Analysis of radial variations in material properties and matrix composition of chondrocyte-seeded agarose hydrogel constructs T.-A.N. Kelly, Ph.D., K.W.

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Analysis of radial variations in material properties and matrix composition of chondrocyte-seeded agarose hydrogel constructs T.-A.N. Kelly, Ph.D., K.W. Ng, Ph.D., G.A. Ateshian, Ph.D., C.T. Hung, Ph.D. Osteoarthritis and Cartilage Volume 17, Issue 1, Pages 73-82 (January 2009) DOI: 10.1016/j.joca.2008.05.019 Copyright © 2008 Terms and Conditions

Fig. 1 Images of free-swelling (A, B) and dynamically loaded (C, D) constructs were acquired at day 42, before (A, C) and after (B, D) coring of the constructs. Scale bars equal 5mm. For this study, samples were tested whole, allowed to recover and then the annuli and cores were tested separately (E). Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 2 Wet and dry weights (A), water content (B), DNA content (C, D), and GAG and collagen contents (E, F) of intact, free-swelling and dynamically loaded chondrocyte-seeded constructs. *Represents significant differences vs day 0 constructs (ANOVA P<0.005; post hoc P<0.005). Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 3 Wet weight (A), dry weight (B), water content (C), and DNA content (D, E), of the annuli (open) and cores (closed) of free-swelling (boxes) and dynamically loaded (triangles) constructs over the 6-week culture period (n=4–9). Significant effect of time was observed for these samples (ANOVA P<0.05). *Represents significant differences vs day 0 constructs (ANOVA P<0.05; post hoc P<0.05); †represents significant differences vs free-swelling controls (ANOVA P<0.01; post hoc P<0.05); §represents significant differences vs the respective cores (ANOVA P<0.001; post hoc P<0.05). Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 4 GAG (A, B) and collagen (C, D) contents of the annuli (open) and cores (closed) of free-swelling (boxes) and dynamically loaded (triangles) constructs over the 6-week culture period (n=4–9). Significant effect of time was observed for these samples (ANOVA P<0.05). *Represents significant differences vs day 0 constructs (ANOVA P<0.05; post hoc P<0.05); †represents significant differences vs free-swelling controls (ANOVA P<0.05; post hoc P<0.05); §represents significant differences vs the respective cores (ANOVA P<0.05; post hoc P<0.05). Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 5 Equilibrium (A) and dynamic (B; measured at 0.5Hz) moduli of free-swelling and dynamically loaded constructs over the 42-day culture period (n=7). The constructs were allowed to recover for 1h before the annulus and the 3mm central core were tested. As the dynamic modulus is a structural property, inter-group comparisons between intact constructs (B), annuli (C), and cores (D) were not performed. *Represents significant differences compared to day 0 (ANOVA P<0.005; Tukey P<0.003); ‡represents significant differences compared to corresponding free-swelling controls (ANOVA P<0.0005; Tukey P<0.005); †represents significant differences compared to whole sample (ANOVA P<0.0005; Tukey P<0.02); §represents significant differences compared to day 28 constructs (P<0.05). Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 6 Safranin O (GAG; A–F) and Picrosirius Red staining (collagen; G–L) of free-swelling (A–C, G–I) and dynamically loaded (D–F, J–L) constructs on day 42. Images were acquired along the radial edge (first column), in the center (second column), and along the axial edge (third column) of the constructs. Scale bars equal 250μm. Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions

Fig. 7 Immunofluorescent labeling (green) for type II collagen (A–F), type I collagen (G–L), and elastin (M–R) of free-swelling (A–C, G–I, M–O) and dynamically loaded (D–F, J–L, P–R) chondrocyte-seeded constructs at day 42. Images were acquired along the radial edge (first column), in the center (second column), and along the axial edge (third column) of the constructs. Propidium iodide counter staining (red) was used to visualize the cell nuclei. Scale bars equal 200μm. Osteoarthritis and Cartilage 2009 17, 73-82DOI: (10.1016/j.joca.2008.05.019) Copyright © 2008 Terms and Conditions