Y. M. Bastiaansen-Jenniskens, M. Sc. , W. Koevoet, B. Sc. , A. C. W

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Presentation transcript:

Contribution of collagen network features to functional properties of engineered cartilage Y.M. Bastiaansen-Jenniskens, M.Sc., W. Koevoet, B.Sc., A.C.W. de Bart, B.Sc., J.C. van der Linden, Ph.D., A.M. Zuurmond, Ph.D., H. Weinans, Ph.D., J.A.N. Verhaar, M.D., Ph.D., G.J.V.M. van Osch, Ph.D., J. DeGroot, Ph.D. Osteoarthritis and Cartilage Volume 16, Issue 3, Pages 359-366 (March 2008) DOI: 10.1016/j.joca.2007.07.003 Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Effect of (transient) collagen crosslink inhibition on collagen crosslinking (A) and collagen deposition (B) during 10 weeks of culture (n=9). Collagen crosslinking was inhibited by BAPN until day 35 in the +/− condition. From day 35 onwards, two conditions were cultured without the presence of BAPN and crosslinks could be formed. In one condition, crosslinks were inhibited during the full 70 days of culture. * Indicates P<0.002 between the control and the BAPN +/− conditions at the same time point. † Indicates P<0.001 between BAPN +/+ and the other two conditions. ‡ Indicates P<0.005 between BAPN +/+ and the other two conditions. Data are shown as mean±SD. Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Mechanical properties of alginate discs cultured for 5 weeks (A&B) and 10 weeks (C&D). Both equilibrium aggregate modulus (A&C) and secant modulus (B&D) were examined (n=8). * Indicates P<0.002 compared to the control condition without inhibition of crosslink formation. Data are presented as mean modulus±SD. Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Susceptibility of the matrix for MMP-1 degradation after 10 weeks of culture. Amount of collagen in the soluble fraction directly after incubation with 100nM MMP-1 is higher for the matrix without sufficient collagen crosslinks as represented by the +/+ condition than for matrices with collagen crosslinks (−/− and +/−), even when crosslink formation was transiently inhibited (+/−). * Indicates P<0.001 compared to the −/− and +/− conditions. Data are presented as mean percentage of the total amount of collagen in the beads±SD (n=6). Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Distribution of collagen between the CM and the FRM. No significant differences are found in distribution between the three conditions. Data are shown as mean percentage per compartment±SD (n=6). Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Gene expression of COLI, COLII and SOX9, shown as COLII–COLI gene expression ratio (A) and relative SOX9 gene expression (B). Both parameters are indicative for the phenotype of the chondrocytes. * Indicates P<0.001 compared to the −/− and +/− conditions. Data are presented as mean±SD (n=3). Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Immunocytochemical analysis of produced matrix after 35 days of culture. Control chondrocytes stained for COLII (A) and COLI (B) and chondrocytes cultured in the presence of BAPN for 35 days stained for COLII (C) and COLI (D). Magnification is 100 times, with a 400 times inset. Positive cells are indicated in red with haematoxylin counter stain (purple). Osteoarthritis and Cartilage 2008 16, 359-366DOI: (10.1016/j.joca.2007.07.003) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions