Effect of Titanium Surface Calcium and Magnesium on Adhesive Activity of Epithelial-Like Cells and Fibroblasts

Abstract

In the present study, we investigated the hydrothermal treatment of titanium with divalent cation solutions and its effect in promoting the adhesion of gingival epithelial cells and fibroblasts in vitro. Gingival keratinocyte-like Sa3 cells or fibroblastic NIH3T3 cells were cultured for 1 h on experimental titanium plates hydrothermally-treated with CaCl2 (Ca) or MgCl2 (Mg) solution, or distilled water (DW). The number and adhesive strengths of attached cells on the substrata were then analyzed. The number of Sa3 cells adhering to the Ca- and Mg-treated plates was significantly larger than in the DW group, but the strength of this adhesion did not differ significantly between groups. In contrast, NIH3T3 cell adhesion number and strength were increased in both the Ca and Mg groups compared to the DW group. Fluorescent microscopic observation indicated that, in all groups, Sa3 had identical expression levels of integrin β4 and development of actin filaments, whereas NIH3T3 cells in the Ca and Mg groups displayed much stronger punctate cytoplasmic signals for vinculin and more bundle-shaped actin filaments than cells in the DW group. As a result, it was indicated that the hydrothermal treatment of titanium with Ca or Mg solution improved the integration of soft tissue cells with the substrata, which may facilitate the development of a soft tissue barrier around the implant.

Effects of CaCl2 hydrothermal treatment of titanium implant surfaces on early epithelial sealing

“…5). These results are consistent with previous studies using an early-stage epithelial cell line [12].…”

“…Thus, an apatite layer may not develop. In fact, we did not detect any layers between the cells or tissues and the titanium in the present and previous studies [12,13].…”

“…The hydroxyl groups of the Ti surface have a significant effect on enhancement of protein adsorption [22]. Another study has reported that, regardless of the presence of cations, hydrothermal treatment enriches surface hydroxyl groups [12]. In addition, the presence of CaTiO 3 has been reported on the surface of titanium treated hydrothermally with a CaCl 2 solution [23].…”

“…The three experimental groups included: (1) control (Cont) group specimens, which received no treatment; (2) distilled water (DW)-hydrothermally treated (HT) group specimens, which were hydrothermally treated in DW at 200 • C for 24 h; (3) CaCl 2 -HT specimens (Ca-HT group) that were treated at 200 • C for 24 h with a 10 mmol/L aqueous solution of CaCl 2 as reported previously ( Fig. 1A and B) [10,12,13]. After treatment, all specimens were washed with DW and stored in a vacuum to prevent surface contamination.…”

“…OEC adhesion assays were conducted according to previously published methods [8,12]. Non-adherent or weakly attached cells were removed by shaking (3× 5 min at 75 rpm) using a rotary shaker (NX-20, Nissin, Tokyo, Japan).…”