Changes of early post-traumatic osteoarthritis in an ovine model of simulated ACL reconstruction are associated with transient acute post-injury synovial.

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Changes of early post-traumatic osteoarthritis in an ovine model of simulated ACL reconstruction are associated with transient acute post-injury synovial inflammation and tissue catabolism B.J. Heard, N.M. Solbak, Y. Achari, M. Chung, D.A. Hart, N.G. Shrive, C.B. Frank Osteoarthritis and Cartilage Volume 21, Issue 12, Pages 1942-1949 (December 2013) DOI: 10.1016/j.joca.2013.08.019 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Gross morphological grading of 2 and 20 week animals. Both 2 and 20 week joints had significantly greater scores in their surgical joints than their respective contralaterals. (A) Paired comparison of right (surgical) and left (contralateral) joints (Wilcoxon matched pairs signed rank test, * = P ≤ 0.05, ** = P ≤ 0.01). (B) Treatment group comparison of adjusted values (surgical joint score – contralateral joint score) (mean ± standard deviation) (Kruskal Wallis test with Dunn's multiple comparisons test), bar + (* = P ≤ 0.05, ** = P ≤ 0.01, *** = P ≤ 0.001). 2 and 20 week ACL-R adjusted values were significantly greater than those of the normal group, with 20 week ACL-R also being significantly greater than 20 week sham. Osteoarthritis and Cartilage 2013 21, 1942-1949DOI: (10.1016/j.joca.2013.08.019) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Histological grading of synovial inflammation. All data are represented as adjusted scores (surgical joint – contralateral joint values). (A) Treatment group comparison of combined (all four criteria) histology score (mean ± standard deviation). Although variable, a trend suggests that inflammation is greater within the 2 week ACL-R animals. No statistical differences (P > 0.05). (B) Breakdown of the contributions of each inflammatory criterion scored. IH = intimal hyperplasia, LI = lympho/plasmo-cytic infiltration, SIF = subintimal fibrosis, and V = vascularity. (Tukey's box and whiskers plot – dots are outliers) No statistical differences (P > 0.05). Osteoarthritis and Cartilage 2013 21, 1942-1949DOI: (10.1016/j.joca.2013.08.019) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 (A) Synovial mRNA expression. Synovial tissue samples from the 2 week time point were reexamined alongside samples from the 20 week time point. The mRNA expression levels for IL-1β and MMP 13 were significantly elevated in paired comparisons between 2 week ACL-R surgical and contralateral values (left side of each graph) (Wilcoxon matched pairs signed rank test, ** = P ≤ 0.01). Adjusted (surgical joint – contralateral joint values) mRNA expression levels for IL-1β and MMP 13 were significantly elevated in 2 week ACL-R when compared to 20 week ACL-R (right side of each graph: Tukey's box and whiskers plot – dots are outliers) (Kruskal Wallis test with Dunn's multiple comparisons test, bar = P ≤ 0.05). (B) Summary of synovial protein expression (western blot with band normalized to GAPDH) (mean ± standard deviation). IL-1β appears to be present in the highest concentration in 2 week ACL-R samples. MMP 13 appears to be present in both 2 and 20 week ACL-R surgical joint samples. Osteoarthritis and Cartilage 2013 21, 1942-1949DOI: (10.1016/j.joca.2013.08.019) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

S-Table I Expression of mRNA for select inflammatory cytokines, catabolic, and anabolic molecules at 2 and 20 weeks after ACL-R surgery (mean and standard deviation values from stifle joints of the right limbs only). ACL-R (n = 7), Sham (n = 7), and Normal (n = 8). Osteoarthritis and Cartilage 2013 21, 1942-1949DOI: (10.1016/j.joca.2013.08.019) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions