Chitosan is an important cationic biopolymer widely used in various biomedical applications such as wound care, drug delivery, biomaterial scaffolds, and tissue engineering. In this work, hollow silica sphere (HSS) nanoparticles with an average size of 400–450 nm (via TEM) were synthesized by sol–gel process and were epoxidized using epichlorohydrin. Then, chitosan was chemically bound to the epoxidized HSS to form a bionanocomposite, HSS-epoxy-CHI-X where X represents the percentage of chitosan in the sample. BET analysis showed that pure HSS has a specific surface area (Ass) of around 4.8 m2/g and adsorption average pore diameter of 10.18 nm. Bionanocomposites were characterized by FT-IR spectroscopy to confirm the bonding of chitosan on HSS nanoparticles. X-ray diffraction studies demonstrated that the amorphous character of the materials improved with the HSS content. Thermogravimetric analysis illustrated that at low temperatures, the thermal stability of bionanocomposites was higher than that of the sample with no chitosan in it. Scanning electron microscopic analysis results confirmed the homogenous distribution of HSS in the bionanocomposites. Considering the biological activity of chitosan and absorption characteristics of HSS, newly developed HSS-epoxy-CHI-X bionanocomposites were tested for wound healing by in vitro scratch assay using NIH 3T3 fibroblast cells. Among the bionanocomposites analyzed, HSS-epoxy-CHI-60 exhibited better performance where about 90% wound closure was observed for this sample after 21 h of exposure.