12397608

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3

Hydrostatic pressure (HP) is thought to increase within cartilage extracellular matrix as a consequence of fluid flow inhibition. The biosynthetic response of human articular chondrocytes to HP in vitro varies with the load magnitude, load frequency, as well as duration of loading. We found that continuous cyclic HP (5 MegaPascals (MPa) for 4 h; 1 Hz frequency) induced apoptosis in human chondrocytes derived from osteoarthritic cartilage in vitro as evidenced by reduced chondrocyte viability which was independent of initial cell densities ranging from 8.1 x 10(4) to 1.3 x 10(6) cells ml(-1). HP resulted in internucleosomal DNA fragmentation, activation of caspase-3, and cleavage of poly-ADP-ribose polymerase (PARP). At the molecular level, induction of apoptosis by HP was characterized by up-regulation of p53, c-myc, and bax-alpha after 4 h with concomitant down-regulation of bcl-2 after 2 h at 5 MPa as measured by RT-PCR. In contrast, beta-actin expression was unchanged. Real-time quantitative RT-PCR confirmed a HP-induced (5 MPa) 1.3-2.6 log-fold decrease in bcl-2 mRNA copy number after 2 and 4 h, respectively, and a significant increase (1.9-2.5 log-fold) in tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) mRNA copy number after 2 and 4 h, respectively. The up-regulation of p53 and c-myc, and the down-regulation of bcl-2 caused by HP were confirmed at the protein level by Western blotting. These results indicated that HP is a strong inducer of apoptosis in osteoarthritic human chondrocytes in vitro.

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pubmed-author:GoldbergVictor MVM, pubmed-author:HaqqiTariq MTM, pubmed-author:IslamNajmulN, pubmed-author:JepsenKarl JKJ, pubmed-author:KraayMatthewM, pubmed-author:MalemudCharles JCJ, pubmed-author:WelterJean FJF

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266-78

pubmed-meshheading:12397608-Apoptosis, pubmed-meshheading:12397608-Cartilage, Articular, pubmed-meshheading:12397608-Caspase 3, pubmed-meshheading:12397608-Caspases, pubmed-meshheading:12397608-Cell Survival, pubmed-meshheading:12397608-Chondrocytes, pubmed-meshheading:12397608-DNA Primers, pubmed-meshheading:12397608-Down-Regulation, pubmed-meshheading:12397608-Enzyme Precursors, pubmed-meshheading:12397608-Humans, pubmed-meshheading:12397608-Hydrostatic Pressure, pubmed-meshheading:12397608-Nitric Oxide Synthase, pubmed-meshheading:12397608-Nitric Oxide Synthase Type II, pubmed-meshheading:12397608-Osteoarthritis, pubmed-meshheading:12397608-Poly(ADP-ribose) Polymerases, pubmed-meshheading:12397608-Proto-Oncogene Proteins, pubmed-meshheading:12397608-Proto-Oncogene Proteins c-bcl-2, pubmed-meshheading:12397608-Proto-Oncogene Proteins c-myc, pubmed-meshheading:12397608-Tumor Necrosis Factor-alpha, pubmed-meshheading:12397608-Tumor Suppressor Protein p53, pubmed-meshheading:12397608-Up-Regulation, pubmed-meshheading:12397608-bcl-2-Associated X Protein

Hydrostatic pressure induces apoptosis in human chondrocytes from osteoarthritic cartilage through up-regulation of tumor necrosis factor-alpha, inducible nitric oxide synthase, p53, c-myc, and bax-alpha, and suppression of bcl-2.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't