Liensinine-loaded extracellular vesicles mitigate osteoporosis through inhibition of NF-κB signaling and calcium oscillations in osteoclasts.

Osteoporosis is a prevalent chronic bone disease characterized by excessive osteoclast-mediated bone resorption. Liensinine (LIE), a plant-derived alkaloid, has shown anti-inflammatory potential but its role in bone resorption remains rarely investigated. This study aimed to predict the anti-osteoclastogenic mechanism of LIE via network pharmacology and evaluate the therapeutic potential of endothelial cell-derived extracellular vesicles loaded with LIE (LIE@EVs). In this study, extracellular vesicles derived from endothelial cells were isolated, identified, and utilized to encapsulate LIE. The physicochemical properties and internalization of LIE@EVs were assessed. Core pathways of LIE were predicted using network pharmacology and molecular docking. Furthermore, both in vitro and in vivo experiments were conducted to evaluate its inhibitory effects on RANKL-induced osteoclastogenesis and bone resorption. LIE@EVs exhibited characteristics of extracellular vesicles and enhanced delivery of LIE to osteoclasts. Functional assays demonstrated that LIE@EVs significantly inhibited osteoclast differentiation and bone resorption compared to free LIE. Network pharmacology identified the NF-κB and calcium signaling pathways as the potential targets of LIE. Mechanistically, LIE@EVs effectively suppressed the activation of the NF-κB signaling pathway and calcium oscillations, ultimately downregulating osteoclast-specific gene expression. These findings exhibit the potential therapeutic significance of LIE@EVs in bone loss diseases and warrant further exploration in clinical applications for osteoporosis treatment.