i lisati cellulari per l’analisi in Western Blot e Northern Blot.Dai risultati ottenuti è emerso che l’esposizione a SLS induce il rilascio di IL-1alfa nel sovranatante in modo dose-dipendente, raggiungendo il valore massimo alla concentrazione di 0,0025%, (corrispondente all’IC50). Il rilascio di IL-1alfa è risultato inoltre aumentato già dopo 15 min dal trattamento con valori massimi alle 24 ore.I livelli intracellulari di IL-1alfa diminuiscono nella prima ora dopo il trattamento, per poi aumentare e raggiungere valori significativamente alti a partire dalle 12 ore.La cinetica della sintesi di mRNA mostra un andamento bifasico, evidenziando un aumento immediato al termine dell’esposizione all’SLS e un secondo picco a 12 ore.

2000 - Thickness and echogenicity of the skin in children as assessed by 20-MHz ultrasound [Articolo su rivista]
Seidenari, Stefania; G., Giusti; Bertoni, Laura; Magnoni, Cristina; Pellacani, Giovanni
abstract

Background: Skin anatomy and physiology undergo modifications throughout the whole lifespan. in children the skin appears with structural characteristics, as evaluated by histology, which differ from those of adults, especially in the first years of life. Objective: The aim of our study was to evaluate skin thickness and echogenicity at different sites in children by 20-MHz ultrasound. Methods: Eight skin sites of 42 children and 30 young adults were studied with a 20-MHz B scanner. Skin thickness and mean echogenicity were evaluated. The echographic images were processed and segmented by a dedicated program: the 0-30 amplitude interval, which marks the hypoechogenic parts of the tissue, and the 201-255 range, highlighting the hyperreflecting areas, have been selected. Results and Conclusion: Whereas skin thickness shows a gradual increase from birth to adulthood, maturation of the skin leads to variations in the intensity of its echogenicity, depending on the different skin areas. Whereas on the face and the trunk it appears lower in adults with respect to children, a gradual increase can be observed on the limbs with growing age. The distribution of skin reflectivity also greatly varies in different phases of life. Copyright (C) 2000 S. Karger AG, Basel.

1999 - Ceramide 2(N-acetyl sphingosine) induces G1 arrest and Rb dephosphorilation in cultured human keratinocytes [Abstract in Rivista]
Di Nardo, A; Magnoni, Cristina; Benassi, Luisa; Bertazzoni, Giorgia; Seidenari, Stefania; Giannetti, Alberto
abstract

In the skin, the generation of intracellular ceramide may provide a link between an extracellular signal and the induction of apoptosis for the elimination of damaged cells. Previous sturdies have shown that cell permeant ceramide are involved in signal transduction, cell cycle regulation and apoptosis in different cell types. The aim of the present study was to investigate the intracellular apoptotic signal induced by permeant ceramides on cultured normal human keratinocytes (NHK). NHK from neonatal skin were cultured in serum free medium with or without ceramide 2 (CER-2, n-acetil-sphingosine) 20 M, 40 M. Cell cycle was investigated by FACS analysis, and western blot analysis of cyclin D1 and Rb dephosphorilation. Apoptosis was studied by TUNEL staining and western blot analysis of BCL-2 protein, p53 and p21. FACS analysis demonstrated a G1 arrest 48 h after ceramide addition, accompanied by down regulation of cyclin D1 and by Kb dephosphorilation. TUNEL staining showed the presence of CER-2-induced apoptosis after 48, 72 and 96 h of culture. Western blot analysis demonstrated that CER-2 induces a down regulation of BCL-2 after 24 h up to 96 h, and an up regulation of p53 after 24 h. No modifications were noted in p21 levels. These results demonstrate that in NHK CER-2 cell permeant ceramide induces cell cycle arrest in G1, which precedes apoptosis.

1999 - Curva dose-risposta e time-course del rilascio di interleuchina 1-alpha su colture di cheratinociti umani normali dopo esposizione a differenti concentrazioni [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; L., Grenzi; Seidenari, Stefania
abstract

La complessa risposta cellulare agli irritanti si accompagna alla produzione e al rilascio di numerosi mediatori, tra cui l'interleuchina 1alfa (IL-1 alfa). Lo scopo del nostro studio è stato quello di valutare la liberazione di IL-1 alfa dai cheratinociti dopo esposizione al Sodio Lauril Solfato (SLS), un detergente anionico, indipendentemente dalla stimolazione del sistema immunitario e dalle alterazioni di barriera che si producono in vivo per esposizione della cute a questo tensioattivo. Abbiamo pertanto utilizzato cheratinociti umani normali in coltura monostrato che sono stati trattati con SLS a concentrazioni variabili tra 0,00001 e 0,005% diluito in mezzo definito privo di siero (KBM). Dopo un'ora di esposizione, il mezzo è stato cambiato e dopo 24 h il sovranatante è stato raccolto per l'esecuzione di test ELISA (Enzyme Linked Immunoassorbent Assay). Le cellule sono state inoltre lisate per analisi in Western blot della pro-IL-1 alfa (30kD) e della IL-1 alfa (17kD). Per quanto riguarda la curva dose-risposta, l'ELISA ha mostrato che la secrezione extracellulare di IL-1 alfa aumenta con l'incremento della dose SLS. Per quanto riguarda il time-course invece, il rilascio di IL-1alfa nel sovranatante è aumentato da minuti a 3 ore dopo lo stimolo. L'analisi Westerb blot delle proteine citoplasmatiche ha mostrato parallelamente un decremento della IL-1alfa accompagnato da un andamento costante della forma a 17 kD.

1999 - Decrease in toxic potential of tensides when used in association: in vitro study on normal human keratinocytes [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; L., Grenzi; Seidenari, Stefania
abstract

Sodium lauryl sulfate is an anionic tenside that is widely utilised as a model for studying acute and cumulative irritation. When used in culture, SLS induces an inhibition of DNA synthesis and a decrease in cellular viability. It has been previously reported that the barrier damage caused by SLS in vivo is lower when SLS is applied to the skin in combination with other tensides.The aim of our study was to evaluate if the cytotoxic effect of SLS is reduced by the association with different tensides also in vitro.Normal human keratinocytes from plastic surgery were grown in serum-free medium. At subconfluency, cells were treated with SLS at a dose corresponding to the DL50 in combination with Tween 20, Tween 80, Tegobetaine F50 at the minimum toxic dose.Following tenside treatment, the culture medium was changed, and after 24 hours the cells were collected for thymidine incorporation, MTT assay and NR uptake.The cytotoxic effect on normal human keratinocytes as evaluated by thymidine incorporation, MTT assay and NR uptake was significantly decreased by the combination with all the tested tensides.

1999 - Dose response curves of interleukin 1 alpha release by cultured normal human keratinocyte after variable exposure to different concentrations of SLS [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Seidenari, Stefania
abstract

One of the primary events or response of human keratinocyte to irritant is the release of a variety of mediators. Among these, interleukin 1 alpha (IL-1alfa) has been shown to be increased following different stimuli induced by inflammatory agents or microbial endotoxins. Furthermore, it has been shown that IL-1 alfa secretion is triggered by exposure to tensides.To study the direct effects of irritants on keratinocytes without the involvement of inflammatory cells and the secondary effects of barrier disruption, we used an in vitro culture model.In this preliminary study, we investigated the in vitro release of IL-1 alfa produced by cultured normal human keratinocytes (NHK) treated with Sodium Lauryl Sulfate (SLS), an anionic detergent, at concentrations ranging from 0,00001 to 0,005. diluited in a protein free medium (KBM).After 1 h exposure, cells were washed and the medium was changed. After 24 h the supernatant was collected for Enzyme Linked Immunoassorbent Assay (ELISA).Results presented here are the mean of three independent experiments.Our results show that extracellular secretion of IL-1 alfa has an increasing trend with increasing SLS doses and exposure times.

1999 - Effects of tensides on normal human keratinocytes: A decrease in the toxic potential when used in association [Abstract in Rivista]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; Di Nardo, A; Grenzi, L; Seidenari, Stefania
abstract

Sodium lauryl sulfate is an anionic tenside that is widely utilised as a model for studying acute and cumulative irritation. When used in culture, SLS induce an inhibition of DNA synthesis and a decrease in cellular viability. It has been previously reported that the barrier damage caused by SLS in vivo is lower when SLS is applied to the skin in combination with other tensides.The aim of our study was to evaluate if the cytotoxic effects of SLS is reduced by the association with different tensides olso in vitro.Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfluency cells were treated with SLS at a dose corresponding to the DL50 in combination with Tween 20. Tween 80, Tegobetaine F50 at the minimum toxic dose.Following tenside treatment, the culture medium was changed and after 24 h the cells were collected for thymidine incorporation, MTT assay and NR uptake.The cytotoxic effect on normal human keratinocytes as evaluated by thymidine incorporation, MTT assay and NR uptake was significantly decreased by the combination with all the tested tensides.

1999 - Effects of tensides on normal human keratinocytes: a decrease in toxic potential when used in association [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; L., Grenzi; Seidenari, Stefania
abstract

Sodium lauryl sulfate is an anionic tenside that is widely utilised as a model for studying acute and cumulative irritation. When used in culture, SLS induce an inhibition of DNA synthesis and a decrease in cellular viability. It has previously been reported that the barrier damage caused by SLS in vivo is lower when SLS is applied to the skin in combination with other tensides.The aim of our study was to evaluate wheter the cytotoxic effects of SLS is reduced by the association with various tensides in vitro.Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfluency cells were treated with SLS at a dose corresponding to effective dose 50%, in combination with Tween 20, Tween 80, Tegobetaine F50 at the minimum toxic dose.Following tenside treatment, the culture medium was changed and after 24 h the cells were collected for thymidine incorporation, MTT assay and neutral red uptake.The cytotoxic effect on normal human keratinocytes as evaluated by thymidine incorporation, MTT assay and NR uptake was significantly decreased by the combination with all the tested tensides.

1999 - G1 arrest and Rb dephosphorilation is induced by ceramide 2(N-acetyl sphingosine) in cultured human keratinocytes [Abstract in Atti di Convegno]
A., Di Nardo; Magnoni, Cristina; Benassi, Luisa; Bertazzoni, Giorgia; Seidenari, Stefania; Giannetti, Alberto
abstract

In the skin, the generation of intracellular ceramide may provide a link between an extracellular signal and the induction of apoptosis for the elimination of damaged cells.Previous studies have shown that cell permeant ceramides are involved in signal transduction, cell cycle regulation and apoptosis in different cell types. The aim of the present study was to investigate the intracellular apoptotic signal induced by permeant ceramides on cultured normal human keratinocytes (NHK). NHK from neonatal skin were cultured in serum-free medium with or without Ceramide 2 (CER-2, n-acelyl-sphingosine) 20 M, 40 M. Cell cycle was investigated by FACS analysis, and western blot analysis of cyclin D1 and Rb dephosphorilation. Apoptosis was studied by TUNEL staining and western blot analysis of BCL-2 protein, p53 and p21. FACS analysis demonstrated a G1 arrest 48 h after ceramide addition, accompanied by down-regulation of cyclin D1 and by Rb dephosphorilation. TUNEL staining showed the presence of CER-2-induced apoptosis after 48, 72 and 96 h of culture. Western blot analysis demonstrated that CER-2 induces a down-regulation of BCL-2 after 24 h up to 36 h, and an up-regulation of p53 after 24 h. No modifications were noted in p21 levels. These results demonstrate that in NHK CER-2 cell permeant ceramide induces cell cycle arrest in G1, which precedes apoptosis.

1999 - L'utilizzo di tensioattivi in combinazione diminuisce l'effetto citotossico del sodio lauril solfato su cheratinociti umani normali in coltura [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Euclidi, Emanuela; Seidenari, Stefania
abstract

I1 sodio lauril solfato (SLS) un tensioattivo anionico utilizzato come irritante di riferimento sia in vivo che in vitro. Quando utilizzato in colture di cheratinociti umani normali, il SLS induce arresto della proliferazione e diminuzione della vitalità cellulare con danno di membrana. Sulla base di precedenti studi in vivo che dimostrano una diminuzione del danno di barriera con l'uso di combinazioni di tensioattivi rispetto all'utilizzo del solo SLS, abbiamo voluto verificare se anche in vitro fosse osservabile lo stesso effetto. Cheratinociti umani normali sono stati coltivati in mezzo definito privo di siero. Alla subconfluenza le cellule sono state esposte contemporaneamente alla DL50 di SLS (0,0025%) in corrispondenza con concentrazioni di Tegobetaina F50, Tween 20, Tween 80 a bassa tossicità. Dopo un'ora di esposizione ai tensioattivi, il mezzo di coltura è stato cambiato e dopo 24 ore le cellule sono state raccolte per i seguenti saggi: conta cellulare, incorporazione di timidina triziata, rosso neutro e MTT. In tutti i casi le miscele di tensioattivi hanno dato luogo a minori effetti tossici rispetto al trattamento con solo SLS, con minore inibizione della proliferazione e più scarsa compromissione della vitalità cellulare.

1999 - Sodium Lauryl Sulfate causes IL-1α release from normal human keratinocytes: dose response and time course curves [Abstract in Rivista]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Euclidi, Emanuela; Seidenari, Stefania
abstract

Interleukin 1alfa (IL-1 alfa) is involved in the biological response to irritant as one of the most important inflammatory mediator. The aim of the present study was to evaluate IL-1 alfa production after exposure to Sodium Lauryl Sulfate (SLS) from cultured human keratinocytes, representing a model which enables to asses the in vitro effects of SLS on epidermal cells independently from the skin immune system and barrier alteration.Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfuency cells were treated with SLS doses ranging from 0.00001 to 0.005%. After one hour exposure the medium was changed. At different time points the supernatant was collected for ELISA, and cells were harvested for Western blot analysis of pro-IL-1 alfa and IL-1 alfa. Extracellular secretion of IL-1alfa from keratinocytes was increased in a dose dependent manner following SLS treatment. The release of IL-1alfa starts at 30 min after exposure to SLS (DL50) reaching a maximum at 3 h.Western blot analysis showed a down regulation of pro-IL-1alfa levels at 1 h, while IL-1 alfa levels remained unchanged.

1999 - Sodium lauryl sulfate causes interleukin-1α release from normal human keratinocytes: dose-response and time-course curves [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Euclidi, Emanuela; Seidenari, Stefania
abstract

Interleukin 1alfa (IL-1 alfa) is involved in the biological response to irritants as one of the most important inflammatory mediators. The aim of the present study was to evaluate IL-1 alfa production after exposure of cultured human keratinocytes to Sodium Lauryl Sulfate (SLS), representing a model which enables assessment of the in vitro effects of SLS on epidermal cells independently from the skin immune system and barrier alteration.Normal human keratinocytes from plastic surgery were grown in serum free medium. At subconfuency cells were treated with SLS doses ranging from 0.00001 to 0.005% and after exposure for one hour, the medium was changed. At different time points the supernatant was collected for ELISA, and cells were harvested for Western blot analysis of pro-IL-1 alfa and IL-1 alfa. Extracellular secretion of IL-1 alfa from keratinocytes was increased in a dose dependent manner following SLS treatment. The release of IL-1alfa starts at 30 min after exposure (effective dose 50%) reaching a maximum at 3 h.Western blot analysis showed a down regulation of pro-IL-1alfa levels, while IL-1 alfa levels remained unchanged.

1999 - Sodium lauryl sulphate causes IL-1α release as a marker of irritant response in normal human keratinocytes: dose response and time course curves. [Abstract in Atti di Convegno]
Benassi, Luisa; Bertazzoni, Giorgia; Magnoni, Cristina; A., Di Nardo; Euclidi, Emanuela; Seidenari, Stefania
abstract

Interleukin 1 alfa is involved in the biological response to irritants as one of the most important inflammatory mediators. The aim of the present study was to evaluate IL-1 alfa production after exposure to Sodium Lauryl Sulfate (SLS) from cultured human keratinocytes, representing a model which makes it possible to assess the in vitro effects of SLS on epidermal cells independently from the skin immune system and barrier alterations.Normal human keratinocytes from plastic surgery were grown in serum-free medium. At subconfluency, cells were treated with SLS doses ranging from 0.00001 to 0.005%, and after a one-hour exposure the medium was changed.At different time points, the supernatant was collected for ELISA, and cells were harvested for Western blot analysis of pro-IL -1 alfa and IL-1 alfa .Extracellular secretion of IL-1 alfa from keratinocytes was increased in a dose-dependent manner following SLS treatment. The release of IL-1alfa starts at 30 minutes after exposure to SLS (DL 50), reaching a maximum at 3 hours.Western blot analysis showed a down-regulation of pro IL-1 alfa levels at 1 hour, while IL-1 alfa levels remained unchanged.

1999 - UVB radiation induces neutral and acidic sphingomyelinases in normal human keratinocytes: trigger for ceramide pathway. [Abstract in Atti di Convegno]
Magnoni, Cristina; A., Di Nardo; Benassi, Luisa; Euclidi, Emanuela; Seidenari, Stefania; Giannetti, Alberto
abstract

The sphingomyelin pathway is an ubiquitous, evolutionary-conserved signaling system which transduces extracellular signal into the cell. Cleavage of sphingomyelinase (SMases), evoked by different stimuli, causes the release of ceramide, which can act as a second messenger for mediating apoptosis. Sphingomyelin can be hydrolysed by two types of SMasewith neutral or acidic pH optimum, respectively. We have shown previously that treatment with exogenous ceramides analogues was sufficient for induction of apoptosis in normal human keratinocytes. The aim of the present study was to investigate the role of neutral and acidic SMases and ceramide in the intracellular signalling generated after UVB exposure of normal human keratinocytes(NHK). NHK were cultivated with mitomycin-treated 3T3 cells in Dulbecco’s modified Eagle’s medium/Ham’s F12 medium and, at preconfluency were irradiated with a UVB dose of 100mJ/cm2.At different times after UVB irradiation, cells were harvested for lipid extraction and for in vitro measurement of neutral and acidic SMaese enzymatic activity.Exposure to UVB radiation results in a rapid in vivo sphingomyelin hydrolysis and generation of ceramide as measured by TLC analysis. The ceramide accumulation peaked at 15 min after UVB irradiation.In vitro measurement of neutral SMases activity from UVB-treated NHK extracts, using labelled sphingomyelin as substrate, showed an induction of both neutral and acidic SMase with slightly different kinetics. These data indicate that UVB can act on cellular membranes, inducing sphingomyelin hydrolysis and ceramide production through both neutral and acidic SMases.

1998 - C2-ceramide triggers a BCL-2 mediated apoptosis in normal human keratinocytes. [Abstract in Rivista]
Di Nardo, A; Benassi, Luisa; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1998 - Thymidine dimers induce a prolonged P53-dependent cell cycle arrest but not apoptosis in cultured human keratinocytes [Abstract in Atti di Convegno]
Magnoni, Cristina; Benassi, Luisa; A., Di Nardo; Bertazzoni, Giorgia; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - Acetyl-ceramide induced apoptosis in cultured human keratinocytes. [Abstract in Rivista]
Dinardo, A; Benassi, Luisa; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

.

1997 - Autocrine nerve growth factor protects human keratinocytes from apoptosis through its high affinity receptor (TRK): A role for BCL-2 [Articolo su rivista]
Pincelli, Carlo; A. R., Haake; Benassi, Luisa; E., Grassilli; Magnoni, Cristina; D., Ottani; R., Polakowska; C., Franceschi; Giannetti, Alberto
abstract

Normal human keratinocytes synthesize and release nerve growth factor (NGF) and express both the low-and the high-affinity NGF receptor, Because NGF has been shown to rescue certain cell types from programmed cell death, we investigated the role of endogenous NGF in preventing keratinocyte apoptosis, We report here that apoptosis is induced in normal human keratinocytes in culture by blocking endogenous NGF signaling with either anti-NGF neutralizing antibody or K252, a specific inhibitor of the tyrosine kinase high-affinity NGF receptor. Apoptosis was assessed by DNA laddering, electron microscopy, and in situ nick end labeling technique, In anti-NGF-treated keratinocytes, the apoptotic process starts at 96 h, and is maximal at 120 h. After K252 treatment, apoptosis starts at 48 h and peaks at 120 h, Because the product of the bcl-2 proto-oncogene protects many cell types from apoptosis, we measured the levels of this protein in apoptotic keratinocytes, We found that both K252 and anti-NGF antibody strikingly downregulate bcl-2 expression, starting at 72 h. Furthermore, HaCat keratinocytes stably transfected with a plasmid containing bcl-2 cDNA fail to undergo apoptosis when treated with K252. These findings show that autocrine NGF acts as a survival factor for human keratinocytes in vitro through its high-affinity NGF receptor, possibly by maintaining constant levels of Bcl-2.

1997 - Enhancement of DNA repair in human skin cells by thymidine dinucleotides: evidence for a p53-mediated mammalian SOS response. [Articolo su rivista]
M. S., Eller; T., Maeda; Magnoni, Cristina; D., Atwal; B. A., Gilchrest
abstract

Thymidine dinucleotide (pTpT) stimulates melanogenesis in mammalian pigment cells and intact skin, mimicking the effects of UV irradiation and UV-mimetic DNA damage. Here it is shown that, in addition to tanning, pTpT induces a second photoprotective response, enhanced repair of UV-induced DNA damage. This enhanced repair results in a 2-fold increase in expression of a UV-damaged chloramphenicol acetyltransferase expression vector transfected into pTpT-treated skin fibroblasts and keratinocytes, compared with diluent-treated cells. Direct measurement of thymine dimers and (6-4) photoproducts by immunoassay demonstrates faster repair of both of these UV-induced photoproducts in pTpT-treated fibroblasts. This enhanced repair capacity also improves cell survival and colony-forming ability after irradiation. These effects of pTpT are accomplished, at least in part, by the up-regulation of a set of genes involved in DNA repair (ERCC3 and GADD45) and cell cycle inhibition (SDI1). At least two of these genes (GADD45 and SDI1) are known to be transcriptionally regulated by the p53 tumor suppressor protein. Here we show that pTpT activates p53, leading to nuclear accumulation of this protein, and also increases the specific binding of this transcription factor to its DNA consensus sequence.

1997 - I dinucleotidi della timidina aumentano la capacità riparativa del DNA e non inducono apoptosi nei cheratinociti umani in coltura. [Abstract in Atti di Convegno]
Magnoni, Cristina; M. S., Eller; Benassi, Luisa; A., Di Nardo; Giannetti, Alberto; B. A., Gilchrest
abstract

.

1997 - Thymidine dimers induce cell cycle arrest, but not apoptosis in cultured human keratinocytes. [Abstract in Rivista]
Magnoni, Cristina; Benassi, Luisa; Dinardo, A; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1997 - Ultraviolet radiations induce c-src activation in cultured human keratinocytes. [Abstract in Rivista]
Chiodino, C; Ottani, D; Magnoni, Cristina; Giannetti, Alberto; Pincelli, Carlo
abstract

.

1996 - Autocrine nerve growth factor protects human keratinocytes from apoptosis through its high-affinity receptor (trk): A role for BCL-2 [Abstract in Rivista]
Pincelli, Carlo; Magnoni, Cristina; Grassilli, E; Benassi, Luisa; Ottani, D; Franceschi, Claudio; Giannetti, Alberto
abstract

.

1996 - I dimeri della timidina aumentano il riparo del DNA in cheratinociti e fibroblasti umani normali. [Abstract in Atti di Convegno]
Magnoni, Cristina; M. S., Eller; D., Atwal; C., Chiodino; Giannetti, Alberto; B. A., Gilchrest
abstract

.

1996 - Il nerve growth factor endogeno protegge i cheratinociti umani dall'apoptosi attraverso il suo recettore ad alta affinità (trk):un ruolo per la bcl-2. [Abstract in Atti di Convegno]
D., Ottani; Magnoni, Cristina; E., Grassilli; Benassi, Luisa; Franceschi, Claudio; Giannetti, Alberto; Pincelli, Carlo
abstract

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1996 - Thymidine dinucleotides enhance DNA repair in normal human skin cancer [Abstract in Rivista]
Magnoni, Cristina; M. S., Eller; D., Atwal; B. A., Gilchrest
abstract

.

1995 - Expression and function of nerve growth factor (NGF) and NGF receptor on keratinocytes. [Abstract in Atti di Convegno]
Pincelli, Carlo; Magnoni, Cristina; M., Yaar; B. A., Gilchrest; Giannetti, Alberto
abstract

.

1995 - K252a, a high affinity NGF-receptor (trk) inhibitor, induces apoptosis in cultured human keratinocytes. [Abstract in Atti di Convegno]
Magnoni, Cristina; E., Grassilli; Benassi, Luisa; D., Ottani; Franceschi, Claudio; Pincelli, Carlo; Giannetti, Alberto
abstract

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1995 - K252a, A High-Affinity NGF-Receptor (Trk) Inhibitor, Induces Apoptosis In Human Keratinocytes [Abstract in Rivista]
Pincelli, Carlo; Magnoni, Cristina; E., Grassilli; Benassi, Luisa; D., Ottani; C., Franceschi; Giannetti, Alberto
abstract

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1995 - Nerve Growth-Factor Is Increased In Psoriatic Skin [Articolo su rivista]
F., Fantini; Magnoni, Cristina; L., Bracci Laudiero; Pincelli, Carlo
abstract

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1995 - Nerve Growth-Factor Receptor And Neurochemical Markers In Human Oral-Mucosa - An Immunohistochemical Study [Articolo su rivista]
F., Fantini; Giannetti, Alberto; Benassi, Luisa; V., Cattaneo; Magnoni, Cristina; Pincelli, Carlo
abstract

Background: The innervation of the oral mucosa has so far been studied mainly by histochemical and ultrastructural techniques. Only few studies have investigated the presence of neural proteins and neurotransmitters in human gingival mucosa. Objective: The purpose of the present study was to evaluate the presence and distribution of neural structural and transmitter proteins in different areas of normal human oral mucosa. Method: Indirect immunofluorescence was employed on specimens taken from different mucosal regions (gingiva, lips, gums, palate). Both structural (low-affinity nerve growth factor receptor, NGFr; protein gene product 9.5, PGP 9.5) and neuropeptide markers (substance P; calcitonin gene-related peptide; vasoactive intestinal peptide, neuropeptide Y) were used. Results: NGFr and PGP 9.5 intensely labelled both nerve fibres and selected epithelial cells, while neuropeptide immunoreactivity was scarcely expressed and exclusively localized in nerve fibres, Conclusion: Similarity in the distribution pattern and neurochemistry between oral and cutaneous innervation is apparent. Expression of NGFr could be relevant to the trophism of both the oral innervation and epithelium.

1995 - Neurogenic Inflammation And The Skin Neural Modulation Of Cutaneous Inflammatory Reactions [Articolo su rivista]
F., Fantini; Magnoni, Cristina; Pincelli, Carlo; Giannetti, Alberto
abstract

This review presents current concepts in neurogenic inflammation, with particular regard to the skin. Although the participation of the peripheral nervous system in the development of inflammatory reactions has been known for a long time, recent advances in the understanding of the distribution, functions and targets of neural mediators has greatly enhanced interest and research on this topic. A role for peripheral nerve terminals not only in the vascular responses, but also in the modulation of inflammatory cell activities, has been proposed. Neuropeptides released from nerve endings could intervene in the various phases of the inflammatory process, contributing to the complex network of inflammatory mediators which locally regulate cell function, trafficking and trophism. An increasing body of evidence also supports an important role for these fibers in the pathogenesis of both spontaneous and experimentally-induced cutaneous inflammatory lesions. Indeed, recent findings indicate that the different neural transmitters take part in the mechanisms of cutaneous inflammatory reactions with specific, differentiated and often protective effects. Therefore, we would suggest that this involvement could be more properly described as neurogenic modulation of inflammatory reactions.

1995 - UNILATERAL LOCALIZED HYPERHIDROSIS WITH IMMUNOHISTOCHEMICAL EVIDENCE OF VIPERGIC DYSREGULATION [Articolo su rivista]
Fantini, F; Benassi, Luisa; Magnoni, Cristina; Girolomoni, G.
abstract

We report the case of a woman affected by idiopathic, circumscribed hyperhidrosis of the right side of the thorax. No associated systemic abnormalities, or histologic evidence of sweat gland alterations present. Immunohistochemical investigation of skin specimens using neuronal general (protein gene product 9.5, neuron-specific enolase) and neuropeptide markers (substance P, calcitonin gene-related peptide, neuropeptide tyrosine and vasoactive intestinal peptide - VIP) showed marked reduction of periglandular VIP-immunoreactive fibers at the affected side, as compared to contralateral normal skin. We suggest that a localized functional hyperactivity of autonomic VIP-containing fibers may be implanted in the pathogenesis of idiopathic, circumscribed hyperhidrosis.

1994 - Il nerve growth factor è aumentato nella cute psoriasica. [Abstract in Atti di Convegno]
Pincelli, Carlo; F., Fantini; Magnoni, Cristina; E., Distratis; Benassi, Luisa; L., Aloe; Giannetti, Alberto
abstract

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1994 - Il nerve growth factor, ma non la neurotrofina-3 stimola la proliferazione dei cheratinociti umani in coltura. [Abstract in Atti di Convegno]
Magnoni, Cristina; Benassi, Luisa; F., Fantini; Sevignani, C.; Giannetti, Alberto; Pincelli, Carlo
abstract

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1994 - Psoriasis And The Nervous-System [Articolo su rivista]
Pincelli, Carlo; F., Fantini; Magnoni, Cristina; Giannetti, Alberto
abstract

Both clinical and experimental evidence is accumulating on the role of the nervous system in the pathogenesis of psoriasis. Sporadic reports as well as extensive studies indicate that emotional stress can act as an exacerbating event in psoriasis. Moreover, that neurogenic mechanisms are operating in psoriasis is suggested by clinical, pharmacologic and experimental data. We have focused our investigations on the role of vasoactive intestinal peptide (VIP) and substance P (SP) in psoratic lesions using a variety of experimental approaches: 1) receptor autoradiography; 2) immunohistochemistry; 3) radio-imunoassay; 4) human keratinocytes cultures. Our results indicate that an imbalance of VLP and SP exists in psoriatic lesions. and that these neuropeptides exert different and specific effects on human keratinocytes. At present, however, the finding of psoriasis being exacerbated by psychological factors cannot be satisfactorily explained merely by alterations of neuropeptides in the skin.

1992 - Neuropeptidi e cute: il peptide vasoattivo intestinale nelle dermatosi infiammatorie. [Articolo su rivista]
Pincelli, Carlo; Magnoni, Cristina; F., Fantini; Giannetti, Alberto
abstract

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1992 - Notalgia parestesica. Descrizione di un caso con studio immunoistochimico dell'innervazione cutanea. [Articolo su rivista]
G., Girolomoni; R., Magni; Magnoni, Cristina; F., Fantini
abstract

A case of notalgia paresthesica in a 62-year-old woman is reported. The patient complained of a intense itching, and occasionally burning, brown macule of six years duration on the back. Histologic examination showed increased melanin deposition in the basal layer of epidermis, numerous dermal melanophages, and scattered necrotic keratinocytes in both the epidermis and stratum corneum.Immunohistochemistry on frozen section revealed normal number and distribution of nerve fibers reactive for the general neural markers PGP 9.5 and neuron- specific enolase, and for the neuropeptides substance P-, vasoactive intestinal peptide-,and neuropeptide Y. Topical treatment with capsaicin improved pruritus and paresthetic sensations.