This study was performed to improve the bioactivity of anodized nanotubular TiO₂ layer. The glycerol solution containing 1 wt% NH4F and 20 wt% deionized water was used as an electrolyte. Pulse signals with a potential of 20 V and current density of 20 ㎃/㎠ were applied for 60 minutes. Nanotubular TiO₂ layer was heat-treated at 500℃ for 2 hours and precalcified by soaking in Na₂HPO₄ solution at room temperature for 24 hours and then in saturated Ca(OH)₂ solution for 5 hours. The samples were divided into 4 groups: (1) pure Ti; (2) anodized Ti; (3) anodized and heat-treated; and (4) anodized, heat-treated and precalcified. The 36 implants were placed in both tibiae of 18 rats. After implantation periods of 2 and 4 weeks, the degree of osseointegration of implants were evaluated by micro CT analysis, western blotting and RT-PCR. Micro CT images revealed more dense appearance around screw part of implants in the anodized, heat-treated, and precalcified group than in other groups. Bone mineral density and trabecular density values exhibited coherent results with same oder. Collagen Type I and 2 as well as osteocalcin which are involved in bone remodeling also exhibited the highest expression in the anodized, heat-treated, & precalcified group than in other groups. In conclusion, anodized, heat-treated, and precalcified titanium implants significantly improved osseointegration ability in compared to others.