Purpose: The objectives of this study were to evaluate bioactivity of a titanium membrane with anodization, cyclic precalcification, and heat (APH) treatment (APHTM), and to compare APHTM and nontreated titanium membrane (NTTM) in guided bone regeneration using histologic analysis and microcomputed tomography (micro-CT). Materials and Methods: APHTM samples were prepared and immersed in simulated body fluid for 2 days, then observed using field-emission scanning electron microscopy, followed by an analysis of calcium and phosphate precipitation using an energy dispersive x-ray spectroscopy. For the in vivo experiment, critical-size defects were created in rat calvaria (diameter, 8 mm) and treated with either APHTM or NTTM (n = 14 each). Biopsies were performed at 2 and 4 weeks for histologic analysis (n = 3 per group each time). Fluorochrome bone markers were injected in three rats in each group at 3 (alizarin red) and 5 weeks (calcein green), followed by histologic examination at 7 weeks. Micro-CT was performed at 8 weeks (n = 5 per group). Results: APHTM exhibited high bioactivity, characterized by dense nano-sized flakelike crystals throughout the membrane and an increase in the calcium-phosphate concentrations after 2-day immersion in simulated body fluid. At 2 and 4 weeks, APHTM samples showed an intimate bone formation onto the membrane, whereas NTTM samples demonstrated interposition of connective tissue between the membrane and newly formed bone. The same pattern was found in the fluorescent study. The micro-CT analysis revealed significantly lower bone volume but higher bone mineral density in the APHTM samples than in the NTTM samples (P .05). Conclusion: The results suggest that APH treatment on titanium membrane promotes intimate bone formation toward the membrane, thus increasing structural durability for bone regeneration. Further research is warranted to confirm the results found in these in vitro and in vivo experiments. Keywords: calcium-phosphate coating, fluorochrome labeling, guided bone regeneration, rat calvarial model, titanium membrane