Anabolic and catabolic function of chondrocyte ex vivo is reflected by the metabolic processing of type II collagen A.K. Olsen, M.Sc., B.C. Sondergaard,

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Presentation transcript:

Anabolic and catabolic function of chondrocyte ex vivo is reflected by the metabolic processing of type II collagen A.K. Olsen, M.Sc., B.C. Sondergaard, M.Sc., I. Byrjalsen, Ph.D., L.B. Tanko, M.D., Ph.D., C. Christiansen, M.D., A. Müller, Ph.D., G.E. Hein, M.D., M.A. Karsdal, Ph.D., P. Qvist, Ph.D. Osteoarthritis and Cartilage Volume 15, Issue 3, Pages 335-342 (March 2007) DOI: 10.1016/j.joca.2006.08.015 Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 The specificity of the antibody F7504 was investigated by inhibition of antibody binding to immobilized PIINP with different synthetic peptides; QDVRQPGPKG (o) and GGPKGQKGEPGDIK (⋄), both located within PIINP; QFQGPAGE (∇) located within the mature type I collagen; the immunogen (PIINP-I) conjugated to biotin at the N-terminal end (Δ), and the calibrator (PIINP-I) (□). Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Dose-dependent stimulation by IGF-I of PIINP release from bovine explants. The explants were stimulated with 1, 10 or 100ng/ml IGF-I in 5 days and PIINP was measured in the supernatant. *=P<0.05. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Bovine explants were stimulated with 10ng/ml IGF-I (●) or vehicle (○) and the PIINP (A) and CTX-II (B) were measured. The accumulation index ([PIINP]/[CTX-II]) (C) was calculated for the IGF-I stimulated explants. *=P<0.05, **=P<0.01 and ***=P<0.001. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Bovine explants were stimulated with 10ng/ml TNFα and 20ng/ml OSM (●) or vehicle (○) and the PIINP (A) and CTX-II (B) levels in the supernatant were measured. The accumulation index ([PIINP]/[CTX-II]) (C) was calculated for the TNFα and OSM stimulated explants. *=P<0.05 and ***=P<0.001. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 BMP-2 levels were determined in the supernatants of OSM and TNFα stimulated bovine cartilage explants (gray bars) or vehicle (black bars), and expressed as the average concentration in the early phase (days 0–9) and late phase (days 12–19), respectively. T=TNFα; n.s.=none significant. *=P<0.05. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 MMP dependent release of PIINP (A) and CTX-II (B) was investigated in the presence or absence of broad spectrum MMP inhibitor, GM6001. The proteases were incubated for 24h with metabolic inactivated cartilage. For comparison, markers were determined in bovine SF. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 Serum PIINP (A) and urinary CTX-II (B) were determined in healthy subjects (n=15) and in patients with RA (n=15). In addition, the Z-scores (C) for PIINP, CTX-II and accumulation index in 15 patients with RA were calculated. Lines represented the mean Z-score. For calculation of Z-scores please refer to Statistical analysis. Osteoarthritis and Cartilage 2007 15, 335-342DOI: (10.1016/j.joca.2006.08.015) Copyright © 2006 Osteoarthritis Research Society International Terms and Conditions