Abstract

STUDY DESIGN An in vitro biological study. OBJECTIVE To explore the role of bone morphogenetic protein-2 (BMP-2) in the regulation of osteoclast-mediated osteolysis, and the possible mechanism involving BMP-2 and Nuclear factor-kappa B (NF-κB) signaling pathways. SUMMARY OF BACKGROUND DATE Recombinant human BMP-2 (rhBMP-2) has been approved as a therapeutic agent in spinal fusion and bone defect repair. However, its efficacy and clinical application are limited by associated complications including osteoclast-mediated bone resorption. The mechanism of BMP-2-induced osteolysis remains unknown. METHODS Bone marrow-derived macrophages (BMMs) were isolated from C57BL/6J mice and cultured with Macrophage Colony Stimulating Factor (M-CSF) and Receptor Activator for Nuclear Factor-κB Ligand (RANKL) to induce osteoclast differentiation. An in vitro bone resorption assay was performed by co-culturing BMMs and bone slides. The expression of BMP canonical and NF-κB signaling factors and their interaction during signal transduction were quantitated by RT-PCR, Western blot analysis, confocal microscopy and co-immunoprecipitation. RESULTS BMP-2 enhanced osteoclast-mediated bone resorption via inducing osteoclast differentiation in a concentration-dependent manner. In addition, a high concentration of BMP-2 significant up-regulated phosphorylation of BMP signaling factors p-Smad1/5/8 and NF-κB downstream factor p65, and promoted the degeneration of IκBα. In addition, BMP-2 induced osteoclast differentiation through coupling between BMP receptor II and RANK. CONCLUSIONS High concentrations of BMP-2 enhanced osteoclast-mediated bone resorption by promoting RANKL-induced pre-osteoclast differentiation, probably by mediating the cross talk between BMP canonical and NF-κB signaling pathways. LEVEL OF EVIDENCE N/A.