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Chiara PAOLINELLI DEVINCENZI
Personale tecnico amministrativo
Dipartimento di Scienze della Vita sede ex-Scienze Biomediche
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Pubblicazioni
2013
- Ectopic calcification in β-thalassemia patients is associated with increased oxidative stress and lower MGP carboxylation
[Articolo su rivista]
Boraldi, Federica; Garcia Fernandez, M.; PAOLINELLI DEVINCENZI, Chiara; Annovi, Giulia; Schurgers, L.; Vermeer, C.; Cianciulli, P.; Ronchetti, I.; Quaglino, Daniela
abstract
A number of beta-thalassemia (β-thal) patients in the course of the disease exhibit ectopic calcification affecting skin, eyes and the cardiovascular system. Clinical and histopathological features have been described similar to those in pseudoxanthoma elasticum (PXE), although different genes are affected in the two diseases. Cultured dermal fibroblasts from β-thal patients with and without PXE-like clinical manifestations have been compared for parameters of redox balance and for the expression of proteins, which have been already associated with the pathologic mineralisation of soft connective tissues. Even though oxidative stress is a well-known condition of β-thal patients, our results indicate that the occurrence of mineralized elastin is associated with a more pronounced redox disequilibrium, as demonstrated by the intracellular increase of anion superoxide and of oxidized proteins and lipids. Moreover, fibroblasts from β-thal PXE-like patients are characterized by decreased availability of carboxylated matrix Gla protein (MGP), as well as by altered expression of proteins involved in the vitamin K-dependent carboxylation process. Results demonstrate that elastic fibre calcification is promoted when redox balance threshold levels are exceeded and the vitamin K-dependent carboxylation process is affected decreasing the activity of MGP, a well-known inhibitor of ectopic calcification. Furthermore, independently from the primary gene defect, these pathways are similarly involved in fibroblasts from PXE and from β-thal PXE-like patients as well as in other diseases leading to ectopic calcification, thus suggesting that can be used as markers of pathologic mineralisation.
2009
- Fibroblast protein profile analysis highlights the role of oxidative stress and vitamin K recycling in the pathogenesis of pseudoxanthoma elasticum
[Articolo su rivista]
Boraldi, Federica; Annovi, Giulia; Guerra, Deanna; Paolinelli Devincenzi, Chiara; Maria Inmaculada Garcia, Fernandez; Panico, Fulvio; De Santis, Giorgio; Tiozzo, Roberta; Ronchetti, Ivonne; Quaglino, Daniela
abstract
Pseudoxanthoma elasticum (PXE) is a genetic disorder associated to mutations in the ABCC6 gene, however the pathogenetic mechanisms leading to elastic fibre calcification and to clinical manifestations are still unknown. Dermal fibroblasts, directly involved in the production of the extracellular milieu, have been isolated from healthy subjects and from patients affected by PXE, cultured in vitro and characterized for their ability to produce reactive oxygen species, for structural and functional properties of their cell membranes, for changes in their protein profile. Data demonstrate that oxidative stress has profound and endurable consequences on PXE fibroblast phenotype being responsible for: reduced levels of global DNA methylation, increased amount of carbonylated proteins and of lipid peroxidation products, altered structural properties of cell membranes, modified protein expression. In conclusion, these data shed new light on pathogenetic pathways in PXE, by identifying a network of proteins affecting elastic fibre calcification through inefficient vitamin K recycling, and highlighting the role of several differentially expressed proteins that could be regarded as targets for validating the efficacy of future therapeutic strategies aiming to delay and/or revert the pathologic phenotype of PXE fibroblasts. Moreover, these data open new perspectives for investigating PXE-like phenotypes in the absence of ABCC6 mutations.
2008
- Comparison of Fibroblasts from Patients Affected by Pseudoxanthoma elasticum (PXE) or by β-Thalassemia with (β-thal-PXE+) and without PXE-Like Clinical Manifestations (β-thal-PXE-)
[Abstract in Rivista]
Boraldi, Federica; PAOLINELLI DEVINCENZI, Chiara; M. I., GARCIA FERNANDEZ; Annovi, Giulia; Quaglino, Daniela; Tiozzo, Roberta; P., Cianciulli; Ronchetti, Ivonne
abstract
The role of altered redox balance in the pathogenesis of PXE-like clinical manifestations in patients affected by beta-thalassemia is discussed.
2008
- Parameters of oxidative stress are present in the circulation of PXE patients
[Articolo su rivista]
M. I., Garcia Fernandez; Gheduzzi, Dealba; Boraldi, Federica; PAOLINELLI DEVINCENZI, Chiara; P., Sanchez; P., Valdivielso; M. J., Morilla; Quaglino, Daniela; Guerra, Deanna; Casolari, Sara; L., Bercovitch; Ronchetti, Ivonne
abstract
Pseudoxanthoma elasticum (PXE) is an inherited disorder characterized by calcification of elastic fibres leading to dermatological and vascular alterations associated to premature aged features and to life threatening clinical manifestations. The severity of the disease is independent from the type of mutation in the ABCC6 gene, and it has been suggested that local and/or systemic factors may contribute to the occurrence of clinical phenotype. The redox balance in the circulation of 27 PXE patients and of 50 healthy subjects of comparable age was evaluated by measuring the advanced oxidation protein products (AOPP), the lipid peroxidation derivatives (LOOH), the circulating total antioxidant status (TAS), the thiol content and the extracellular superoxide dismutase activity (EC-SOD). Patients were diagnosed by clinical, ultrastructural and molecular findings. Compared to control subjects, PXE patients exhibited significantly lower antioxidant potential, namely circulating TAS and free thiol groups, and higher levels of parameters of oxidative damage, as LOOH and of AOPP, and of circulating EC-SOD activity. Interestingly, the ratio between oxidant and antioxidant parameters was significantly altered in PXE patients and related to various score indices. This study demonstrates, for the first time, that several parameters of oxidative stress are modified in the blood of PXE patients and that the redox balance is significantly altered compared to control subjects of comparable age. Therefore, in PXE patients the circulating impaired redox balance may contribute to the occurrence of several clinical manifestations in PXE patients, and/or to the severity of disease, thus opening new perspectives for their management.
2008
- The effect of serum withdrawal on the protein profile of quiescent human dermal fibroblasts in primary cell culture
[Articolo su rivista]
Boraldi, Federica; Annovi, Giulia; PAOLINELLI DEVINCENZI, Chiara; Tiozzo, Roberta; Quaglino, Daniela
abstract
The effect of serum deprivation on proliferating cells is well known, in contrast its role on primary cell cultures, at confluence, has not been deeply investigated. Therefore, in order to explore the response of quiescent cells to serum deprivation, ubiquitous mesenchymal cells, as normal human dermal fibroblasts, were grown, for 48 h after confluence, in the presence or absence of 10% FBS. Fibroblast behaviour (i.e. cell morphology, cell viability, ROS production and elastin synthesis) was evaluated morphologically and biochemically. Moreover, the protein profile was investigated by 2-DE and differentially expressed proteins were identified by MS. Serum withdrawal caused cell shrinkage but did not significantly modify the total cell number. ROS production, as evaluated by the dihydroethidium (DH2) probe, was increased after serum deprivation, whereas elastin synthesis, measured by a colorimetric method, was markedly reduced in the absence of serum. By proteome analysis, 41 proteins appeared to significantly change their expression, the great majority of protein changes were related to the cytoskeleton, the stress response and the glycolytic pathway. Data indicate that human dermal fibroblasts in primary cell culture can adapt themselves to environmental changes, without significantly altering cell viability, at least after a few days of treatment, even though serum withdrawal represents a stress condition capable to increase ROS production, to influence cell metabolism and to interfere with cell behaviour, favouring the expression of several age-related features.
2007
- Antioxidant status in the circulation of patients with Pseudoxanthoma elasticum (PXE).
[Abstract in Rivista]
Boraldi, Federica; M. I., GARCIA FERNANDEZ; Gheduzzi, Dealba; PAOLINELLI DEVINCENZI, Chiara; P., Sanchez; P., Valdivielso; M. J., Morilla; Guerra, Deanna; Casolari, Sara; Quaglino, Daniela; Ronchetti, Ivonne
abstract
Several markers of redox balance have been measured in the plasma of PXE patients. Data demonstrate the presence of an altered redox balance not only at a local level (connective tissue fibroblasts), but also in the circulation. A correlation with clinical manifestations is provided.
2007
- Fibroblasts from patients with beta-Thalassemia and Pseudoxanthoma elasticum-like clinical manifestations.
[Abstract in Rivista]
PAOLINELLI DEVINCENZI, Chiara; Boraldi, Federica; M. I., GARCIA FERNANDEZ; Annovi, Giulia; Quaglino, Daniela; Tiozzo, Roberta; Ceccarelli, Daniela; P., Cianciulli; P. L., Forni; Ronchetti, Ivonne
abstract
In vitro studies on the phenotype of dermal fibroblasts from patients with beta-thalassemia and PXE-like clinical manifestations are described and the the role of altered redox balance is discussed
2007
- Matrix Gla Protein involved in elastic fiber calcification in the dermis of pseudoxanthoma elasticum patients.
[Articolo su rivista]
Gheduzzi, Dealba; Boraldi, Federica; Annovi, Giulia; Paolinelli Devincenzi, Chiara; L., Schurgers; C., Vermeer; Quaglino, Daniela; Ronchetti, Ivonne
abstract
Mature MGP (Matrix gamma-carboxyglutamic acid protein) is known to inhibit soft connective tissues calcification. We investigated its possible involvement in pseudoxanthoma elasticum (PXE), a genetic disorder whose clinical manifestations are due to mineralization of elastic fibers. PXE patients have lower serum concentration of total MGP compared to controls (P<0.001). Antibodies specific for the noncarboxylated (Glu-MGP) and for the gamma-carboxylated (Gla-MGP) forms of MGP were assayed on ultrathin sections of dermis from controls and PXE patients. Normal elastic fibers in controls and patients were slightly positive for both forms of MGP, whereas Gla-MGP was more abundant within control's than within patient's elastic fibers (P<0.001). In patients' calcified elastic fibers, Glu-MGP intensively colocalized with mineral precipitates, whereas Gla-MGP precisely localized at the mineralization front. Data suggest that MGP is present within elastic fibers and is associated with calcification of dermal elastic fibers in PXE. To investigate whether local cells produce MGP, dermal fibroblasts were cultured in vitro and MGP was assayed at mRNA and protein levels. In spite of very similar MGP mRNA expression, cells from PXE patients produced 30% less of Gla-MGP compared to controls. Data were confirmed by immunocytochemistry on ultrathin sections. Normal fibroblasts in vitro were positive for both forms of MGP. PXE fibroblasts were positive for Glu-MGP and only barely positive for Gla-MGP (P<0.001). In conclusion, MGP is involved in elastic fiber calcification in PXE. The lower ratio of Gla-MGP over Glu-MGP in pathological fibroblasts compared to controls suggests these cells may play an important role in the ectopic calcification in PXE.
2007
- New insights on the pathogenesis of Pseudoxanthoma elasticum by proteome analysis.
[Abstract in Rivista]
Boraldi, Federica; Annovi, Giulia; PAOLINELLI DEVINCENZI, Chiara; Gheduzzi, Dealba; Tiozzo, Roberta; Ronchetti, Ivonne; Quaglino, Daniela
abstract
Data are presented on the protein profile of dermal fibroblasts cultured in vitro from PXE patients, with the aim to elucidate molecular pathways involved in the pathogenesis of the disease
2007
- Response of confluent human dermal fibroblasts to serum deprivation investigated by proteome analysis.
[Abstract in Rivista]
Boraldi, Federica; Annovi, Giulia; PAOLINELLI DEVINCENZI, Chiara; Guidetti, Rita; Tiozzo, Roberta; Quaglino, Daniela
abstract
A preliminary characterization of the protein profile of human dermal fibroblstats cultured in vitro after confluence, in the absence of serum factors, is provided.
2006
- Oxidative stress in fibroblasts from patients with pseudoxanthoma elasticum: possible role in the pathogenesis of clinical manifestations
[Articolo su rivista]
Ronchetti, Ivonne; MI Garcia, Fernandez; Boraldi, Federica; Quaglino, Daniela; Gheduzzi, Dealba; De Vincenzi Paolinelli, C.; Tiozzo, Roberta; Bergamini, Stefania; D., Ceccarelli; U., Muscatello
abstract
Pseudoxanthoma elasticum (PXE) is a genetic disease characterized by calcification and fragmentation of elastic fibres of the skin, cardiovascular system and eye, caused by mutations of the ABCC6 gene, which encodes the membrane transporter MRP6. The pathogenesis of the lesions is unknown. Based on studies of similar clinical and histopathological damage present in haemolytic disorders, our working hypothesis is that PXE lesions may result from chronic oxidative stress occurring in PXE cells as a consequence of MRP6 deficiency. Our results show that PXE fibroblasts suffer from mild chronic oxidative stress due to the imbalance between production and degradation of oxidant species. The findings also show that this imbalance results, at least in part, from the loss of mitochondrial membrane potential (Delta Psi(m)) with overproduction of H2O2. Whether mitochondrial dysfunction is the main factor responsible for the oxidative stress in PXE cells remains to be elucidated. However, mild chronic generalized oxidative stress could explain the great majority of structural and biochemical alterations already reported in PXE.
2005
- New insights on the pathogenesis of clinical manifestations in Pseudoxathoma elasticum.
[Abstract in Rivista]
Ronchetti, Ivonne; M. I., GARCIA FERNANDEZ; Boraldi, Federica; Quaglino, Daniela; Gheduzzi, Dealba; PAOLINELLI DEVINCENZI, Chiara; Tiozzo, Roberta; Bergamini, Stefania; Ceccarelli, Daniela; Muscatello, Umberto
abstract
Data are presented on the presence of oxidative stress in fibroblasts from PXE patients and the potential role in PXE pathogenesis is discussed.
2005
- On the pathogenesis of PXE-like clinical manifestations in Beta-thalassemic patients
[Abstract in Rivista]
Boraldi, Federica; PAOLINELLI DEVINCENZI, Chiara; M. I., Garcia Fernandez; Quaglino, Daniela; Tiozzo, Roberta; Croce, Maria Antonietta; P., Cianciulli; F., Sorrentino; G. L., Forni; Ronchetti, Ivonne
abstract
Altered fibroblast phenotype and membrane transport properties are described in patients affected by beta-thalassemia with elastic fiber mineralization and PXE-like clinical manifestations.
2004
- The skin equivalent model: developments and perspectives.
[Articolo su rivista]
Croce, Maria Antonietta; Sammarco, Rita; PAOLINELLI DEVINCENZI, Chiara; Boraldi, Federica; Gheduzzi, Dealba; Damour, O.; Sommer, P.; Ronchetti, Ivonne; Tiozzo, Roberta; Quaglino, Daniela
abstract
Three-dimensional culture systems have been developed to mimic natural interactions among cells and between cells and the extracellular matrix. The “skin equivalent ” is a 3-D co-culture system of fibroblasts and keratinocytes used as a covering surface in extended wounds, and as a model for studying the influence of each cell type on the synthesis of the extracellular matrix. In the present study, a three-dimensional scaffold made of chitosan, glycosaminoglycans and collagen has been colonised by keratinocytes from newborn foreskin an by fibroblasts isolated from donors of different ages. In the skin equivalent, the neo-synthesized connective tissue is characterized by the presence of amorphous elastic fibers, strengthening the usefulness of this three-dimensional model as a powerful tool for investigating connective tissue metabolism in different physiological (i.e. aging) and pathological conditions. Interestingly, when fibroblasts isolated from aged donors were cultured in vitro, even though in the presence of abundant nutrients and growth factors, their old phenotype appeared not to be completely overcome, indicating that aging is the result of a continuous remodelling controlled by epigenetic as well as genetic factors. Finally, the aged phenotype of fibroblasts seems to exert a stronger influence on keratinocytes compared to that of keratinocytes on fibroblasts.