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FEDERICA CAPITANI

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Dipartimento di Scienze Biomediche, Metaboliche e Neuroscienze

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Pubblicazioni

2023 - Composition and Anticoagulant Potential of Chondroitin Sulfate and Dermatan Sulfate from Inedible Parts of Garfish (Belone belone) [Articolo su rivista]
Chikha, S. B.; Bougatef, H.; Capitani, F.; Ben Amor, I.; Maccari, F.; Gargouri, J.; Sila, A.; Volpi, N.; Bougatef, A.
abstract

Glycosaminoglycans (GAGs) play a crucial role due to their significant biomedical functions. Chondroitin sulfate (CS) and dermatan sulfate (DS), the main representative family of GAGs, were extracted and purified from garfish (Belone belone) by-products, i.e., skin (GSB), bones (GCB), and heads (GHB), and their composition and anticoagulant activity were investigated. CS/DS were purified by ion-exchange chromatography with yields of 8.1% for heads, 3.7% for skin, and 1.4% for bones. Cellulose acetate electrophoresis was also explored for analyzing the extracted CS/DS. Interestingly, GHB, GSB, and GCB possessed sulfate contents of 21 ± 2%, 20 ± 1%, and 20 ± 1.5%, respectively. Physico-chemical analysis showed that there were no significant differences (p > 0.05) between the variances for sulfate, uronic acid, and total sugars in the GAGs extracted from the different parts of fish. Disaccharide analysis by SAX-HPLC showed that the GSB and GCB were predominately composed of ΔDi-4S [ΔUA-GalNAc 6S] (74.78% and 69.22%, respectively) and ΔDi-2,4S [ΔUA2S-GalNAc 4S] (10.92% and 6.55%, respectively). However, the GHB consisted of 25.55% ΔDi-6S [ΔUA-GalNAc 6S] and 6.28% ΔDi-2,6S [ΔUA2S-GalNAc 4S]. Moreover, classical anticoagulation tests were also used to measure their anticoagulant properties in vitro, which included the activated partial thromboplastin time, prothrombin time, and thrombin time. The CS/DS isolated from garfish by-products exhibited potent anticoagulant effects. The purified CS/DS showed exceptional anticoagulant properties according to this research and can be considered as a new agent with anticoagulant properties.

2020 - Anti-zika virus and anti-usutu virus activity of human milk and its components [Articolo su rivista]
Francese, R.; Civra, A.; Donalisio, M.; Volpi, N.; Capitani, F.; Sottemano, S.; Tonetto, P.; Coscia, A.; Maiocco, G.; Moro, G. E.; Bertino, E.; Lembo, D.
abstract

The benefits of human milk are mediated by multiple nutritional, trophic, and immunological components, able to promote infant’s growth, maturation of its immature gut, and to confer protection against infections. Despite these widely recognized properties, breast-feeding represents an important mother-to-child transmission route of some viral infections. Different studies show that some flaviviruses can occasionally be detected in breast milk, but their transmission to the newborn is still controversial. The aim of this study is to investigate the antiviral activity of human milk (HM) in its different stages of maturation against two emerging flaviviruses, namely Zika virus (ZIKV) and Usutu virus (USUV) and to verify whether HM-derived extracellular vesicles (EVs) and glycosaminoglycans (GAGs) contribute to the milk protective effect. Colostrum, transitional and mature milk samples were collected from 39 healthy donors. The aqueous fractions were tested in vitro with specific antiviral assays and EVs and GAGs were derived and characterized. HM showed antiviral activity against ZIKV and USUV at all the stages of lactation with no significant differences in the activity of colostrum, transitional or mature milk. Mechanism of action studies demonstrated that colostrum does not inacti-vate viral particles, but it hampers the binding of both flaviviruses to cells. We also demonstrated that HM-EVs and HM-GAGs contribute, at least in part, to the anti-ZIKV and anti-USUV action of HM. This study discloses the intrinsic antiviral activity of HM against ZIKV and USUV and demonstrates the contribution of two bioactive components in mediating its protective effect. Since the potential infectivity of HM during ZIKV and USUV infection is still unclear, these data support the World Health Organization recommendations about breast-feeding during ZIKV infection and could contribute to producing new guidelines for a possible USUV epidemic.

2020 - Short-and long-term effectiveness of supplementation with non-animal chondroitin sulphate on inflammation, oxidative stress and functional status in obese subjects with moderate knee osteoarthritis before and after physical stress: A randomized, double-blind, placebo-controlled trial [Articolo su rivista]
Rondanelli, M.; Miraglia, N.; Putignano, P.; Peroni, G.; Faliva, M. A.; Naso, M.; Gasparri, C.; Infantino, V.; Nichetti, M.; Volpi, N.; Capitani, F.; Mantovani, V.; Perna, S.
abstract

It has recently been demonstrated that chronic supplementation with nonanimal chondroitin sulfate (nonanimal CS) in overweight subjects with knee osteoarthritis (OA) improves the function, pain and inflammation, but there are no studies of its effectiveness in an acute setting. In 48 obese subjects with moderate knee OA, we investigated the effectiveness of nonanimal CS supplementation for eight weeks on the inflammation, functional status, oxidative stress, cartilage catabolism markers, metabolic profile and body composition, by Dual-Energy X-ray Absorptiometry (DXA) at the baseline, after 15 days and at the end of the eight-week study. To evaluate the acute effectiveness on inflammation, 15-min cycle training sessions were done 15 days after the start of the study and at the end. C-reactive protein (CRP) was assayed in blood samples collected before and after the two cycling exercises. The 48 obese subjects (M and F, 20–50 years, body mass index (BMI) 30–35 kg/m2) were randomly assigned to an experimental group (N = 24, 600-mg tablet of nonanimal CS/day) or the control group (N = 24, placebo). The between-groups analysis of covariance showed a significant effect on the Western Ontario and McMaster Universities Arthritis index (WOMAC) scale (p = 0.000) and CRP (p = 0.022). For intra-group differences, the result was significant in the CS group for BMI, WOMAC, CRP, total cholesterol and Homeostasis Model Assessment (HOMA). In these obese adults with OA, nonanimal CS improved the inflammation, knee function, metabolic profile and body composition.

2020 - Structural definition of terrestrial chondroitin sulfate of various origin and repeatability of the production process [Articolo su rivista]
Volpi, N.; Galeotti, F.; Maccari, F.; Capitani, F.; Mantovani, V.
abstract

We report results on the structure, physicochemical characteristics and purity of chondroitin sulfate (CS) samples derived from three largely available and common biological sources such as bovine and porcine trachea and chicken keel bones with the aim to define their structural signatures. Many lots of CS produced by a manufacturer at industrial scale were characterized with a view to assess the reproducibility of the process as not controlled extractive procedures may produce final products with variable structure and biological contaminants as well as not constant clinical efficacy and safety. By using standardized source animal tissues and manufacturing procedure, highly pure CS (∼92 %) products with constant structure and characteristics were obtained. Bovine CS showed a lower molecular weight (MWw of ∼21,500 Da) than porcine (MWw of ∼26,000 Da) and chicken (MWw of ∼35,900 Da) products with a CV% of ∼2.0–7.5 and a polydispersity variability of 0.7–2.7 %. The ratio between the sulfate groups main located in position 4 and 6 of N-acetyl-galactosamine (4/6 ratio) was ∼1.70 for bovine CS versus a value of 3.60 for porcine and ∼2.70 for chicken samples with a overall charge density of 0.92−0.93 and a CV% of 2.1−2.5. The final products also showed the presence of a very low and constant content of other co-purified bio(macro)molecules (hyaluronic acid, keratan sulfate, dermatan sulfate, heparan sulfate, nucleic acids and proteins), calcium and sodium, and the absence of versican. Finally, a high reproducibility of molecular weight values, disaccharide composition, specific optical rotation and particle dimension was observed. The observed parameters are structural signatures useful to specifically identify the origin of CS and obtained by a standardized and highly reproducible manufacturing process. The compositional profile determined from this study provides a measure of the norm and range of variation in CS samples of terrestrial origin produced under standardized production protocol to which future pharmaceutical/nutraceutical final products can be compared. Moreover, the physicochemical properties including molecular weight, disaccharide composition, presence of natural contaminants and particle dimension were characterized to provide the basis of CS of high quality for application as pharmaceutical/nutraceutical active agents.

2019 - Purification, compositional analysis, and anticoagulant capacity of chondroitin sulfate/dermatan sulfate from bone of corb (Sciaena umbra). [Articolo su rivista]
Bougatef, H; Krichen, F; Capitani, F; Amor, Ib; Gargouri, J; Maccari, F; Mantovani, V; Galeotti, F; Volpi, N; Bougatef, A; Sila, A.
abstract

Chondroitin sulfate/dermatan sulfate (CS/DS) were isolated and purified for the first time from the bone of corb (Sciaena umbra) (CBG) and their chemical composition and anticoagulant activity were assessed. Infrared spectrum and agarose-gel electrophoresis for extracted CS/DS were also investigated. The results showed that the purified CS/DS obtained at a yield of 10% contains about 31.28% sulfate and an average molecular mass of 23.35 kDa. Disaccharide analysis indicated that CBG was composed of monosulfated disaccharides in positions 6 and 4 of the N-acetylgalactosamine (8.6% and 40.0%, respectively) and disulfated disaccharides in different percentages. The charge density was 1.4 and the ratio of 4:6 sulfated residues was equal to 4.64. Chondroitinase AC showed that the purified CS/DS contained mainly 74% CS and 26% DS. Moreover, the new CS/DS extracted from bone of corb showed a strong anticoagulant effect through activated partial thrombosis time (aPTT), thrombin time (TT) and prothrombin time (PT). In fact, CBG prolonged significantly (p < 0.05), aPTT and PT about 2.62 and 1.26 fold, respectively, greater than that of the negative control at a concentration of 1000 μg/mL. However, TT assay of CBG was prolonged 3.53 fold compared with the control at 100 μg/mL. The purified CS/DS displayed a promising anticoagulant potential, which may be used as a novel and soothing drug.

2019 - Recent advances in analytical approaches for the standardization and quality of polyphenols of propolis [Articolo su rivista]
Galeotti, F; Capitani, F; Fachini, A; Volpi, N.
abstract

Analytical approaches utilized for the characterization of polyphenols from propolis useful for the determination of its quality is investigated in this study. A qualitative and quantitative evaluation of propolis bioactive molecules is of interest in medicine and nutraceuticals. Recent powerful analytical techniques are of great utility to separate and quantify polyphenols in extracts and finished products due to their capacity to produce typical fingerprints and a reliable identification of many components. According to this, an HPLC-UV-MS procedure was validated and applied for the characterization and quantification of bioactive substances in propolis and for an accurate assessment of their content in extract samples. By using this analytical approach, we obtained specific compositions related to brown propolis acquired from different geographic areas (and preparations and treatment). This is more important by considering the scientific opinion of European Food Safety Authority (EFSA) which provided a negative response related to health claims of propolis and its polyphenols. These results prove that HPLC-MS is an attractive tool for the standardization and quality control of propolis and may be realistically applied to screen raw material and to evaluate finished commercial preparations and nutraceutical benefits.

2018 - Chondroitin sulfate/dermatan sulfate from corb (Sciaena umbra) skin: Purification, structural analysis and anticoagulant effect [Articolo su rivista]
Bougatef, H; Krichen, F; Capitani, F; Amor, Ib; Maccari, F; Mantovani, V; Galeotti, F; Volpi, N; Bougatef, A; Sila, A.
abstract

In this study, chondroitin sulfate/dermatan sulfate was isolated and purified from the skin of corb (Sciaena umbra) (CSG) with a yield of 6.2%. Chemical and structural analysis showed that CSG consisted of high sulfate content 28.74% and an average molecular weight of 15.46 KDa. The separation of CSG by agarose-gel electrophoresis revealed the presence of DS and CS. Structural analysis of the purified CS/DS by means of SAX-HPLC after treatment with specific chondroitinases showed that this polymer was composed of nonsulfated disaccharide, monosulfated disaccharides and disulfated disaccharides in various percentages. The results also suggest that the percentage of CS and DS recovred in CSG were 24% and 76%, respectively. Anticoagulant activity in vitro was measured in plasma using classical anticoagulation tests: activated partial thromboplastin time (aPTT), thrombin time (TT) and prothrombine time (PT) tests. The findings thus indicated that the purified CS/DS exhibits a remarkably high anticoagulant effect.

2018 - Isolation, Purification and Structural Characterestics of Chondroitin Sulfate from Smooth hound Cartilage: In vitro Anticoagulant and Antiproliferative Properties [Articolo su rivista]
Krichen, F; Bougatef, H; Sayari, N; Capitani, F; Amor, Ib; Maccari, F; Mantovani, V; Galeotti, F; Volpi, N; Bougatef, A.
abstract

Chondroitin sulfate was extracted from the cartilage of smooth hound (CSSH) and then purified by anion exchange chromatography. The structual characteristic of CSSH was evaluated by acetate cellulose electrophoresis, FTIR, 13C NMR and SAX-HPLC. Molecular weight of CSSH was average 68.78 KDa. Disaccharide analysis indicated that CSSH was predominately composed of monosulfated disaccharides in position 6 and 4 of the N-acetylgalactosamine (45.34% and 32.49%, respectively). CSSH was tested for in vitro anticoagulant activity using the three classical coagulation assays (activated partial thromboplastin time (aPTT), prothrombine time (TT) and thrombin time (PT) tests). The finding showed that CSSH prolonged significatively (p < 0.05), aPTT, TT and PT about 1.4, 3.44 and 1.21 fold, respectively, greater than that of the negative control at a concentration of 100 μg/ml. The CSSH caused a significant antiproliferative activity against HCT116 cell, which was 79% of cell proliferation inhibition at the concentration of 1000 μg/ml. Further, CSSH presented no toxicity against the normal cells and no hemolysis towards bovine erythrocytes for all concentrations tested. CSSH demonstrated hopeful antiproliferative and anticoagulant potential, which may be used as a novel and effective drug.

2018 - Purification and structural elucidation of chondroitin sulfate/dermatan sulfate from Atlantic bluefin tuna (Thunnus thynnus) skins and their anticoagulant and ACE inhibitory activities [Articolo su rivista]
Fatma, Krichen; Hajer, Bougatef; Capitani, Federica; Ikram Ben Amor, ; Imed, Koubaa; Jalel, Gargouri; Maccari, Francesca; Mantovani, Veronica; Galeotti, Fabio; Volpi, Nicola; Ali, Bougatef
abstract

Chondroitin sulfate/dermatan sulfate (CS/DS) was extracted from Atlantic bluefin tuna (Thunnus thynnus) skin (SGAT) and was purified and characterized. SGAT was characterized by acetate cellulose electrophoresis, FTIR spectroscopy, 13C NMR spectroscopy and SAX-HPLC. According to the results obtained for specific chondroitinases (ABC and AC) and the SAX-HPLC separation of generated unsaturated repeating disaccharides, the polymer was found to contain a disaccharide monosulfated in positions 6 and 4 of GalNAc and disulfated disaccharides in different percentages. These results were confirmed by 13C NMR experiments. The average molecular mass was 24.07 kDa, as determined by PAGE analysis. SGAT was evaluated for its in vitro anticoagulant activity via activated partial thromboplastin time, thrombin time and prothrombin time tests. The polymer showed strong inhibitory activity against angiotensin I-converting enzyme (IC50 ¼ 0.25 mg mL1). Overall, the results suggest that this newly extracted CS/DS can be useful for pharmacological applications.

2018 - Studies on European eel skin sulfated glycosaminoglycans: Recovery, structural characterization and anticoagulant activity [Articolo su rivista]
Sila, A; Bougatef, H; Capitani, F; Krichen, F; Mantovani, V; Amor, Ib; Galeotti, F; Maccari, F; Nedjar, N; Volpi, N; Bougatef, A.
abstract

The goal of the present work was thè extraction and structural characterization of novel sulfated glycosaminoglycans from European eel skin. The recovered glycosaminoglycans were physicochemically characterized and thè uronic acid and sulfate contents were 35.12±2.13% and 16.32±0.4%, respectìvely. Cellulose acetate electrophoresis for extracted glycosaminoglycans was also investigated. Molecular weightof these sulfated glycosaminoglycans was determined (-37 kDa) by the gradient PAGE. Glycosaminoglycans obtained from thè European eel were composed of non-sulfated, mono- and disulfated disaccharides. These sulfated glycosaminoglycans were evaluated for their in vitro anticoagulant activity using activated partial thromboplastin time, thrombin time and prothrombin time tests. The result showed that thè recovered glycosaminoglycans exhibited interestingly anticoagulant activity. These glycosaminoglycans did not show haemolytic activity towards human erythrocytes. Furthermore, these bioactive substances can be explored as a functional food with antithrombotic function or used as source of anticoagulant drugs.

2016 - A Novel Forensic Investigation Applied to Bone Remains Exhumed near to Quirra Interforce Firing Range [Articolo su rivista]
Roncati, L.; Gatti, A. M.; Capitani, F.; Bonacorsi, G.; Barbolini, G.; Maiorana, A.
abstract

The need to implement novel techniques, able to support a causal link between exposure and pathology, has been emerged over the recent years. The application of scanning electron microscope coupled with probe X-ray microanalysis (by means of an energy-dispersive spectroscopy) has been developed by our research group for the bone remains investigation. It was aimed to testify the exposure to microsized and nanosized pollutions, due to military activities in the Quirra interforce firing range, of a Sardinian shepherd, died of acute leukemia. Metallic debris with a combustive morphology and with an oncogenic potential has been surely detected inside his bone marrow canal. This novel technique has proved to be able to bring to light a source of past exposure preserved over time within the bone marrow canal. It can be useful for postmortem analyses, delivering a new avant-garde approach to modern forensic science.