Responses of human gingival fibroblasts to superhydrophilic hydrogenated titanium dioxide nanotubes

Soft tissue integration is critical for the long-term retention of dental implants. The surface properties including topography and wettability can impact soft tissue sealing. In our work, a thermal hydrogenation technique was applied to modify anodized titanium dioxide nanotubes (TNTs). However, the effects of the hydrogenated surface on soft-tissue cells remain unclear. The aim of the present study was to investigate the bioactivities of human gingival fibroblasts (HGFs) on structured surfaces, which determine the early formation of soft tissue sealing. Three groups were examined: commercially pure titanium (Ti), anodized TNTs (air-TNTs) and hydrogenated TNTs (H₂-TNTs). Scanning electron microscopy showed the nanotubular topography on the titanium surfaces after anodization. Then, hydrogenation ensured that the H₂-TNTs were superhydrophilic with a contact angle of 3.5 ± 0.8°. In vitro studies such as cell adhesion assays, cell morphology, immunocytochemistry, wound healing assays, real-time PCR and enzyme-linked immunosorbent assays displayed enhanced adhesion, migration, relative gene expression levels, and extracellular matrix synthesis of the HGFs on H₂-TNTs. Interestingly, focal adhesion kinase activation and integrin-mediated adhesion seemed to be induced by the H₂-TNT surface. Our results revealed that a superhydrophilic nanostructure modified by anodization and hydrogenation can improve the bioactivity of HGFs and connective tissue regeneration, which will further promote and expand the application of titanium dioxide nanotubes in dental implants.