In order to improve hydrophilicity and biocompatibility of chitosan, hyaluronic acid was immobilized onto the surface of chitosan film. The structure of films was characterized by Fourier transformed infrared spectroscopy with attenuated total reflectance (ATR-FTIR), x-ray photoelectron spectroscopy (XPS) and zeta potential. Results confirmed that hyaluronic acid was successfully immobilized on chitosan film. Transparency, water absorption percentage and contact angle of films were characterized. Results showed that there was no significant variation in transparency (p < 0.05) before and after immobilization, the maximum was up to 99% which was enough for corneal regeneration in clinical applications. After the immobilization, the time-dependent contact angle declined sharply (from 91.8 degrees to 67.7 degrees at 100 s). The hydrophilicity was significantly improved. The methylthiazol tetrazolium (MTT) (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was used to assess cell viability and proliferation. Results showed that human cornea epithelial cells (HCEC) grew better on hyaluronic acid immobilized chitosan films than on chitosan films. The hyaluronic acid immobilized chitosan film could be a promising candidate material for corneal regeneration.