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Bone repair bionanocomposite scaffolds were produced by incorporating dense bioactive glass nanoparticles or mesoporous bioactive glass nanospheres into a chitosan-gelatin polymer blend. The in vitro bioactivity of the scaffolds was assessed in simulated body fluid, and cell viability and osteogenic differentiation assays were performed with dental pulp stem cells. Bone regeneration properties of the scaffold materials were in vivo assessed by using a critical-sized femoral defect model in rat. The scaffold nanocomposites showed excellent cytocompatibility and ability to accelerate the crystallization of bone-like apatite in vitro. Bionanocomposites prepared with bioactive glass nanoparticles were particularly more active to promote the osteogenic differentiation of dental pulp stem cells as judged by the higher activity of alkaline phosphatase. This result is attributed to the faster dissolution of bioactive glass nanoparticles into osteogenic ionic products compared to mesoporous bioactive glass nanospheres. In vivo experiments demonstrated that bioactive glass nanoparticles (5%)/chitosan-gelatin bionanocomposite significantly produces the highest amount of new bone (∼80%) in the defect area after eight weeks of implantation. The bone regeneration capacity exhibited by the scaffolds formulated with nanodimensional bioactive glass particles make them attractive for bone reconstruction applications.