Journal
JOURNAL OF BIOMECHANICS
Volume 42, Issue 13, Pages 2177-2182Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.jbiomech.2009.05.039
Keywords
Chondrocytes; Cartilage; Tissue engineering; Mechanical stimulation; Mechanotransduction; Gene expression
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Funding
- German Ministry of Education and Research [0313836]
- German Research Foundation [BA 1025/2-1]
- Dutch Technology Foundation STW [VIDI-07970]
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Recent studies have shown that integrins act as mechanoreceptors in articular cartilage. In this study, we examined the effect of blocking RGD-dependent integrins on both ECM gene expression and ECM protein synthesis. Chondrocytes were isolated from full-depth porcine articular cartilage and seeded in 3% agarcise constructs. These constructs were loaded in compression with 15% strain at 0.33 and 1 Hz for 12 h, in the presence or absence of GRGDSP, which blocks RGD-dependent integrin receptors. The levels of mRNA for aggrecan, collagen II and MMP-3 were determined by semi-quantitative PCR at several time points up to 24 h post-stimulation. DNA and sGAG content were determined at several time points up to 28 days post-stimulation. At 0.33 Hz, the mRNA levels for aggrecan and MMP-3 were increased after loading, but the mRNA levels for collagen II remained unchanged. Incubation with GRGDSP counteracted these effects. Loading at 1 Hz led to increased mRNA levels for all three molecules directly after loading and these effects were counteracted by incubation with GRGDSP. The constructs that were loaded at 0.33 Hz showed a lower amount of sGAG, compared to the unstrained control. In contrast, loading at 1 Hz caused an increase in sGAG deposition over the culture period. Blocking integrins had only a counteracting effect on the long-term biosynthetic response of constructs that were compressed at 1 Hz. The results confirmed the role of RGD-dependent integrins as mechanotransducers in the regulation of both ECM gene expression and matrix biosynthesis for chondrocytes seeded in agarose under the applied loading regime. Interestingly, this role seems to be dependent on the applied loading frequency. (C) 2009 Elsevier Ltd. All rights reserved.
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