The optimal temperature for the alkaline treatment and subsequent heat treatment is determined to optimize the nanoporous structures formed on Ti6Al4V titanium alloy plates. Surface characterization of the alkali-heat treated samples was performed by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and X-ray diffraction. The effects of heating temperatures on albumin adhesion, rat bone marrow mesenchymal stem cells (BMMSCs) adhesion, alkaline phosphatase

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... ctivity, osteocalcin production, calcium deposition, and Runx2 mRNA expression were evaluated. The nanotopography, surface chemistry, and surface roughness were unchanged even after heat treatments at 200, 400, and 600°C. Only the amorphous sodium titanate phase changed, increasing with the temperature of the heat treatments, which played a crucial role in promoting superior cell adhesion on the nanoporous surface compared with the sodium hydrogen titanate obtained by a single alkali treatment. The heat treatment at 800°C did not enhance cell attachment on the surface because the nanostructure was dramatically destroyed with the reappearance of Al and V. This study reveals that nanoporous structures with amorphous sodium titanate were fabricated on Ti6Al4V surface through an amended alkali-heat treatment process to improve BMMSCs adhesion.