Cartilage and bone changes during development of post-traumatic osteoarthritis in selected LGXSM recombinant inbred mice S. Hashimoto, M.F. Rai, K.L.

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Cartilage and bone changes during development of post-traumatic osteoarthritis in selected LGXSM recombinant inbred mice S. Hashimoto, M.F. Rai, K.L. Janiszak, J.M. Cheverud, L.J. Sandell Osteoarthritis and Cartilage Volume 20, Issue 6, Pages 562-571 (June 2012) DOI: 10.1016/j.joca.2012.01.022 Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Ear wound healing in LGXSM-6 and LGXSM-33 strains. Through-and-through ear puncture wounds (2-mm in diameter) on external ears of LGXSM-6 and LGXSM-33 strains were created at 6 weeks of age using standard method. Gross appearance of ear hole in LGXSM-6 (A) and LGXSM-33 (B) mice after 30 days of ear punch is shown. A large ear hole retained in LGXSM-33 strain indicates a failure of healing as compared to LGXSM-6 strain. A graphical representation of residual ear wound diameter (C) after 15 and 30 days of ear punch showed that LGXSM-6 mice significantly healed their ears at both day 15 and day 30 compared to LGXSM-33 mice. Osteoarthritis and Cartilage 2012 20, 562-571DOI: (10.1016/j.joca.2012.01.022) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Histological evaluation for OA development. Representative histological sections of LGXSM-6 (A) and LGXSM-33 (B) mice after 2, 4 and 8 weeks of DMM surgery (×40) are shown. Black arrows show cartilage damage in medial tibial plateau. MFC=medial femoral condyle, LFC=lateral femoral condyle, MTP=medial tibial plateau, LTP=lateral tibial plateau, MM=medial meniscus, LM=lateral meniscus. Osteoarthritis and Cartilage 2012 20, 562-571DOI: (10.1016/j.joca.2012.01.022) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 OA score for LGXSM-6 and LGXSM-33. The sum OA score (A) from all the four quadrants and eight sections from each knee was based on cartilage damage and shows that LGXSM-33 strain overall develops more OA compared to LGXSM-6 after 8 weeks of surgery. A maximum score (B) representing highest score from medial compartment of LGXSM-6 and LGXSM-33 indicates that DMM knees had significant more OA score in both strains compared to sham knee. It also shows that LGXSM-33 has significantly higher grade of OA than LGXSM-6 at 8 weeks post-surgery Filled circles =individual data points, Star = mean value, hyphens = upper and lower limits of 95% CI. Osteoarthritis and Cartilage 2012 20, 562-571DOI: (10.1016/j.joca.2012.01.022) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Changes in subchondral bone thickness in LGXSM-33 and LGXSM-6 strains. Coronal sectional images were used for measurement of subchondral bone thickness in the medial and lateral tibial plateau (A). Subchondral bone thickness on medial and lateral sides of LGXSM-33 after 2, 4, and 8 weeks of DMM surgery showed that there were significant differences in thickness of subchondral bone between sham and DMM on medial side after 8 weeks. Similarly, the DMM knees at 8 weeks from both medial and lateral sides were significantly higher than 2 and 4 weeks. In contrast, in LGXSM-6 mice the differences between subchondral bone thickness in sham and DMM knee varied significantly at 2 and 4 weeks on medial side and only at 4 weeks on lateral side. The overall data from A and B show that LGXSM-33 develops more subchondral bone thickness than LGXSM-6 Filled circles = individual data points, Star = mean value, hyphens = upper and lower limits of 95% CI. Osteoarthritis and Cartilage 2012 20, 562-571DOI: (10.1016/j.joca.2012.01.022) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Trabecular bone parameters for LGXSM-6 and LGXSM-33 strains. All parameters were derived from trabecular bone of the entire epiphysis of DMM and sham in both strains. Bone volume fraction (BV/TV) (A), trabecular thickness (B) and connectivity density index (C) are shown for LGXSM-6 and LGXSM-33 strains Filled circles =individual data points, Star = mean value, hyphens = upper and lower limits of 95% CI. Osteoarthritis and Cartilage 2012 20, 562-571DOI: (10.1016/j.joca.2012.01.022) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions