| Objective:To investigate the effect of NBD peptide on RANKL-stimulated osteoclastogenesis and bone-resorbing activity.Methods:It was designed of three groups, including vehicle (DMSO)control, NBD1(10uM) and NBD2(20uM). Murine preosteoclast cell (RAW264.7) proliferation was monitored using (AB) cell viability viability reagent and, effects of NBD peptide on RANKL-stimulated osteoclastogenesis were monitored using tartrate-resistant acid phosphatase (TRAP) activity and and the number of TRAP positive multinucleated osteoclasts (TRAP+MNCs), relative gene expressions. Resorption pits on bone slice formed by active osteoclasts were observed by scanning electron microscope.Results:Compared to control group, there were significant changes in NBD1(0.77±0.069) and NBD2(0.68±0.104) groups in tartrate-resistant acid phosphatase (TRAP) activity. The numbers of TRAP+MNCs in the two NBD groups were192±10.58(NBD1),169±8.54(NBD2), which were significantly reduced compared to235.67±12.66(DMSO). Trap gene expressions were significantly reduced in NBD1(0.77±0.069) and NBD2(0.68±0.104) compared to control group.Cathepsin K gene expressions were also significantly reduced in NBD1(0.907±0.039) and NBD2 (0.854±0.041) compared to control group. Calcitonin receptor gene expressions were increased in NBD1(1.322±0.187) and NBD2(1.498±0.128) compared to control group, and there was significant change between NBD2and control group. Proliferation of preosteoclast cell (RAW264.7) showed no significant changes between all groups. The pits on bone slice were observed by scanning electron microscope (SEM), and the resorption pits in NBD groups were visibly reduced.Conclusion:Our results indicated that NBD peptide inhibited RANKL-stimulated osteoclastogenesis from RAW264.7and bone-resorbing activity in vitro. |
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