In Vitro Effects of FBS and Ovarian Steroids on Osteoblast Viability

Abstract

We evaluated the effects of different concentrations (0, 10‐8 M and 10‐7M) of progesterone (P4), testosterone (T) and estradiol (E2) on the osteoblast viability in vitro. Osteoblast lineages (OSTEO‐1) were cultured in 96‐well plates in DMEM medium a density of 5 x 103 cells/100 μL medium, with or without 10% fetal bovine serum (FBS). Cells were maintained at 37 ºC, with 5% CO2, for up to 48 hours. Cell viability was analyzed by MTT tetrazolium assay. The cells cultured with 10% FBS were more viable than those cultured without FBS. When all hormones were put together, the cell viability was reduced at the 10‐7M concentrations compared with the control and 10‐8 M. The viability of cells cultured without FBS was a) stimulated by E2 (10‐7 M and 10‐8 M (0.1048 ± 0.0227 and 0.1069 ± 0.0124, respectively)) compared with the control group (0.0566 ± 0.008); b) stimulated by P4 (10‐7 M (0.1088 ± 0.0124)) compared with the control group (0.0566 ± 0.008) and 10‐8 M (0.0924 ± 0.022); and c) stimulated by both concentrations of T (10‐7 M (0.0908 ± 0.0153) and 10‐8 M (0.0924 ± 0.0022)) compared with the control group (0.0566 ± 0.008). Our results demonstrated that FBS stimulated osteoblast viability, and P4 (10‐7 M), and both concentrations of T and E2 stimulated the viability of the cells cultured without FBS, suggesting that these steroid hormones may be important at the onset of osteoblast cells viability in vitro for up to 48 hours.