Lentiviral shRNA knock-down of ADAMTS-5 and -9 restores matrix deposition in 3D chondrocyte culture. Academic Article uri icon

Overview

abstract

  • Aggrecan is one of the two major constituents of articular cartilage, and during diseases such as osteoarthritis (OA) it is subject to degradation by proteolytic enzymes. The primary proteases responsible for aggrecan cleavage are the aggrecanases, identified as members of the ADAMTS family of proteases, which are upregulated in response to inflammatory stimuli. It is uncertain which of the six aggrecanases (ADAMTS-1, -4, -5, -8, -9 and -15) are primarily responsible for the degradation of aggrecan in human cartilage. Here we show that four of the six aggrecanases are expressed in immortalized chondrocyte cell-lines and can be upregulated in response to inflammatory cytokines. Using RNA interference, we demonstrate robust knock-down of ADAMTS-5 and -9 expression in these cells and, by culturing them on three-dimensional (3D) scaffolds, show that reduction in expression of ADAMTS-5 enzyme results in an increase in matrix deposition. These data suggest that the quality of tissue-engineered cartilage matrix might be improved by targeted depletion of aggrecanase expression. Moreover, this work also provides further evidence that ADAMTS-5 may be a therapeutic target in the treatment of arthritic disease.

publication date

  • December 1, 2010

Research

keywords

  • ADAM Proteins
  • Chondrocytes
  • Extracellular Matrix
  • Gene Knockdown Techniques
  • Lentivirus
  • RNA, Small Interfering

Identity

PubMed Central ID

  • PMC2988096

Scopus Document Identifier

  • 78649677599

Digital Object Identifier (DOI)

  • 10.1002/term.275

PubMed ID

  • 20568084

Additional Document Info

volume

  • 4

issue

  • 8