Figure 2
Inhibitory effects of HRT-BU and fHRT-BU on osteoclast formation in BMMs and BMCs-osteoblast coculture. (A) BMMs were incubated with M-CSF (60 ng/ml) and the indicated concentrations of HRT-BU or fHRT-BU for 3 days, and then cell viability of BMMs was measured by CCK-8 assay. (B) BMMs were incubated with M-CSF, RANKL (150 ng/ml), and the indicated concentrations of each sample for 4 days. (C) BMCs and primary osteoblasts were cocultured with the indicated concentrations of each sample and 1,25(OH)2D3 (1 × 10-8 M) for 6 days. The cells were stained for TRAP activity and the number of TRAP-positive multinuclear osteoclasts (TRAP(+)MNCs) was counted. (D) Representative microscopic pictures of RANKL-induced multinuclear osteoclasts were shown (100×). Data are mean ± SD (n = 3). a P < 0.05, versus vehicle. b P < 0.05, versus HRT-BU.