Early adhesion of human mesenchymal stem cells on TiO2 surfaces studied by single-cell force spectroscopy measurements

Understanding the interactions involved in the adhesion of living cells on surfaces is essential in the field of tissue engineering and biomaterials. In this study, we investigate the early adhesion of living human mesenchymal stem cells (hMSCs) on flat titanium dioxide (TiO2) and on nanoporous crystallized TiO2 surfaces with the use of atomic force microscopy-based single-cell force spectroscopy measurements. The choice of the substrate surfaces was motivated by the fact that implants widely used in orthopaedic and dental surgery are made in Ti and its alloys. Nanoporous TiO2 surfaces were produced by anodization of Ti surfaces. In a typical force spectroscopy experiment, one living hMSC, immobilized onto a fibronectine-functionalized tipless lever is brought in contact with the surface of interest for 30?s before being detached while recording force-distance curves. Adhesion of hMSCs on nanoporous TiO2 substrates having inner pore diameter of 45?nm was lower by approximately 25% than on TiO2 flat surfaces. Forcedistance curves exhibited also force steps that can be related to the pulling of membrane tethers from the cell membrane. The mean force step was equal to 35?pN for a given speed independently of the substrate surface probed. The number of tethers observed was substrate dependent. Our results suggest that the strength of the initial adhesion between hMSCs and flat or nanoporous TiO2 surfaces is driven by the adsorption of proteins deposited from serum in the culture media. Copyright (C) 2012 John Wiley & Sons, Ltd.