Titanium surfaces were equipped with positively and negatively charged chemical functional groups by plasma polymerization. Their capability to influence the adhesion of human mesenchymal stem cells (hMSCs) and inflammation processes was investigated on titanium substrates, which are representative of real implant surfaces.For these purposes, titanium samples were coated with plasma polymers from allylamine (PPAAm) and acrylic acid (PPAAc). The process development was accompanied by physicochemical surface analysis using XPS, FT-IR and contact angle measurements. Very thin plasma polymer coatings were created, which are resistant to hydrolysis and delamination. Positively charged amino groups improve considerably the initial adhesion and spreading steps of hMSCs. PPAAm and PPAAc surfaces have an effect on the differentiation of hMSCs, e.g., the expression of osteogenic markers in dependence on culturing conditions. Acrylic acid groups appear to stimulate early mRNA differentiation markers (ALP, COL, Runx2) under basal conditions, whereas positively and negatively charged groups both stimulate late differentiation markers, like BSP and OCN, after 3 days of osteogenic stimulation.Long-term intramuscular implantation in rats revealed that PPAAc surfaces caused significantly stronger reactions by macrophages and antigen-presenting cells compared to untreated control (polished titanium) samples, while PPAAm films did not show a negative influence on the inflammatory reaction after implantation.
In dentistry, zirconia has been used since the early 1990s for endodontic posts, more recently for implant abutments and frameworks for fixed dental prostheses. Zirconia is biocompatible and mechanically strong enough to serve as implant material for oral implants. Although several zirconia implant systems are available, currently the scientific and clinical data for zirconia implants are not sufficient to recommend them for routine clinical use. Here the influence of microstructured yttria-stabilized zirconia (YZ) on human primary osteoblast (HOB) behavior was determined. YZ surfaces were treated by sandblasting (YZ-S), acid etching (YZ-SE) and additionally heat treatment (YZ-SEH). Morphological changes of HOB were determined by scanning electron microscopy. Actin cytoskeleton was investigated by laser scanning microscopy and analyzed by novel actin quantification software. Differentiation of HOB was determined by real time RT-PCR. Improved mechanical interlocking of primary HOB into the porous microstructure of the acid etched and additionally heat treated YZ-surfaces correlates with drastically increased osteocalcin (OCN) gene expression. In particular, OCN was considerably elevated in primary HOB after 3 days on YZ-SE (13-fold) as well as YZ-SEH (12-fold) surfaces. Shorter actin filaments without any favored orientation on YZ-SE and YZ-SEH surfaces are associated with higher roughness (Ra) values. Topographically modified yttria-stabilized zirconia is a likely material for dental implants with cell stimulating properties achieving or actually exceeding those of titanium.
Research in plasma medicine includes a major interest in understanding gas plasma-cell interactions. The immediate application of gas plasma in vitro inhibits cell attachment, vitality and cell-cell contacts via the liquid. Interestingly, in our novel experiments described here we found that the liquid-mediated plasma effect is long-lasting after storage up to seven days; i. e. the liquid preserves the characteristics once induced by the argon plasma. Therefore, the complete Dulbecco's Modified Eagle cell culture medium was argon plasma-treated (atmospheric pressure, kINPen09) for 60 s, stored for several days (1, 4 and 7 d) at 37°C and added to a confluent mouse hepatocyte epithelial cell (mHepR1) monolayer. Impaired tight junction architecture as well as shortened microvilli on the cell membrane could be observed, which was accompanied by the loss of cell adhesion capacity. Online-monitoring of vital cells revealed a reduced cell respiration. Our first time-dependent analysis of plasma-treated medium revealed that temperature, hydrogen peroxide production, pH and oxygen content can be excluded as initiators of cell physiological and morphological changes. The here observed persisting biological effects in plasma-treated liquids could open new medical applications in dentistry and orthopaedics.
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