Programmed release of vascular endothelial growth factor and exosome from injectable chitosan nanofibrous microsphere-based PLGA-PEG-PLGA hydrogel for enhanced bone regeneration.

The healing of large bone defects remains a significant challenge in clinical practice. Accelerating both angiogenesis and osteogenesis can promote effective bone healing. In the natural healing process, angiogenesis precedes osteogenesis, providing a blood supply that supports the subsequent progression of osteogenesis. Developing a biomimetic scaffold that mimics the in vivo environment and promotes the proper sequence of vascularization followed by ossification is crucial for successful bone regeneration. In this study, a novel injectable dual-drug programmed releasing chitosan nanofibrous microsphere-based poly(D, l-lactide-co-glycolide)-b-poly(ethylene glycol)-b-poly(D,l-lactide-co-glycolide) (PLGA-PEG-PLGA) hydrogel is fabricated by incorporating vascular endothelial growth factor (VEGF) and microspheres loaded with dental pulp stem cells-derived exosomes (DPSCs-Exo). Rapid release of VEGF promotes the swift initiation of angiogenesis, while DPSCs-Exo release ensures persistent osteogenesis. Our results demonstrate that chitosan microsphere-based PLGA-PEG-PLGA hydrogel significantly promotes angiogenesis in human umbilical vascular endothelial cells and enhances the osteogenic differentiation of pre-osteoblasts. Furthermore, in vivo transplantation of this injectable chitosan microsphere-based PLGA-PEG-PLGA hydrogel into calvarial bone defects markedly promotes bone formation. Overall, our study provides a promising approach for improving bone regeneration by temporally replicating the behavior of angiogenesis and osteogenesis.