Nanotechnology offers a new strategy to develop novel bioactive materials, given that nano-scaled biomaterials exhibit an enhanced biocompatibility and bioactivity. In this work, we developed a method for the synthesis of spherical bioactive glass nanoparticles (BGNP) aimed at producing biomaterials for potential use in the repair of hard tissues. The BGNP were prepared using the sol-gel process based on the reaction of alkoxides and other precursors in aqueous media for obtaining the oxide-ternary system with the stoichiometric proportion of 60% SiO2, 36% CaO and 4% P2O5. The system was extensively characterized by Fourier transform infrared, x-ray diffraction and scanning electron microscope/energy-dispersive x-ray spectroscopy with regard to chemical composition, crystallinity and morphology. Moreover, the results suggested the BGNP to be highly bioactive, which was confirmed by the formation of a hydroxyapatite biomimetic layer on the material surfaces upon immersion in simulated body fluid solution. In addition, the bioactivity response toward the developed BGNPs was assessed by direct contact of osteoblast cells using resazurin and alkaline phosphatase assays. The new BGNP have presented a significant increase in the osteoblast in vitro cytocompatibility behavior as compared to similar micro-sized bioactive glass particles. Such improvement in the overall bioactive behavior of BGNP was attributed to the much higher surface area causing enhanced interactions at the cell-nanomaterial interfaces. Hence, based on the results, the BGNP produced are the biomaterials to be potentially utilized in hard tissue engineering applications.