In this research study, novel hydrogel composite films were constructed using different ratios of poly (vinyl alcohol) (PVA)/kappa-carrageenan (KC) (PVA90/KC10%, PVA80/KC20%, PVA70/KC30%, PVA60/KC40%) crosslinked with glutaraldehyde (0.025%) and investigated their physicochemical characteristics such as mechanical, thermal, morphological, swelling behaviour, and cell viability. SEM and FTIR revealed that surface morphology changed to heterogeneous and the presence of molecular interaction among the polymers. PVA90KC10 and PVA60KC40 exhibited smaller and larger pores on surface respectively. The change in the proportion of PVA and KC also triggered the tensile strength (Ts) of the film and the highest Ts observed were 21.60 MPa for PVA60KC40. Moreover, the thermal analysis showed three-phase degradation, and an increase in KC40 concentration results inversely proportional to a decrease in the rate of thermal degradation. Further, swelling and in-vitro biodegradation study confirmed the enhanced perseverance of water uptake for PVA60KC40 (286%) due to pores structure of the hydrogel film and PVA and KC alone degraded faster as compare to other films results suggested higher concentration of PVA90KC10 showed lower degradation rate and highest for PVA60KC40 about 6% and 22% respectively. Further, the cell viability was studied with MTT assay method by using NIH3T3 and HEK-293 cells for biocompatibility study revealed NIH3T3 cells were more biocompatible than HEK-293 and cell viability percent for PVA60KC40 showed the highest cell attachment about 99%. Overall corroborating data obtained from the study attested to the average swelling, appreciable mechanical characters, good interaction between molecules, and cell viability of the constructed PVA/KC hydrogel film, these all characters pave to be used as a potential template for biomedical applications such as tissue engineering and drug delivery.