HA/45S5 composite films were deposited by pulsed laser, the crystalline phases, microstructure and bonding configurations of the films were studied by XRD, SEM and FTIR respectively, and the film-to-substrate adhesion was investigated by micro-scratch testing. In addition, the in vitro and in vivo assays were carried out.The results showed that the crystallinity and the adhesive strength of the films increased with the increase of the substrate temperature. The film deposited with the substrate temperature of 200°C is amorphous, while crystalline HA and β-TCP were detected in the film deposited at 600°C. The in vitro test indicates that the amorphous film has faster dissolution rate and reprecipitation rate, which implies that this film has better bioactivity than the crystalline film. However, the in vivo test suggested that the 600°C film was more suitable to serve as clinical application than the 200°C film, because new bone tissue grew better onto the 600°C film surface than onto the 200°C film surface when they were implanted in the rabbit shin bones.