Vitronectin adsorption on surfaces visualized by tapping mode atomic force microscopy.
Vitronectin (Vn) is an abundant glycoprotein present in plasma and the extracellular matrix of most tissue and is an important medium required for bone cells to attach and spread on biomaterial surfaces. Hence, Vn adsorption is an initial and key step in implantation. In this study, adsorption of Vn molecules on mica and oxidized titanium substrates in Milli-Q water (pH 5.6) and in simulated body fluid (SBF, pH 7.4) has been investigated using tapping mode atomic force microscopy. Conformation of the adsorbed proteins was determined from the images. The effect of adsorption time was investigated on mica surfaces. The protein first attaches to the surface, then immediately spreads out with different lateral dimensions of adsorbed protein and becomes aggregated. After 15 min of adsorption, globular Vn molecules form clusters of aggregates in extended chains on the mica surface. Globular Vn molecules appear flatter (i.e., oblate ellipsoids) and coalesce on the ridges of the uneven oxidized titanium surface. From cross-sectional analyses of the Vn images, it was found that the contact areas of Vn molecules associated with Ti surfaces are larger than those with mica surfaces. This suggests that the different surface properties of substrates contribute to this different conformation of adsorbed Vn molecules. The larger contact areas of Vn associated with Ti substrates indicates higher affinity for the biomaterial oxidized titanium surface than for mica. In SBF, Vn interacted weakly with the mica surfaces and Vn molecules were easily desorbed during the imaging process. On oxidized titanium substrates, after 3 h of incubation time in Vn-SBF solution, images can be obtained showing adsorbed Vn aggregates on the oxidized titanium surface. In this case, the protein colloids are strongly adherent, with increased lateral dimensions compared with the adsorption on mica.
Zhang
Hailong
H
Ian Wark Research Institute, University of South Australia, Mawson Lakes, South Australia 5095, Australia.
Bremmell
Kristen
K
Kumar
Sunil
S
Smart
Roger St C
RS
D000327:Adsorption; D000538:Aluminum Silicates; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism*; D001826:Body Fluids; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D010949:Plasma; D011485:Protein Binding; D011487:Protein Conformation; D013499:Surface Properties; D014025:Titanium; D019096:Vitronectin / Q000737:chemistry / Q000378:metabolism*
D000538:Aluminum Silicates; D001672:Biocompatible Materials; D019096:Vitronectin; C009495:titanium dioxide; D014025:Titanium; C011934:mica