Anodized-hydrothermally treated titanium with a nanotopographic surface structure regulates integrin-α6ß4 and laminin-5 gene expression in adherent murine gingival epithelial cells.

PURPOSE: Peri-implant epithelium associated with the structure of the internal basal lamina is in contact with a transmucosal portion of the endosseous implant surface. This contact is important to protect the many complex factors required for the long-term stability and maintenance of the implant. This study investigated the effect of initial adhesion of gingival epithelial cells to anodized-hydrothermally treated commercially pure titanium with nanotopographic structure (SA-treated c.p.Ti). Changes in cell morphology and gene expression of integrin-α6ß4 and laminin-5 were assessed. METHODS: Murine immortalized gingival epithelial (GE1) cells were cultured for 1-3 days on c.p.Ti, anodic oxide (AO) c.p.Ti, and SA-treated c.p.Ti disks. Cell morphology was analyzed using scanning electron microscopy (SEM). Cell proliferation was analyzed using the WST-1 assay. Integrin-α6ß4 and laminin-5 (α3, ß3, γ2) mRNA levels were measured using real-time quantitative RT-PCR. RESULTS: The GE1 cells appeared flattened with extensions on all disks by SEM analysis. Filopodium-like extensions were bound closely to the nanotopographic structure surface of SA-treated c.p.Ti especially at day 3 of culture. GE1 cell proliferation as well as the expression of integrin-α6ß4 and laminin-5 (α3, ß3, γ2) mRNAs was significantly higher on SA-treated c.p.Ti than on c.p.Ti and AO c.p.Ti disks after 3 days (P<0.05). CONCLUSIONS: Gingival epithelial cells initially attach to a transmucosal portion of SA-treated c.p.Ti implant material and subsequently express the integrin-α6ß4 adhesion molecule and the laminin-5 extracellular matrix molecule. This cell behavior may play a key role in maintaining the peri-implant oral mucosal tissue barrier.

Physicochemical state of the nanotopographic surface of commercially pure titanium following anodization-hydrothermal treatment reveals significantly improved hydrophilicity and surface energy profiles.

A method of coating commercially pure titanium (cpTi) implants with a highly crystalline, thin hydroxyapatite (HA) layer using discharge anodic oxidation followed by hydrothermal treatment (Spark discharged Anodic oxidation treatment ; SA-treated cpTi) has been reported for use in clinical dentistry. We hypothesized that a thin HA layer with high crystallinity and nanostructured anodic titanium oxide film on such SA-treated cpTi implant surfaces might be a crucial function of their surface-specific potential energy. To test this, we analyzed anodic oxide (AO) cpTi and SA-treated cpTi disks by SEM and AFM. Contact angles and surface free energy of each disk surface was measured using FAMAS software. High-magnification SEM and AFM revealed the nanotopographic structure of the anodic titanium oxide film on SA-treated cpTi; however, this was not observed on the AO cpTi surface. The contact angle and surface free energy measurements were also significantly different between AO cpTi and SA-treated cpTi surfaces (Tukey's, P<0.05). These data indicated that the change of physicochemical properties of an anodic titanium oxide film with HA crystals on an SA-treated cpTi surface may play a key role in the phenomenon of osteoconduction during the process of osseointegration.