As reported previously, bioactive titanium is prepared by simple alkali and heat treatment, and can bond to living bone directly. The purpose of this study was to accelerate the bioactivity of bioactive titanium in vivo. In in vitro study, sodium removal by hot water immersion enhanced the apatite-forming ability of bioactive titanium in simulated body fluid dramatically. The specific anatase structure of titania gel was effective for apatite formation in vitro. In the current study, we investigated the in vivo effect of sodium removal on the bone-bonding strength of bioactive titanium. Sodium-free bioactive titanium plates were prepared by immersion in an aqueous solution of 5 M NaOH at 60 degrees C for 24 h, followed by immersion in distilled water at 40 degrees C for 48 h before heating them at 600 degrees C for 1 h. Three kinds of titanium plates were inserted into rabbit tibiae, including untreated cp-Ti, conventional alkali- and heat-treated Ti, and sodium-free alkali- and heat-treated Ti. In vivo bioactive performance was examined mechanically and histologically after 4, 8, 16, and 24 weeks. Sodium removal enhanced the bone-bonding strength of bioactive titanium at 4 and 8 weeks postoperatively; however, its bone-bonding strength was inferior to that of conventional alkali- and heat-treated titanium at 16 and 24 weeks. Histological examinations after the detaching test revealed breakage of the treated layer in the sodium-free alkali- and heat-treated titanium group. In conclusion, sodium removal accelerated the in vivo bioactivity of bioactive titanium and achieved faster bone-bonding because of its anatase surface structure, but the loss of the surface's graded structure due to the complete removal of sodium decreased the adhesive strength of the treated layer to the titanium substrate. Further investigations are required to determine the optimum conditions for preparation of bioactive titanium.