Abstract The biomechanical stability of mineralized tissues at the interface between implant surface and bone tissue is of critical importance. Anodically oxidized titanium prepared in a chloride solution results in enhanced mineralization of adherent osteoblasts and has antimicrobial activity against oral microorganisms. We evaluated the nanomechanical properties and molecular structures of the in vitro mineralized tissues developing around anodically oxidized titanium surfaces with and without preparation in chloride solution. Anodically oxidized titanium surfaces showed superior osteogenic gene expressions than those of thermally oxidized and bare titanium surfaces. Preparation of anodically oxidized titanium in chloride enhanced the production of mineralized tissue around it. However, the mineralized tissue around anodically oxidized titanium prepared without chloride had increased mineral:matrix and cross-linking ratios, resulting in higher hardness and lower elasticity.