Osteogenic human MSC-derived extracellular vesicles regulate MSC activity and osteogenic differentiation and promote bone regeneration in a rat calvarial defect model

• Osteo-EVs derived from hMSCs during early osteogenesis are enriched with bone-related proteins and exhibit enhanced osteoinductive properties compared to Naïve-EVs. • Both Naïve-EVs and Osteo-EVs promote hMSC proliferation and migration, but Osteo-EVs significantly upregulate osteogenic gene expression and calcium deposition in vitro. • In a rat calvarial defect model, Osteo-EVs combined with collagen membrane scaffold led to superior and more consistent bone regeneration than scaffold alone. • The study highlights the importance of microenvironmental priming of MSCs to produce EVs with enhanced therapeutic potential for bone tissue engineering.