Biopolymer-based blends can be used within the form of cast film in various proportions with polyethylene oxide (PEO) in order to obtain more durable and biocompatible surfaces. Chitosan and hyaluronic acid (HA) are the most used biocompatible biopolymers for the production of films. On the other hand, as a synthetic polymer, PEO has been mostly used to improve durability of the surfaces by using in various molecular weights. This study was carried out to determine the effect of HA on the chitosan and PEO biopolymer blends different from reported Chitosan/PEO film surfaces in the literature. FTIR studies presented no any polycationic amine groups belong to chitosan through the surface of four films in different compositions after casting. Surface morphology were determined by SEM, EDS and polarized microscopy analyses. Surface morphology was observed as well-oriented spherulitic crystallization by the increasing amount of PEO in the films. Astonishingly, it was proven that HA improved the biocompatibility feature of PEO-rich surfaces which were tested by using healthy mouse fibroblast cell lines, L929 (ATCC CCL-1).