Regulation of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases by basic fibroblast growth factor and dexamethasone in periodontal ligament cells

J Periodontal Res. 2009 Dec;44(6):794-802. doi: 10.1111/j.1600-0765.2008.01192.x. Epub 2009 Jul 2.

Abstract

Background and objectives: In this study, we investigated the effect of basic fibroblast growth factor (bFGF) and dexamethasone (Dex) on mRNA expressions of collagen (COL) type I, III and X, matrix metalloproteinases (MMP)-1, -2, -3 and -9 and tissue inhibitors of metalloproteinases (TIMP)-1 and -2, and also on mineralization and morphology of periodontal ligament (PDL) cells.

Material and methods: Periodontal ligament cells were obtained from premolar teeth extracted for orthodontic reasons. Periodontal ligament cells were cultured with Dulbecco's modified Eagle's medium containing: (1) 5% fetal bovine serum (FBS); (2) 5% FBS + ascorbic acid (AA, 50 microg/mL); (3) 5% FBS + Dex (10(-7) m) + AA; (4) 5% FBS + bFGF (10 ng/mL) + AA; or (5) 5% FBS + Dex (10(-7) m) + bFGF + AA. Cells within each group were evaluated for gene expression profile using semi-quantitative reverse transcriptase-polymerase chain reaction for COL I, III and X, MMP-1, -2, -3 and -9 and TIMP-1 and -2 on days 14 and 21 and for biomineralization by von Kossa stain in vitro on day 21. Images of PDL cells were examined using a phase contrast microscope.

Results: Basic fibroblast growth factor stimulated MMP-1, MMP-3 and MMP-9 mRNA expressions and inhibited TIMP-2 mRNA expression. Treatment of cells with Dex + bFGF led to downregulation of MMP-1, MMP-3 and MMP-9 transcripts. Whilst AA alone and Dex alone induced biomineralization of PDL cells, bFGF blocked the mineralization activity of the cells. In the Dex + bFGF group, more mineral nodules were noted when compared to AA alone and Dex alone groups.

Conclusion: The addition of Dex to culture reversed bFGF-mediated inhibition of mineralization. Use of combined bFGF and Dex to regulate PDL cell function may be a good therapeutic option to obtain periodontal regeneration.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcification, Physiologic / drug effects
  • Cell Shape / drug effects
  • Cells, Cultured
  • Collagen Type I / drug effects
  • Collagen Type III / drug effects
  • Collagen Type X / drug effects
  • DNA / drug effects
  • Dexamethasone / pharmacology*
  • Down-Regulation
  • Fibroblast Growth Factor 2 / pharmacology*
  • Gene Expression Profiling
  • Glucocorticoids / pharmacology*
  • Humans
  • Matrix Metalloproteinase 1 / drug effects
  • Matrix Metalloproteinase 2 / drug effects
  • Matrix Metalloproteinase 3 / drug effects
  • Matrix Metalloproteinase 9 / drug effects
  • Matrix Metalloproteinases / drug effects*
  • Periodontal Ligament / cytology
  • Periodontal Ligament / drug effects*
  • RNA, Messenger / drug effects
  • Tissue Inhibitor of Metalloproteinase-1 / drug effects
  • Tissue Inhibitor of Metalloproteinase-2 / antagonists & inhibitors
  • Tissue Inhibitor of Metalloproteinase-2 / drug effects
  • Tissue Inhibitor of Metalloproteinases / drug effects*

Substances

  • Collagen Type I
  • Collagen Type III
  • Collagen Type X
  • Glucocorticoids
  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinase-1
  • Tissue Inhibitor of Metalloproteinases
  • Fibroblast Growth Factor 2
  • Tissue Inhibitor of Metalloproteinase-2
  • Dexamethasone
  • DNA
  • Matrix Metalloproteinases
  • MMP3 protein, human
  • Matrix Metalloproteinase 3
  • MMP2 protein, human
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9
  • MMP1 protein, human
  • Matrix Metalloproteinase 1