Anodized-hydrothermally treated titanium with a nanotopographic surface structure regulates integrin-α6β4 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-α6β4 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-α6β4 and laminin-5 (α3, β3, 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-α6β4 and laminin-5 (α3, β3, 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-α6β4 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.