Surface analyses of micro-arc oxidized and hydrothermally treated titanium and effect on osteoblast behavior

Osteoblast adhesion on the implant material surface is essential for the success of any implant in which osteointegration is required. Surface properties of implant material have a critical role in the cell adhesion progress. Titanium and its alloys are widespread and increasingly used as implant material in dentistry and orthopedics because of their excellent biocompatibility, which is attributed to a passive layer of TiO₂ on the surface. In this study, the micro-arc oxidizing (MAO) and hydrothermally synthesizing (HS) methods were used to modify the TiO₂ layer on the titanium surface. The surface microstructure was observed by scanning electron microscopy. The surface energy was assessed. The mouse osteoblastic cell line (MC3T3-E1) was seeded on the treated surfaces to evaluate their effect on cell behavior. This included cell adhesion kinetics, cell proliferation, cell morphology, and cytoskeletal organization. The surface structure of MAO samples exhibited micropores with a diameter of 1-3 μm, whereas the MAO-HS-treated samples showed additional multiple crystalline microparticles on the microporous surface. The surface energy of MAO and MAO-HS was higher than that of titanium. The cell adhesion rate was higher on the MAO-HS surface than on the MAO and titanium surface, but without any significant difference between them. After 3 days of culture, cells proliferated significantly more on the MAO and titanium surface than on the MAO-HS surface. The cytoskeletal organization was analyzed by actin and vinculin staining on all the samples. We conclude that the MAO and MAO-HS methods change the surface energy of TiO₂ layer on the titanium surface. This may have an influence on the initial cell attachment. Other surface characteristics may be involved in the cell proliferation, which is different from cell attachment on the sample surface. A longer-duration cell experiment should be conducted to see the effect on cell differentiation. Future in vivo evaluation may give further evidence to optimize the surface character of this kind of implant material.