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

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.