Common materials used in tissue engineering are not cost-effective and natural origin. Hence, we designed green, safe, and antibacterial bionanocomposite (bio-NC) films based on polysaccharides, chitosan (CS) and gum tragacanth (GT) for the bone tissue engineering. For this aim, different percentages of titanium dioxide nanoparticles (TiO2 NPs) and green silver (Ag)-doped TiO2 NPs (TiO2@Ag hybrid) were employed as nanofiller to improve the properties of the CS-GT film. Moreover, the physicochemical characteristics of the bio-NC films were examined with a field-emission scanning electron microscope (FE-SEM), Fourier transform infrared, X-ray diffraction, and transmission electron microscopy (TEM). The FE-SEM images showed a rough surface for the CS-GT/TiO2@Ag bio-NC films, and TEM images showed better dispersion of TiO2@Ag hybrid than TiO2 NPs into the CS-GT matrix. Also, these bio-NC films were considered for the bioactivity test and the results showed formation of hydroxyapatite on the surface of the prepared bio-NC films. Furthermore, addition of GT led to an increase in the bioactivity of the CS-GT blend. Finally, antibacterial behavior of the prepared bio-NC films was investigated against Escherichia coli and Staphylococcus aureus bacteria with/without ultraviolet irradiation and the results indicated better antibacterial performance for the CS-GT/TiO2@Ag bio-NC film (TiO2:Ag = 1:1) under both conditions.