The aim of the present study was to evaluate and compare the most common parameters that characterize the expression of primary osteoblast cultures from different origin (human, rat, sheep), and of the human osteosarcoma cell line MG-63 before and after stimulation with vitamin 1,25(OH)2D3. Cell viability was quite similar for primary osteoblast cultures (MTT: 1.64-2.11 OD); a significant (P <0.005) difference was found between sheep osteoblasts and MG-63 (ΔMTT: 0.52 ± 0.20 OD). Osteocalcin synthesis ranged from 15.18 to 27.00 pg/ml in primary osteoblast cultures, while it was significantly (P <0.01) lower in MG-63 (OC: 6.67 ± 0.52 pg/ml) when compared with primary human osteoblasts. Alkaline phosphatase, C-terminal procollagen type I, and interleukin-6 were significantly (P <0.005) lower in rat osteoblasts when compared with primary human osteoblasts, and similarly transforming growth factor-β1 was significantly (P <0.05) lower in rat and sheep osteoblasts when compared with primary human osteoblasts and MG-63. Nitric oxide synthesis did not show any significant difference either before or after vitamin 1,25(OH)2D3 stimulation. In conclusion, the current findings confirm the presence of interspecies differences between the selected osteoblast lineages before and after stimulation with vitamin1,25(OH)2D3. Above all, the culture of sheep osteoblasts was seen to behave more similarly to that of primary human cells, mainly in terms of cell viability, osteocalcin, interleukin-6 and transforming growth factor-β1 production.
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