Basic calcium phosphate crystals up-regulate metalloproteinases but down-regulate tissue inhibitor of metalloproteinase-1 and -2 in human fibroblasts

G. Bai, D. S. Howell, G. A. Howard, B. A. Roos, H. S. Cheung

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

Objective: To examine the effect of basic calcium phosphate (BCP) crystals on expression of tissue inhibitors of metalloproteinases (TIMP)-1 and -2 in human fibroblasts. Method: Using a semi-quantitative reverse transcription-polymerase chain reaction method and phosphocitrate (PC), a specific inhibitor of the biological effects of BCP crystals, we examined the effects of BCP on the steady state transcript levels of metalloproteinase (MMP)-1, -3, -9 and -13 and TIMP-1 and -2 in human fibroblasts. DNA primers against elongation factor were used as internal controls. RNAs isolated from human fibroblasts treated with BCP crystals (50 μg/ml) in the presence or absence of PC (10-3 M) were used as templates, and RNA from untreated control cultures and cultures treated with Interleukin-1-beta (IL-1β) were used as negative and positive controls, respectively. Results: We observed increases in MMP-1, -3, -9 and -13 transcripts by BCP crystals. BCP crystal down-regulated TIMP-1 and -2 over untreated controls. Western blot analysis confirmed that BCP crystals down-regulate the synthesis of TIMP-1 and -2. While IL-1β up-regulated MMP-1, -3, -9 and -13, it had no significant effect on expression of either TIMP. In all cases, PC specifically reversed the differential regulation of MMPs and TIMPs by BCP crystals but had no effect on IL-1β induction of MMP expression. Conclusion: The ability of BCP to induce the synthesis of degradative MMPs while down-regulating the synthesis of the naturally occurring counterpart TIMPs may explain the changes consistent with a role of BCP crystal in the pathogenesis of degenerative changes in osteoarthritis. The ability of PC to reverse both degradative effects of BCP crystal suggests that PC can be a potential therapeutic agent for BCP crystal deposition diseases.

Original languageEnglish (US)
Pages (from-to)416-422
Number of pages7
JournalOsteoarthritis and Cartilage
Volume9
Issue number5
DOIs
StatePublished - Jul 2001

Keywords

  • BCP Crystal
  • MMP
  • PC
  • RT-PCR
  • TIMP

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine

Fingerprint

Dive into the research topics of 'Basic calcium phosphate crystals up-regulate metalloproteinases but down-regulate tissue inhibitor of metalloproteinase-1 and -2 in human fibroblasts'. Together they form a unique fingerprint.

Cite this