Temporomandibular joint synovial fibroblasts mediate serine proteinase dependent Type I collagen degradation

F. Song, A. S. Bergdoll, L. J. Windsor

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Degenerative temporomandibular joint (TMJ) disorders are characterized by the excessive turnover of collagen. In addition to the matrix metalloproteinase (MMP) and cathepsin mediated collagen degradation pathways, a serine proteinase dependent pathway has recently been identified in TMJ fibroblasts. This study focused on further characterizing this serine proteinase pathway utilizing a media-mediated collagen degradation assay and zymography. The conditioned media from cell-mediated collagen degradation assays were incubated with Type I collagen at pH 7.5 with or without a MMP inhibitor (1,10-phenanthroline), serine proteinase inhibitors (α1-antitrypsin and soybean trypsin inhibitor, STI), or cysteine proteinase inhibitors. The data showed that 1,10-phenanthroline and STI reduced the collagen cleavage by 12.33% and 47.78%, respectively. The cysteine proteinase inhibitors had no effect. The combination of α1-antitrypsin and 1,10-phenanthroline inhibited the cleavage by 79.22%, while STI and 1,10-phenanthroline together blocked the cleavage by 85.44%. Zymography identified a proteinase at approximate 22.5 kDa, which was more effectively blocked by serine proteinase inhibitors than by MMP or cysteine proteinase inhibitors. Reverse transcript-PCR and real-time PCR results demonstrated that TMJ cells did not express trypsinogen-2 mRNA, a collagen cleaving serine proteinase. This study demonstrated that TMJ fibroblasts can predominantly utilize a serine proteinase to mediate collagen degradation, which is not trypsinogen-2.

Original languageEnglish (US)
Pages (from-to)1521-1528
Number of pages8
JournalBiochimica et Biophysica Acta - General Subjects
Volume1760
Issue number10
DOIs
StatePublished - Oct 1 2006

Keywords

  • Collagen Degradation
  • Serine Proteinase
  • Synovial Fibroblast

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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