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The purpose of this study was to investigate in vitro biocompatibility of a new type of polymer, polyhydroxybutyrate-co-hexanoate (PHBHHx). The hydrophilicity and biocompatibility were studied with two kinds of enzymes, amylase BAN480L and lipase Novozym 388. The degree of hydrophilicity was observed using contact angle measurements. In vitro biocompatibility evaluations were carried out by direct incubation of mouse fibroblast cell line L929 on the polyhydroxyalkanoate (PHA) films. The samples treated with BAN480L showed of the PHA biocompatibility increased while the hydrophilicity decreased. Relative to untreated samples, the number of cells on the Novozym 388 modified PHBHHx significant decrease as the hydrophilicity also decreased. The results indicated that other surface characteristics besides hydrophilicity influence the biocompatibility of PHBHHx films.