With good biocompatibility, collagen is often used in cartilage tissue engineering. Collagen/alginate composite was hoped to improve the poor mechanical property of pure collagen but the biocompatibity was decreased. In this study, hydroxyapatite (HA) particles were used to get collagen/alginate/HA (CAHA) composite film to enhance the bioactivity properties. The bioactivity of the composite was investigated by in vitro co-culture with chondrocytes. During the 6-day cell culture in vitro, the composite showed a significant improvement in promoting proliferation and maintaining morphology/phenotype of the chondrocytes over collagen/alginate composite by MTT, SEM, fluorescent and immunohistochemical assays. Cytocompatibility and cytoviablility of CAHA even come up to that of collagen film alone. The results indicated that the composite film may provide an appropriate environment for the proliferation and maintaining the morphology and phenotype of chondrocytes and have a potential clinical application in the cartilage tissue engineering field.