SURFACE MICROGROOVES OF THIRTY MICROMETERS IN WIDTH ON TITANIUM SUBSTRATA ENHANCE PROLIFERATION AND ALTER GENE EXPRESSION OF CULTURED HUMAN GINGIVAL FIBROBLASTS

  • Lee, Suk-Won (Department of Biomaterials & Prosthodontics, Dental Hospital, Kyunghee University, East-West Neo Medical Center) ;
  • Kim, Su-Yeon (Research Institute of Medical Science, St. Vincent's Hospital) ;
  • Lee, Keun-Woo (Department of Prosthodontics, Collage of Dentistry, Yonsei University)
  • Published : 2007.12.31

Abstract

Statement of problem. Surface microgrooves on Ti substrata have been shown to alter the expression of genes responsible for various biological activities of cultured fibroblasts. However, their effect on enhancing cell proliferation is not yet clear. Purpose. The purpose of this study was to determine the dimension of surface microgrooves on Ti substrata that enhances proliferation and alters gene expression of cultured human gingival fibroblasts. Material and methods. Commercially pure Ti discs with surface microgrooves of monotonous $3.5{\mu}m$ in depth and respective 15 and $30{\mu}m$ in width were fabricated using photolithography and used as the culture substrata in the two experimental groups in this study (TiD15 and TiD30), whereas the smooth Ti was used as the control substrata (smooth Ti group). Human gingival fibroblasts were cultured on the three groups of titanium substrata and the proliferation, DNA synthesis, and gene expression of theses cells were analyzed and compared between all groups using XTT assay, BrdU assay, and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. Results. From the XTT assay at 48 h incubation, the proliferation of human gingival fibroblasts in TiD30 was significantly enhanced compared to that in smooth Ti and TiD15. The results from the BrdU assay showed that, at 24 h incubation, the DNA synthesis was significantly enhanced in TiD30 compared to that in smooth Ti. In RT-PCR, increase in the expression of PCR transcripts of fibronectin, CDK6, $p21^{cip1}$ genes was noted at 48h incubation. Conclusion. Surface microgrooves $30{\mu}m$ in width and $3.5{\mu}m$ in depth on Ti substrata enhance proliferation and alter gene expression of cultured human gingival fibroblasts.

Keywords

References

  1. den Braber ET, de Ruijter JE, Ginsel LA, von Recum AF, Jansen JA. Quantitative analysis of fibroblast morphology on micro grooved surfaces with various groove and ridge dimensions. Biomater 1996;17:2037-2044 https://doi.org/10.1016/0142-9612(96)00032-4
  2. Walboomers XF, Ginsel LA, Jansen JA. Early spreading events of fibroblasts on microgrooved substrates. J Biomed Mater Res 2000;51:529-534 https://doi.org/10.1002/1097-4636(20000905)51:3<529::AID-JBM30>3.0.CO;2-R
  3. Chou L, Firth JD, Vitto VJ, Brunette DM. Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts. J Cell Sci 1995;108:1563-1573
  4. Dalby MJ, Riehl, MO, Yarwood SJ, Wilkinson CD Curtis AS. Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography. Exp Cell Res 2003;284:274-282
  5. den Braber ET, de Ruijter JE, Smits HT, Ginsel LA, von Recum AF, Jansen JA. Quantitative analysis of cell proliferation and orientation on substrata with uniform parallel surface micro-grooves. Biomater 1996;17:1093-1099 https://doi.org/10.1016/0142-9612(96)85910-2
  6. Walboomers XF, Monaghan W, Curtis AS, Jansen JA. Attachment of fibroblasts on smooth and microgrooved polystyrene. J Biomed Mater Res 1999;46:212-220 https://doi.org/10.1002/(SICI)1097-4636(199908)46:2<212::AID-JBM10>3.0.CO;2-Y
  7. Walboomers XF, Croes HJ, Ginsel LA, Jansen JA. Contact guidance of rat fibroblasts on various implant materials. J Biomed Mater Res 1999;47:204-212 https://doi.org/10.1002/(SICI)1097-4636(199911)47:2<204::AID-JBM10>3.0.CO;2-H
  8. Zhang C, Meng XF, Zhu ZH, Yang X, Deng AG. Role of connective tissue growth factor in plasminogen activator inhibitor-1 and fibronectin expression induced by transforming growth factor ${\beta}$1 in renal tubular cells. Chinese Med J 2004;117:990-996
  9. Moran C], Friel AM, Smith 1], Cairns M, Morrison JJ. Expression and modulation of Rho kinase in human pregnant myometrium. Molecular Human Reproduction Mol Hum Reprod. 2002;8:196-200 https://doi.org/10.1093/molehr/8.2.196
  10. Boumediene K, Takigawa M, Pujol JP. Cell densitydependent proliferative effects of transforming growth factor (TGF)-beta 1, beta 2, and beta 3 in human chondrosarcoma cells HC5-2/8 are associated with changes in the expression of TGF-beta receptor type I. Cancer Invest 2001;19:475-486 https://doi.org/10.1081/CNV-100103846
  11. Takayama S, Yoshida J, Hirano H, Okada H, Murakami S. Effects of basic fibroblast growth factor on human gingival epithelial cells. J PeriodontoI 2002;73:1467-1473 https://doi.org/10.1902/jop.2002.73.12.1467
  12. Wong H, Riabowol K. Differential CDK-inhibitor gene expression in aging human diploid fibroblasts. Exp GerontoI 1996;3:311-325
  13. Kaiser JP, Reinmann A, Bruinink, A. The effect of topographic characteristics on cell migration velocity. Biomater 2006;27:5230-5241 https://doi.org/10.1016/j.biomaterials.2006.06.002
  14. Lee JW, Juliano R. Mitogenic signal transduction by integrin- and growth factor receptor-mediated pathways. Mol Cells 2004;17:188-202
  15. Li S, Lao J, Chen BP, U YS, Zhao Y, Chu J, Chen KD, Tsou TC, Peck K, Chien S. Genomic analysis of smooth muscle cells in 3-dimensional collagen matrix. FASEB J 2003;17:97-99 https://doi.org/10.1096/fj.02-0256fje