This laboratory study aimed to synthesize and characterize various formulations of polyvinyl alcohol (PVA)-based hydrogels by incorporating 1.0 and 3.0 wt% of hyaluronic acid (HA) and zinc oxide (ZnO) nanoparticles (NPs) for skin wound healing. Six distinct PVA-HA based hydrogel formulations were developed with varying concentrations of ZnO (1 wt% and 3 wt%) using a freeze-thaw method comprising four cycles at −22 °C. The formulations were characterized through Fourier Transform Infrared Spectroscopy (FTIR), degradation analysis, moisture content analysis, pH sensitivity analysis, tensile strength testing, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and cell behavior assessment. Data were analyzed using one-way and two-way analysis of variance with a significance threshold set at 0.05. FTIR spectra confirmed functional groups that enhance the biocompatibility, mechanical stability, and hydrophobicity of the hydrogels. Among all the formulations, PVA-HA-ZnO 1 % demonstrated adequate durability and responsiveness to different pH conditions. The experimental hydrogels exhibited swelling in acidic environments, while shrinkage was observed in basic medium. A p-value of 0.005 in pH 6.5 vs 7.5 and a p-value of 0.0006 in pH 6.7 vs 8.1 was calculated. A significant difference in degradation characteristics was evident when HA was added to PVA (p-value of 0.04) and also when pure PVA was compared with samples containing both the ZnO and HA in conjugate (p-value of 0.03 for PVA vs PVA-HA-Zn01 % and p-value of 0.04 for PVA vs PVA-HA-ZnO3 %). Thermal stability testing indicated that increasing the concentration of ZnO NPs enhanced the thermal stability of the hydrogels, while the pure PVA formulation underperformed in thermal analysis. Additionally, the hydrogels facilitated adequate cell attachment; however, a higher concentration of ZnO NPs led to reduced cell attachment, indicating the cytotoxic behavior of ZnO. The hydrogel incorporating HA and 1 % ZnO demonstrated significant potential as a candidate for wound care applications, indicating the need for further investigation into its in vivo behavior and antimicrobial properties.