Abstract The application of pure polyvinyl alcohol (PVA) hydrogel as wound dressing faces many restrictions due to its insufficient elasticity, stiff membrane, and very limited hydrophilicity. These drawbacks can be limited through cross-linking with other natural biopolymers such as gum Arabic (GA). PVA hydrogels blended with six different GA concentrations were prepared. The characterization of these hydrogels was performed by Fourier transform infrared spectrophotometer, X-ray diffraction, and scanning electron microscope. The swelling ratio (% SR) percentage has been calculated. The possible cytotoxicity was determined using a sulforhodamine B assay. Wound healing test was performed on human skin fibroblast cells. Our results indicated that by increasing GA concentration in PVA hydrogel, the % SR increases and the cytotoxicity effect decreases. The results indicate also a significant gradual decrease in the wound area with time as the GA concentration increases in the PVA hydrogel. Therefore, GA improves the biological applications of PVA hydrogel.