Background: Osteoclasts are large terminal-differentiated cells with multiple nuclei and are the only cells with bone resorption activity in the body. The abnormal migration and differentiation of osteoclasts may accelerate bone absorption, a crucial process in the occurrence and development of osteoporosis. Regulating the differentiation and activation of osteoclasts is a breakthrough point for the prevention and treatment of osteoporosis. Coumarin derivative of 3-(4-methoxyl)-1-(2-(4-coumarin) prop)-2-en-1-one (MCPEO) was selected in this study. We aimed to investigate the effects of (MCPEO) on osteoclast differentiation.
Methods: Bone marrow mononuclear cells (BM-MNCs) were collected from 6-week-old ICR mice and inoculated in vitro. BM-MNC and RAW264.7 cells were induced by the receptor activator of nuclear factor κ B ligand (RANKL) to differentiate them into osteoclasts. The cells were then added with 0.01, 0.05, 0.1, 0.5, and 1 μM MCPEO. The cell viability of differentiated osteoclasts was analyzed by methylthiazolyldiphenyl-tetrazolium bromide assay. The differentiated osteoclasts were detected by tartrate-resistant acid phosphatase (TRAP) staining. Miuse osteoclast activation was investigated by absorptive activity analysis. Filamentous actin (F-actin) staining was employed to identify the formation of F-actin rings in the differentiated mouse osteoclasts. The change level of critical transcription factors related to osteoclast differentiation was determined by Western blot analysis.
Results: Data show that MCPEO affected the cell viability of differentiated osteoclasts, inhibited the formation of TRAP-positive polynuclear cells, and decreased the absorption activity and the formation of F-actin rings in the differentiated osteoclasts. Furthermore, MCPEO influenced the change level of crucial transcription factors related to osteoclast differentiation.
Conclusions: MCPEO inhibited the differentiation of RANKL-induced BM-MNC and RAW264.7 cells into osteoclasts.