title: The in vitro effects of dehydroepiandrosterone on human osteoarthritic chondrocytes.
abstract: OBJECTIVE
  To investigate the in vitro effects of dehydroepiandrosterone (DHEA) on human osteoarthritic chondrocytes.
  
  
  METHODS
  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.
authors.affiliation: Department of Orthopedic Surgery, Seoul National University Hospital, 28 Yongon-dong, Chongno-gu, 110-744, Seoul, South Korea.
authors.forename: H
authors.initials: H
authors.lastname: Jo
authors.forename: J S
authors.initials: JS
authors.lastname: Park
authors.forename: E M
authors.initials: EM
authors.lastname: Kim
authors.forename: M Y
authors.initials: MY
authors.lastname: Jung
authors.forename: S H
authors.initials: SH
authors.lastname: Lee
authors.forename: S C
authors.initials: SC
authors.lastname: Seong
authors.forename: S C
authors.initials: SC
authors.lastname: Park
authors.forename: H J
authors.initials: HJ
authors.lastname: Kim
authors.forename: M C
authors.initials: MC
authors.lastname: Lee
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D015153:Blotting, Western / Q000379:methods; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000276:immunology; D003094:Collagen / Q000032:analysis; D003687:Dehydroepiandrosterone / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006801:Humans; D007375:Interleukin-1 / Q000276:immunology; D020781:Matrix Metalloproteinase 1 / Q000032:analysis; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D020370:Osteoarthritis, Knee / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D014176:Protein Biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis
chemical_list: D000276:Adjuvants, Immunologic; D006025:Glycosaminoglycans; D007375:Interleukin-1; D019715:Tissue Inhibitor of Metalloproteinase-1; D003687:Dehydroepiandrosterone; D003094:Collagen; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1