LAUSR

Introduction: Mineral trioxide aggregate (MTA) regulates bone remodeling, particularly osteoclast differentiation. However, intracellular mechanisms underlying the anti‐osteoclastogenesis of MTA remain unclear. This study aimed to evaluate the potential alterations of autophagic pathway during anti‐osteoclastogenic effects by MTA in vitro and investigate their underlying mechanisms. Methods: Osteoclast precursors were treated with MTA extracts containing the receptor activator of nuclear factor‐kappa B ligand (RANKL). Rapamycin was used to activate autophagy. RANKL‐induced osteoclast differentiation was stained with tartrate‐resistant acid phosphatase. Several specific autophagy features in osteoclast precursors were measured by using immunofluorescence, monodansylcadaverine, and transmission electron microscope. Autophagy‐related proteins were investigated via Western blot analysis. The mRNA expression involved in autophagic and osteoclastic activities was detected with quantitative real‐time polymerase chain reaction. Results: MTA extracts inhibited osteoclast differentiation via preventing the fusion of osteoclast precursors without cytotoxicity. MTA extracts interrupted RANKL‐induced acidic vesicular organelle formation and autophagic vacuole appearance in osteoclast precursors. Moreover, autophagic genes and proteins stimulated with RANKL diminished with MTA extracts. Notably, autophagy activation through rapamycin promoted multinucleated osteoclasts formation and increased osteoclastic genes expression, which almost reversed MTA‐mediated anti‐osteoclastogenic effects. Conclusions: MTA inhibited osteoclastogenesis for bone repair through attenuating the autophagic pathway. HIGHLIGHTSMTA extracts inhibited autophagic LC3‐II accumulation in osteoclasts.MTA extracts downregulated formation of AVOs and AVs in osteoclast precursors.MTA extracts downregulated autophagic proteins expression in osteoclast precursorsInhibitory effects of MTA extracts on autophagy were reversed by rapamycin.MTA‐mediated anti‐osteoclastogenic effects were reversed by rapamycin.