Enhanced initial adhesion of osteoblast-like cells on an anatase-structured titania surface formed by H₂O₂/HCl solution and heat treatment

Abstract 

Objectives

This study aims to investigate the effects of anatase-structured titania surface on the response of osteoblast-like cells.

Methods

Three kinds of titanium (Ti) disks were prepared: the polished Ti (PT), the roughened Ti (RT) formed by sandblasting and acid etching, and the anatase-coated roughened Ti (ART) formed on the RT via H₂O₂/HCl solution treatment followed by heating at 400°C. The surface topography of Ti disks was observed under a scanning electron microscope. MC3T3-E1 cells were cultured on the Ti disks. Cell morphology was observed by fluorescence microscopy. Cell adhesion and proliferation were assessed by MTT testing. To evaluate the cell differentiation on RT and ART, the expressions of core-binding factor α, osterix, bone sialoprotein, collagen type I????, osteocalcin and alkaline phosphatase-2 were measured by real-time reverse transcriptase polymerase chain reaction.

Results

The three kinds of Ti disks had distinct surface topographic characteristics. The cells on the ART more rapidly displayed well-defined stress fibers and numerous processes and pseudopodi than the cells on the PT and RT. At 1 and 3h after culture, there were more cells adhering to the ART than to PT and RT. Cell proliferation did not differ between groups after 1–5 days of culture. In addition, ART generally enhanced the osteogenic gene expression in a time-dependent manner compared with RT.

Significance

Anatase-structured titania promotes initial adhesion and spreading of osteoblast-like cells, but does not influence cell proliferation. Anatase could also enhance cell differentiation by enhancing osteogenic gene expression.

Keywords: Anatase, Surface topography, Osteoblast, Cell adhesion, Osteogenic genes