Wounds exhibit varied behaviour and types and each type has its own differential healing requirements. This realization has encouraged the development of various wound dressings, each with specific characteristics. In the present study, composite hydrogels composed of PVA/Pullulan/Poly-L-Lysine/Gelatin (P/Pu/L/G) were fabricated using the freeze-thawing method with the aim of achieving enhanced wound healing. The hydrogels were evaluated for their physicochemical and in vitro biological properties. The morphological evaluation using SEM revealed the porous three-dimensional structure exhibited by the P/Pu/L/G hydrogels. The wettability and chemical composition of the hydrogels was elucidated by contact angle and ATR-FTIR analysis respectively, where the contact angle measurements showed an increase in the hydrophilicity of P/Pu hydrogel upon the incorporation of Poly-L-lysine and Gelatin. The results of in vitro hemolysis assay showed better blood compatibility of the fabricated hydrogels while the protein adsorption study revealed a 2.3-fold increase in the protein adsorption on the P/Pu/L/G hydrogels as compared to PVA hydrogels. The composite P/Pu/L/G scaffolds exhibited a 2-fold increase in the cell viability on day 7, when compared to the control; while the results of the scratch assay where complementary to the cell viability assay, which affirmed improved cell migration and proliferation in the presence of P/Pu/L/G hydrogels. The overall results of physicochemical characterization and biological evaluation advocates the potential of the composite hydrogels of P/Pu/L/G for biomedical applications such as wound dressing.