A hydroxyapatite (HA) film with or without collagen was electrochemically deposited on a bioactivated titanium metal prepared by acid-alkali treatment, so as to improve the biocompatibility of bioactive titanium metals. The cell response of the film was studied with MG63 osteoblasts culture. It was found that the hydroxyapatite formation in the film during the deposition process was inhibited when collagen was added in the electrolyte. More hydroxyapatite with and without collagen could be deposited on the bioactivated titanium than the control titanium metal without treatment, which indicated that the bioactivation process before the electrochemical deposition could accelerate the deposition. The abilities of cell attachment and proliferation were improved by the film especially in the group containing collagen, and the film on the bioactivated metal had higher cell response ability than that on the titanium without treatment. The results indicated that the hydroxyapatite/collagen film could improve the biocompatibility of the bioactive titanium metal surface, and the bioactivation surface modification could further regulate the film and its cell response. It is possible to get a titanium surface with higher bioactivity than the traditional bioactive titanium surface by combining the bioactivation surface modification and electrochemical deposition HA/collagen film.