TY - JOUR
T1 - RGD-dependent integrins are mechanotransducers in dynamically compressed tissue-engineered cartilage constructs
AU - Kock, L.M.
AU - Schulz, R.M.
AU - Donkelaar, van, C.C.
AU - Thümmler, C.B.
AU - Bader, A.
AU - Ito, K.
PY - 2009
Y1 - 2009
N2 - 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% agarose 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.
AB - 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% agarose 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.
U2 - 10.1016/j.jbiomech.2009.05.039
DO - 10.1016/j.jbiomech.2009.05.039
M3 - Article
C2 - 19656515
SN - 0021-9290
VL - 42
SP - 2177
EP - 2182
JO - Journal of Biomechanics
JF - Journal of Biomechanics
IS - 13
ER -