In this study the micro-arc oxidized (MAO) and hydrothermally synthesized (MAO-HS) methods were used to modify the TiO2 layer of titanium. The surface characters and the effects on cell behavior, which included cell adhesion kinetics and cytoskeletal organizations were need to evaluate. The surface energy was examined by physicochemical methods. Biological assays were performed with the mouse osteoblastic cell line MC3T3-E1, cells were inoculated on the treated surface of the materials. The surface energy of MAO (50.6 mJ/m2) and MAO-HS (58.5 mJ/m2) was higher than titanium (39.6 mJ/m2 ). A significantly higher cell adhesion rate was observed on the MAO-HS surface than on the MAO and titanium surface. The cytoskeletal organizations were analyzed by actin and vinculin staining on all the samples after being cultured 2 hours. Modification of TiO2 layer on titanium surface has an influence on the surface characters and the cell behavior on the materials.