Micropatterned grooves and acid-etching on titanium substrata alterviability and gene expression of adhered human gingival fibroblasts: A pilot study

Abstract

Statement of problem. Prior to determining an optimal width of micropatterned grooves provided on titanium substrata, we have done a pilot study using surface topographies in combined microm and submicrom levels.

Purpose. The purpose of this study was twofold 1) to assess the proliferation and 2) to analyze the expression of genes encoding the intracellular signaling proteins involved in cell-substratum adhesions and adhesion-dependent G1 phase cell cycle progression of human gingival fibroblasts plated on smooth and microgrooved/acid-etched titanium substrata.

Material and methods. Three groups of titanium discs as NE0 (smooth Ti substrata), E15 (Ti substrata with microgrooves of 15{mu}m of spacing and 3.5{mu}m in depth and with further acidetching), and E30 (Ti substrata with microgrooves of 30{mu}m spacing and 3.5{mu}m in depth and with further acid-etching) served as the human gingival fibroblasts’ substrata. Viability and proliferation of fibroblasts were determined using an XTT assay. Gene expressions of fibronectin, {alpha}5 integrin, CDK4, and p27^{kip} were analyzed in RT-PCR. Cell-substratum interactions were analyzed in SEM.

Results. From the XTT assay at 24 h incubation, the mean optical density (OD) value of E15 was significantly greater than the values of E30 and NE0. At 48 and 96 h however, the mean OD values of E30 were significantly greater than the values of E15 and NE0. No differences in the expression of PCR transcripts at 96 h incubations were noted between groups, whereas at 48 h, an unexpected increase in the expression of all the transcripts were noted in E15 compared with other two groups. Fibroblasts were observed to orient and adhere inside the microgrooves.

Conclusion. Micropatterned grooves and acid-etching on Ti substrata alter viability and gene expression of adhered human gingival fibroblasts.