Due to the rising demand for zirconia (Zr) based implant systems, it is important to understand the impact of Zr and titanium (Ti) implants and particularly their topography on soft tissue healing. As human gingival fibroblasts (hGFs) are the predominant cells in peri-implant soft tissue, we focused on examining the effect of implant material and surface roughness on hGFs’ initial attachment, growth and the expression of proteins involved in the focal adhesion. hGFs isolated from eight healthy donors were cultured on the following surfaces: smooth titanium machined surface (TiM), smooth zirconia machined surface (ZrM), moderately rough titanium surface (SLA), or moderately rough zirconia surface (ZLA) for up to 14 days. The initial attachment of hGFs was evaluated by scanning electron microscopy. Cell proliferation/viability was assessed by cell counting kit 8. Focal adhesion and cytoskeleton were visualized by a focal adhesion staining kit. The gene expression of focal adhesion kinase (FAK), α-smooth muscle actin (α-SMA), and integrin subunits ITG-β1, ITG-β4, ITG-α4, ITG-α5, ITG-α6, was evaluated by qPCR. Cell proliferation/viability was slightly decreased by moderately rough surfaces, whereas no effect of surface material was observed. Cell morphology was strikingly different between differently treated surfaces: on machined surfaces, cells had elongated morphology and were attached along the grooves, whereas on moderately rough surfaces, cells were randomly attached. Surface roughness had a more pronounced effect on the gene expression compared to the surface material. The expression of FAK, α-SMA, ITG-β4, ITG-α5, and ITG-α6 was enhanced by moderately rough surfaces compared to smooth surfaces. Within the limitations of this in vitro study, it can be concluded that the behavior of primary hGFs is primarily affected by surface structure, whereas no apparent advantage of Zr over Ti could be observed.

Orthodontic clear aligner treatment is gaining tremendous popularity. The world market leader is Align Technology^® and its product Invisalign^®. Although numerous patients are treated with Invisalign^® aligners, only little is known about the cellular effects of aligner material on oral epithelial cells. In the present study, we aimed to investigate the effects of SmartTrack^® clear aligner material on directly cultured primary human oral keratinocytes (HOKs). Cell morphology and behavior were investigated by scanning electron microscopy and bright field microscopy. Aligner effects on viability were detected by cell-counting-kit (CCK)-8 and live/dead staining. Gene expression of several inflammatory and barrier proteins was assessed by qPCR. Cells cultured on tissue culture plastic served as control. Cell proliferation/viability was significantly lower in cells cultured on aligner material (p < 0.05) in comparison to control. Live/dead staining did not reveal an increase in the number of dead cells on aligner surfaces. After two and seven days of incubation, interleukin (IL)-6 expression decreased, and IL-8 expression increased in HOKs cultured on aligner surfaces. The expression of intercellular adhesion molecule 1 (ICAM-1) significantly decreased after seven days. Gene expression of epithelial barrier markers showed that integrin (ITG)-α6 significantly decreased after two and seven days. A significant decrease in ITG-β4 and E-cadherin expression levels compared to control could only be seen after seven days. We did not find any cytotoxic effect, but alterations in the cell’s barrier functions and inflammatory reaction were obvious. Clinical studies are required to give further insights into clinical reactions on the underlying aligner material of this quickly expanding orthodontic appliance.

The aim of this study was to evaluate the impact of a novel multi-phosphonate (MP) coating strategy of dental implant surfaces on the expression of osteogenesis-related factors in vitro. MG-63 human osteoblast-like cells, bone marrow mesenchymal stem cells (BM-MSCs), and human periodontal ligament stem cells (hPDLSCs) were cultured separately on titanium disks with and without MP coating. Cell attachment was visualized by focal adhesion and actin cytoskeleton staining. The proliferation and gene expression of the markers related to osteogenesis and bone turnover were measured after 48 and 120 h of cell culture. Actin cytoskeleton assembly and focal adhesion were similar between test surfaces within each cell type but differed from those on tissue culture plastic (TCP). The proliferation of MG-63 cells and PDLSCs was comparable on all surfaces, while BM-MSCs showed an increase on tissue culture plastic (TCP) versus titanium. The gene expression of osteoprotegerin and receptor activator of nuclear factor-kappa B ligand was higher in MG-63 cells grown on MP-coated surfaces. At the same time, osteocalcin was decreased compared to the other surfaces. Collagen type I gene expression after 120 h was significantly lower in hPDLSCs cultivated on MP-coated surfaces. Within the limitations of this study, MP coating on titanium surfaces might have a slight beneficial effect on bone turnover in vitro.