Vitamin D Regulation of Metal loproteinase Activity in Matrix Vesicles

David D. Dean, Zvi Schwartz, John Schmitz, Ofelia E. Muniz, Yuhun lu, Felix Calderon, David S. Howell, Barbara D. Boyan

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

Matrix vesicles (MVs) are enriched in matrix metalloproteinases (MMPs) capable of degrading proteoglycans. The aim of the present study was to identify which MMPs are present in MVs and determine whether these MMPs are regulated by l, 25-(OH)2D3 [1, 25] and 24, 25-(OH)2D3 [24, 25]. To do this, growth zone (GC) and resting zone (RC) chondrocytes were isolated from rat costochondral cartilage and placed into culture. At confluence, GCs were treated with 1, 25 and RCs with 24, 25 for 24 hours. MVs, plasma membranes (PMs), and conditioned media were then collected from the cultures. RT-PCR demonstrated the presence of mRNA for stromelysin-1 and 72 kDa gelatinase in both RCs and GCs. Casein zymography revealed activity at Mr 48 and 28 kDa in MV, but not PM or conditioned media; Western analysis confirmed that this activity was stromelysin-1. Gelatinolytic activity, at low levels, was also found in MVs, but not PMs or conditioned media. When enzyme activity was measured using a proteoglycan bead assay, it was found that both GCs and RCs produced MVs and PMs containing neutral metalloproteinase. Both cells also produced MVs and PMs containing plasminogen activator. The addition of 1, 25 to GCs caused a significant 4- to 5-fold increase in metalloproteinase activity in MVs, but not PMs. In contrast, MVs from cultures of RCs treated with 24, 25 contained decreased metalloproteinase activity; enzyme activity in PMs was unaffected by 24, 25. Plasminogen activator in MVs from RC was increased by treatment with 24, 25, while MV enzyme activity was decreased after treatment of GC cultures with 1, 25. This study shows that both RCs and GCs produce stromelysin-1 and 72 kDa gelatinase and that these enzymes are preferentially localized in MVs. Further, MMP and plasminogen activator activities in MVs and PMs are regulated by vitamin D metabolites.

Original languageEnglish (US)
Pages (from-to)331-336
Number of pages6
JournalConnective Tissue Research
Volume35
Issue number42008
DOIs
Publication statusPublished - Jan 1 1996

    Fingerprint

Keywords

  • 1, 25-dihydroxyvitamin
  • D, 24, 25-dihydroxyvitamin D
  • calcification
  • casein zymography

ASJC Scopus subject areas

  • Rheumatology
  • Biochemistry
  • Orthopedics and Sports Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Dean, D. D., Schwartz, Z., Schmitz, J., Muniz, O. E., lu, Y., Calderon, F., ... Boyan, B. D. (1996). Vitamin D Regulation of Metal loproteinase Activity in Matrix Vesicles. Connective Tissue Research, 35(42008), 331-336. https://doi.org/10.3109/03008209609029208