In this study, intermolecular crosslinked carboxymethyl cellulose sodium salt (CMC) and porcine Cartilage Acellular Matrix (PCAM) blended hydrogel films for anti-adhesive barriers were prepared by gamma-ray radiation. The effects of the CMC/PCAM concentration and blending ratio on the morphology, gel fraction, gel strength, and degree of swelling were determined. The results indicated that crosslinked CMC/PCAM films show significantly lower the gel-fraction than CMC films. The degree of attachment and proliferation of human vascular endothelial cells on CMC/PCAM films was lower than the CMC films. We show the capacity of the CMC and PCAM to be hydrogel films, and the ability to reduce cell adhesion and proliferation on these films by modification with cell anti-adhesion molecules of PCAM. In conclusion, this study suggests that radiation cross-linked CMC/PCAM hydrogel films endowed with anti-adhesion ligands may allow for improved regulation of cell anti-adhesion behavior for prevent peritoneal adhesions.