Improving endothelial cell adhesion and proliferation on titanium by sol-gel derived oxide coating.

In-stent restenosis becomes increasingly prevalent as a difficult-to-treat disease. An alternative therapeutic strategy is enhancing endothelialization on metallic stent surfaces. This study attempted to modify surface chemistry and topography of commercial pure titanium (cp-Ti) by different sol-gel derived oxide coatings (TiO(2), SiO(2), SiO(2)/TiO(2), and Nb(2)O(5)) to improve endothelialization. The physiochemical properties of the modified surfaces were characterized by ellipsometry, atomic force microscope, and sessile-drop method. The cell adhesion/proliferation quantity, cell adhesion morphology, and focal adhesion protein expression were evaluated with human pulmonary microvascular endothelial cell line. The thickness of oxide coatings approximates to 100 nm; significantly rougher nanoporous structure was found in the TiO(2) and Nb(2)O(5) coatings than that of cp-Ti. SiO(2) coating possesses the highest surface energy (75.1 mJ/m(2)) and the lowest was for cp-Ti (45.7 mJ/m(2)). TiO(2) coating showed significantly higher endothelial cell adhesion rate than others; TiO(2), Nb(2)O(5), and TiO(2)/SiO(2) coatings exhibited higher endothelial proliferation in 3-day assays than noncoated Ti. In hemocompatible test, they also showed good hemocompatibility. These results offer the insight into that certain oxide coatings on titanium could significantly improve endothelial cell adhesion and proliferation especially in early period, which will favor reaching the endothelialization rapidly and suitable as matrix for "endothelial seeding" stent.

Osteoblast responses to different oxide coatings produced by the sol-gel process on titanium substrates.

In order to improve the osseointegration of endosseous implants made from titanium, the structure and composition of the surface were modified. Mirror-polished commercially pure (cp) titanium substrates were coated by the sol-gel process with different oxides: TiO(2), SiO(2), Nb(2)O(5) and SiO(2)-TiO(2). The coatings were physically and biologically characterized. Infrared spectroscopy confirmed the absence of organic residues. Ellipsometry determined the thickness of layers to be approximately 100nm. High resolution scanning electron microscopy (SEM) and atomice force microscopy revealed a nanoporous structure in the TiO(2) and Nb(2)O(5) layers, whereas the SiO(2) and SiO(2)-TiO(2) layers appeared almost smooth. The R(a) values, as determined by white-light interferometry, ranged from 20 to 50nm. The surface energy determined by the sessile-drop contact angle method revealed the highest polar component for SiO(2) (30.7mJm(-2)) and the lowest for cp-Ti and 316L stainless steel (6.7mJm(-2)). Cytocompatibility of the oxide layers was investigated with MC3T3-E1 osteoblasts in vitro (proliferation, vitality, morphology and cytochemical/immunolabelling of actin and vinculin). Higher cell proliferation rates were found in SiO(2)-TiO(2) and TiO(2), and lower in Nb(2)O(5) and SiO(2); whereas the vitality rates increased for cp-Ti and Nb(2)O(5). Cytochemical assays showed that all substrates induced a normal cytoskeleton and well-developed focal adhesion contacts. SEM revealed good cell attachment for all coating layers. In conclusion, the sol-gel-derived oxide layers were thin, pure and nanostructured; consequent different osteoblast responses to those coatings are explained by the mutual action and coadjustment of different interrelated surface parameters.

Cell type-specific aspects in biocompatibility testing: the intercellular contact in vitro as an indicator for endothelial cell compatibility.

Endothelial cells cover the inner surface of blood vessels and form the interface between the blood and the tissues. Endothelial cells are involved in regulating barrier function, which is maintained by the interendothelial cell contacts. These interendothelial cell contacts are established by the interaction of different molecules. The maintenance of the barrier requires an appropriate signalling between these molecules. Thus, a number of different signalling pathways are integrated within interendothelial contacts. Since endothelial cells are important in tissue-implant interactions (especially for stent materials) this study examines the expression pattern of different interendothelial contact molecules to determine the usefulness in the analysis of biocompatibility in vitro. The effects of different pro-inflammatory and toxic stimuli and contact of human microvascular endothelial cells to metallic surfaces were examined for their impact on the pattern of interendothelial contact molecules. Striking modifications in the arrangement of these molecules were induced and the mode of modification was dependent on the tested compound. Thus, examining the pattern of expression of specific interendothelial contact molecules in vitro may be useful for testing the endothelial cell compatibility of biomaterials and their corrosion products.

Growth behavior, matrix production, and gene expression of human osteoblasts in defined cylindrical titanium channels.

The purpose of the current study was to investigate the effect of different diameters of cylindrical titanium channels on human osteoblasts. Titanium samples having continuous drill channels with diameters of 300, 400, 500, 600, and 1000 microm were put into osteoblast cell cultures that were isolated from 12 adult human trauma patients. Cell migration into the drill channels was investigated by transmitted-light microscopy. The DNA content in the drill channels was measured photometrically, collagen type I production was analyzed by enzyme-linked immunosorbent assay (ELISA) and osteocalcin gene expression by reverse transcriptase-polymerase chain reaction (RT-PCR). Formation of mineralized tissue was assessed by microradiographs of histological sections. Within 20 days, cells grew an average of 838 microm (+/-128 microm) into the drill channels with a diameter of 600 microm and were significantly faster (p < 0.05) than in all other channels. Cells produced significantly more osteocalcin messenger RNA (mRNA) in 600-microm channels (p < 0.05) than they did in 1000-microm channels and demonstrated the highest osteogenic differentiation. The channel diameter did not influence collagen type I production. The highest cell density was found in 300-microm channels (p < 0.05). The DNA content of the channels linearly decreased with increasing channel diameters. After 40 days of culture, the proportion of mineralized tissue at the mouth section amounted to 6% in 300-microm channels and to 9-11% in 400-600-microm channels. In 1000-microm channels, only traces of mineralization were detected. Our data suggest that the diameter of cylindrical titanium channels has a significant effect on migration, gene expression, and mineralization of human osteoblasts.