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    Dexamethasone and Stanozolol affect osteogenic differentiation of SaOS-2 cells.

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    Aim: The aim of this study is to investigate the effects of Dexamethasone (DX) and Stanozolol (ST) in inducing osteogenic differentiation on SaOS-2 cells. Material and Methods: Cells were cultured in DMEM-low glucose supplemented with Fetal Bovine Serum 10%, penicillin/ streptomicin 100 lg/ml, glutammin 4 mmol/l, ascorbic acid 50 lg/ml, L-proline 260 lmol/l, b2-glicerophosphate 10 mmol/ l. Either Dexamethasone or Stanozolol at concentrations of 0, 1, 10, 100, 1000 nmol/l were furtherly added as experimental conditions. After 6, 12 and 24 days, cells were stained with Alizarin Red (AR), Von Kossa (VK) for qualitative analysis and with DAPI and calcein green for semi-quantitative analysis. Gene expression of RUNX-2 and BMP-1 was evaluated through RTPCR. Results: AR and VK stainings showed a dose-dependent mineral apposition in cells treated with ST. This finding was more evident at 12 days, while at 24 days all samples treated with ST were extensively calcified. Semi-quantitative evaluation of calcein/ DAPI confirmed a dose-dependant mineralization even at the last time-point. Samples treated with DX exhibited similar results, with a less pronounced mineral apposition. Gene expression analysis revealed a dose-dependant increase of Runx-2 in samples treated with ST compared to controls (p < 0.05), and not significant changes in DX-treated samples at any tested concentration (p > 0.05). BMP-1 expression had a significant dosedependant decrease in samples treated with DX (p < 0.05). Conclusion: Standing to our results, ST boosts osteogenic differentiation of SaOS-2 in a dose-dependent manner. Also DX may produce similar effects, at a lower rate. Further studies are required to understand steroids’ mechanism of action on osteogenic cells as well as their possible use in the field of bone regeneration

    Tailoring the Interface of Biomaterials to Design Effective Scaffolds

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    Tissue engineering (TE) is a multidisciplinary science, which including principles from material science, biology and medicine aims to develop biological substitutes to restore damaged tissues and organs. A major challenge in TE is the choice of suitable biomaterial to fabricate a scaffold that mimics native extracellular matrix guiding resident stem cells to regenerate the functional tissue. Ideally, the biomaterial should be tailored in order that the final scaffold would be (i) biodegradable to be gradually replaced by regenerating new tissue, (ii) mechanically similar to the tissue to regenerate, (iii) porous to allow cell growth as nutrient, oxygen and waste transport and (iv) bioactive to promote cell adhesion and differentiation. With this perspective, this review discusses the options and challenges facing biomaterial selection when a scaffold has to be designed. We highlight the possibilities in the final mold the materials should assume and the most effective techniques for its fabrication depending on the target tissue, including the alternatives to ameliorate its bioactivity. Furthermore, particular attention has been given to the influence that all these aspects have on resident cells considering the frontiers of materiobiology. In addition, a focus on chitosan as a versatile biomaterial for TE scaffold fabrication has been done, highlighting its latest advances in the literature on bone, skin, cartilage and cornea TE

    Modello di innesto a blocco di tipo “over-inlay” per lo studio della rigenerazione ossea nel ratto

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    Obiettivo: L’obiettivo di questo studio è di descrivere un modello di innesto a blocco per la valutazione della rigenerazione ossea nel ratto. Materiali e Metodi: Difetti critici standardizzati dal diametro di 5 mm sono stati creati nell’osso parietale di 12 ratti Wistar maschi di 4 mesi. Blocchi di osso bovino deproteinizzato sono stati inseriti all’interno dei difetti, in modo che una parte del blocco fosse alloggiata nella cavità chirurgica (parte “inlay”) ed una parte eccedesse in altezza lo spessore della teca cranica esterna (parte “over”). Sei animali sono stati sacrificati a 1 mese e 6 a 3 mesi e all’interno di ogni campione sono state identificate diverse regioni di interesse (ROI): area periostale (PA), aree laterali adiacenti all’osso nativo (BA), aree centrali (CA). In ogni ROI sono stati quantificati osso neoformato (NB) e tessuto fibroso (FT). Densità di capillari, espressione di Osterix (OSX) e collagene1 (COL1) sono state valutate con immunoistochimica. I dati sono stati analizzati tramite test 2way-ANOVA e post-test di Sidak, p ≤ 0.05. I risultati sono espressi come media ± SEM. Risultati: NB è risultato maggiore in BA rispetto a CA, a 1 mese (42183.89±6477.27 μm2, p< 0.05) e a 3 mesi (64131.11±7520.61 μm2, p<0.01). FT è risultato maggiore in PA rispetto alle altre ROI a 1 mese (PA 179192.50±29187.19 μm2, p<0.01) e 3 mesi (243367.80±55447.25 μm2, p<0.001). I dati sulla densità capillare hanno evidenziato una progressiva migrazione del processo di neoangiogenesi da BA verso CA. Positività per OSX e COL1 è stata riscontrata soprattutto in PA. Conclusioni: Il modello “over-inlay” rende possibile un approccio critico per la valutazione del contributo distinto di diverse ROI nel processo di integrazione dell’innesto: la parte inlay consente di valutare BA e CA, la parte “over,” priva dell’apporto dell’osso nativo, la PA. Questo consente di ottenere dati più facilmente interpretabili con conseguente riduzione della numerosità del modello animale
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