Engineered bone-targeting Lycium barbarum L.-derived extracellular vesicle-like nanoparticles deliver miR167a-5p to regulate mitochondrial dynamics and metabolic reprogramming in alleviating osteoporosis.

With the exacerbation of global population aging, the development of novel therapeutic strategies for osteoporosis (OP) has emerged as a pressing worldwide challenge. As an emerging natural nanomedicine, medicinal plant-derived extracellular vesicle-like nanoparticles (EVLPs) have shown considerable potential in OP treatment due to their advantages, such as good bioavailability, high biosafety characteristics, and natural targeting. Herein, we isolated Lycium barbarum L.-derived EVLPs (LB-EVLPs) from fresh LB via ultracentrifugation combined with sucrose gradient centrifugation and further functionalized LB-EVLPs with the bone-targeting peptide SDSSD (BT-LB-EVLPs) to treat OP. This engineered LB-EVLPs exhibited selective targeting of bone tissue and were effectively internalized by bone marrow mesenchymal stem cells (BMSCs), with a distinct propensity for mitochondrial localization. In ovariectomized (OVX)-induced osteoporotic mice, BT-LB-EVLPs alleviated bone loss, improved bone microstructure, and enhanced bone strength. Proteomic profiling indicated that BT-LB-EVLPs reprogram mitochondrial metabolism by enhancing oxidative phosphorylation while suppressing excessive glycolytic flux, thereby exerting anti-osteoporotic effects. In vitro experiments demonstrated that BT-LB-EVLPs attenuated oxidative stress, promoted mitochondrial fusion, inhibited mitochondrial fission, and facilitated metabolic reprogramming in BMSCs, ultimately restoring mitochondrial function and enhancing osteogenic differentiation. Through lentiviral-mediated overexpression of SLC25A26 combined with miR167a-5p mimic/inhibitor interventions, we verified that miR167a-5p derived from BT-LB-EVLPs directly targets the mitochondrial transporter gene SLC25A26, thereby regulating mitochondrial dynamics, sustaining energy metabolism balance, and promoting osteoblastogenesis. Additionally, in vivo knockdown of miR167a-5p exacerbated bone loss and bone microstructural damage, and abolished the anti-osteoporosis effect of BT-LB-EVLPs. Collectively, these findings emphasized this engineered LB-EVLPs as a promising targeted nanotherapeutic approach for OP treatment.