Injectable Gel Constructs with Regenerative and Anti-Infective Dual Effects Based on Assembled Chitosan Microspheres

There is increasing demand for biomaterials that both assist with bone regeneration and have anti-infection qualities in clinical applications. To achieve this goal, chitosan microspheres with either positive or negative charges were fabricated and then assembled as a gel for bone healing. The positively charged chitosan microspheres (CSM; ∼35.5 μm) and negatively charged O-carboxymethyl chitosan microspheres (CMCSM; ∼13.5 μm) were loaded, respectively, with bone morphogenetic protein (BMP-2) and berberine (Bbr) via swollen encapsulation and physical adsorption without a significant change in the electric charges. The release kinetics of BMP-2 and Bbr from the microspheres were also studied in vitro. The results showed that the Bbr/CMCSM microsphere group possessed high antibacterial activity against Staphylococcus aureus; the BMP-2/CSM microsphere group also had excellent cytocompatibility and improved osteoinductivity with the assistance of BMP-2. The assembled gel group consisting of Bbr/CMCSM and BMP-2/CSM had a porous structure that allowed biological signal transfer and tissue infiltration and exhibited significantly enhanced bone reconstruction compared with that of the respective microsphere groups, which should result from the osteoconductivity of the porous structure and the osteoinduction of the BMP-2 growth factor. The oppositely charged microspheres and their assembled gel provide a promising prospect for making injectable tissue-engineered constructs with regenerative and anti-infective dual effects for biomedical applications.