The in vitro effects of dehydroepiandrosterone on human osteoarthritic chondrocytes

Osteoarthritis Cartilage. 2003 Aug;11(8):585-94. doi: 10.1016/s1063-4584(03)00094-3.


Objective: To investigate the in vitro effects of dehydroepiandrosterone (DHEA) on human osteoarthritic chondrocytes.

Design: Chondrocytes isolated from human osteoarthritic knee cartilage were three-dimensionally cultured in alginate beads, except for cell proliferation experiment. Cells were treated with DHEA in the presence or absence of IL-1beta. The effects on chondrocytes were analyzed using a 3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay (for chondrocyte proliferation), a dimethylmethylene blue (DMB) assay (for glycosaminoglycan (GAG) synthesis), and an indole assay (for DNA amount). Gene expressions of type I and II collagen, metalloproteinase-1 and -3 (MMP-1 and -3), and tissue inhibitor of metalloproteinase-1 (TIMP-1) as well as the IL-1beta-induced gene expressions of MMP-1 and -3 were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The protein synthesis of MMP-1 and -3 and TIMP-1 was determined by Western blotting.

Results: The treatment of chondrocytes with DHEA did not affect chondrocyte proliferation or GAG synthesis up to 100 micro M of concentration. The gene expression of type II collagen increased in a dose-dependent manner, while that of type I decreased. DHEA suppressed the expression of MMP-1 significantly at concentrations exceeding 50 micro M. The gene expression of MMP-3 was also suppressed, but this was without statistical significance. The expression of TIMP-1 was significantly increased by DHEA at concentrations exceeding 10 micro M. The effects of DHEA on the gene expressions of MMP-1 and -3 were more prominent in the presence of IL-1beta, in which DHEA suppressed not only MMP-1, but also MMP-3 at the lower concentrations, 10 and 50 micro M, respectively. Western blotting results were in agreement with RT-PCR, which indicates that DHEA acts at the gene transcription level.

Conclusions: Our study demonstrates that DHEA has no toxic effect on chondrocytes up to 100 micro M of concentration and has an ability to modulate the imbalance between MMPs and TIMP-1 during OA at the transcription level, which suggest that it has a protective role against articular cartilage loss.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic / pharmacology*
  • Blotting, Western / methods
  • Cell Division / drug effects
  • Cells, Cultured
  • Chondrocytes / drug effects*
  • Chondrocytes / immunology
  • Collagen / analysis
  • Dehydroepiandrosterone / pharmacology*
  • Gene Expression / drug effects
  • Glycosaminoglycans / biosynthesis
  • Humans
  • Interleukin-1 / immunology
  • Matrix Metalloproteinase 1 / analysis
  • Matrix Metalloproteinase 3 / analysis
  • Osteoarthritis, Knee / immunology
  • Osteoarthritis, Knee / metabolism
  • Osteoarthritis, Knee / pathology*
  • Protein Biosynthesis
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Tissue Inhibitor of Metalloproteinase-1 / analysis


  • Adjuvants, Immunologic
  • Glycosaminoglycans
  • Interleukin-1
  • Tissue Inhibitor of Metalloproteinase-1
  • Dehydroepiandrosterone
  • Collagen
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 1