Figure 2

Effects of HG/SMG on the osteogenic differentiation of rBMSCs. rBMSCs were cultured under HG/SMG conditions with or without osteoblastic inducement medium for 1, 3, 5 or 7 days during a culture period of 14 days. RT-PCR analysis for ALP, cbfa1 and ColIA1 mRNA expressions:(A) Electrophoresis graph of PCR products, B) Gray intensity analysis of electrophoresis bands. (C) Western-blot analysis of Cbfa1 expression. OS indicates a positive control (RNA extracted from ROS17/2.8 cells); Cont, cells were cultured under normal conditions without inducer; O, rBMSCs were added with inducer for osteoblast; H, HG culture; M, SMG culture; numbers 1, 3, 5, 7 indicate the days of HG/SMG culture. Take samples: OH3, cells were cultured under HG for 3 day with osteogenic inducer; M7, cells were cultured under SMG for 7 days without osteogenic inducer. SMG-/HG-, SMG/HG treated without inducer; SMG+/HG+, SMG/HG treated with inducer. * p < 0.05 and ** P < 0.01, compared with relative 0 d group (Cont, O) using student t-test analysis(n = 3). The experiments were conducted twice. The GAPDH house-keeping gene was used as a control. HG increased the expression of ALP, Cbfa1 and ColIA1 in rBMSCs, whereas SMG decreased the expression levels.