The Effects of Dehydroepiandrosterone-pyruvate on Human Osteoarthritic Chondrocytes

Lee, Myung Chul; Ji, Jong Hun; Kim, Weon Yoo; Do, Jeong Hun; Kim, Yong Min

Abstract

PURPOSE
To investigate the in vitro effects of Dehydroepiandrosterone (DHEA)-pyruvate on human osteoarthritic chondrocytes.
METHODS
Chondrocytes isolated from human osteoarthritic knee cartilage were three-dimensionally cultured in Alginate beads. Cells were treated with dehydroepiandrosterone in the presence or absence of IL-1. The effects on chondrocytes were analyzed by MTS assay (for chondrocytes proliferation), DMB assay (for glycosaminoglycan synthesis), and indole assay (for DNA amount). Gene expressions of MMP-1,3, TIMP-1 as well as IL-1 induced gene expression of MMP-1, 3 were analyzed using RT-PCR. The protein synthesis of MMP-1,3 and TIMP-1 was determined by ELISA.
RESULTS
Treatment of chondrocytes with DHEA-pyruvate did not affect chondrocytes proliferation regardless of concentrations when compared with control. GAG synthesis was not changed significantly during culture period regardless of concentrations. DHEA-pyruvate suppressed the expression of MMP-1 significantly at a concentration of 50 micrometer and above. The gene expression of MMP-3 was also suppressed. The expression of TIMP-1 was significantly increased by DHEA-pyruvate at concentration of 50 micrometer. The effects of DHEA-pyruvate were also prominent in the presence of IL-1, in which IL-1induced gene expressions of not only MMP-1, but also MMP-3 were suppressed at a lower concentration of 10 micrometer and 50 micrometer, respectively. In enzyme activity measurement, the results came in line with the results obtained by RT-PCR, which means DHEA-pyruvate influences on the gene transcription level.
CONCLUSION
Our study clearly demonstrated that DHEA-pyruvate has the ability to modulate the imbalance between MMPs and TIMP-1 during osteoarthritis at the transcription level, suggesting its protective role against loss of articular cartilage.