Icariin suppresses osteogenic differentiation and promotes bone regeneration in Porphyromonas gingivalis-infected conditions through EphA2-RhoA signaling pathway.

Periodontitis is associated with multiple systemic diseases and can cause bone loss. Porphyromonas gingivalis (P. gingivalis) is one of the most virulent periodontal pathogens. Icariin is a flavonoid extracted from the traditional Chinese herbal medicine Herba Epimedii, and can regulate bone metabolism. However, its effects on promoting bone metabolism have not been fully elucidated. In this experiment, we infected MC3T3-E1 cells with P. gingivalis. Flow cytometry results show that persistent bacterial infection does not affect cell proliferative activity. Western blotting, ALP activity detection, mineral content determination, and immunofluorescence blotting confirmed that icariin improved osteogenic differentiation in the inflammatory state, and this effect may be more obvious in the early stage of osteogenic differentiation. The antibacterial assays, ROS and MMP fluorescence assays demonstrated that icariin exerted a significant inhibitory effect on bacterial growth and attenuated the inflammatory response in bacterial-infected conditions. The results of in vivo experiments in animals further validated the excellent properties exerted by icariin in the repair of bone defects. Additionally, in the P. gingivalis-infected state, icariin exert a regulatory effect on EphA2-RhoA signaling pathway to augment osteogenic differentiation. These exciting findings suggest that icariin holds significant potential for therapeutic application in the management of periodontal bone loss.