In the present research, titanium dioxide (TiO2) nanoparticles (NPs) were prepared by using sol-gel technique, and the phase structure and morphology of obtained TiO2 NPs were characterized by using X-ray diffraction (XRD) and scanning electron microscopy (SEM), respectively. The XRD patterns of the TiO2 NPs indicated the formation of anatase phase. A new nanocomposite film has been developed by solvent casting method from collagen hydrolysate (CH) and TiO2 NPs at different concentrations for use in packaging industry. Fourier transform infrared (FTIR) spectra showed the intermolecular reactions between CH and TiO2 NPs in the films. SEM micrographs revealed a clear dispersion of TiO2 NPs on the surface of the films especially at low concentrations. Thermal analyses showed that after incorporation of TiO2 NPs in polymer matrix, heat susceptibility of films were changed and weight loss of films was decreased. The results revealed that incorporation of TiO2 NPs significantly decremented water vapor permeability (WVP), water solubility, and elongation at break (EAB) of the films, whereas tensile strength (TS) significantly incremented. The findings pointed out that TiO2 NPs can be successfully incorporate to the CH films and the reutilization of CH in nanocomposite films will be a promising alternative material in non-food packaging industry.