Three-dimensional porous hydrogel films made of gelatin (Gel) and poly(vinyl alcohol) (PVA) were developed using the freeze-thaw method, incorporating varying concentrations (1%, 3%, and 5%) of titanium dioxide nanoparticles (TiO2-NPs) for potential wound dressing applications. The characterization of these films involved multiple techniques, including scanning electron microscopy and Fourier transform infrared spectroscopy. Key properties such as swelling degree, degradation behavior, water vapor transmission rate (WVTR), water contact angle (WCA), mechanical strength, biocompatibility, and in vivo wound healing efficacy were thoroughly analyzed. The observed differences were statistically significant (p < 0.05), confirming the reliability of the improvements. Among the formulations, the Gel/PVA composite with 3% TiO2-NPs exhibited superior mechanical properties, including a tensile strength of 15.56 MPa, elongation at break of 29.16%, and Young's modulus of 288.7 MPa (p < 0.05). This formulation also demonstrated an optimal 3D network structure, high WVTR (2546.67 g/m(2)/day), moderate WCA (67.63 degrees), and a degradation rate of 29.4%. The Gel/PVA/3% TiO2-NPs film showed enhanced compatibility with fibroblast cells and improved wound healing efficacy in vivo compared to the Gel/PVA film alone (p < 0.05). The observed biological performance is attributed to the ability of TiO2-NPs to enhance cell adhesion and proliferation via improved surface interactions and moderate reactive oxygen species modulation. While previous studies have incorporated TiO2-NPs into hydrogels, few have systematically compared their physicochemical and biological effects across multiple concentrations in a single formulation. Our findings demonstrate that the 3% TiO2-NPs composite uniquely balances mechanical strength, degradation rate, and biological efficacy. This study highlights the potential of TiO2-reinforced Gel/PVA hydrogels to address key limitations of conventional wound dressings, offering a well-balanced solution in terms of structural integrity, controlled degradation, and enhanced healing response.