The composition of the dressings is based on polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), and agar. The electron beam irradiation technique has been used to prepare hydrogel wound dressings. The in vitro biocompatibility of the hydrogel was investigated by check samples (hydrocolloid Comfeel), antibacterial test (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia Coli k12), anti fungal test (Candida Albicans) and cytotoxicity test (Fibroblast L929). Results have shown cell attachment characteristics and nontoxicity of all samples. Antibacterial testing also showed that the antibacterial effect of the hydrogel sample to the check sample increased to 30%. Also, investigation of antifungal analysis did not show any trace of fungi growth on the surface of the hydrogel, whereas antifungal effect did not observe on the surface of the check sample. Finally, this hydrogel sample showed a good in vitro biocompatibility.