M.Z.C. Ruan, R.M. Patel, B.C. Dawson, M.-M. Jiang, B.H.L. Lee

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Pain, motor and gait assessment of murine osteoarthritis in a cruciate ligament transection model M.Z.C. Ruan, R.M. Patel, B.C. Dawson, M.-M. Jiang, B.H.L. Lee Osteoarthritis and Cartilage Volume 21, Issue 9, Pages 1355-1364 (September 2013) DOI: 10.1016/j.joca.2013.06.016 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Experiment scheme of functional change evaluation. We performed functional evaluation when the mice were 7 months old. This was followed by CLT and sham surgery. In different batches of mice, we waited 4, 8 or 12 weeks before another functional assessment. We performed postmortem assessment after the second and final functional analysis. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Pain sensitivity and mobility measured by hotplate analysis. (a) Experiment setup and device of hotplate analysis; (b) Response time of the mice before and at different time points after surgery; (c) Performance change on hotplate normalized to individual mice. Performance change = [(timeafter − timebefore)/timebefore] × 100%. N = 12-14. All error bars represent s.e.m. *P < 0.05, **P < 0.01. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Movement of mice measured by open field analysis (OFA). (a) Arena of the open field analysis; (b) Total distance traveled (during testing period); (c) Average walking speed of the mice in the open field (speed = total distance travelled/total moving time); (d) Total rest time; (e) Number of rearing movement; (f) Average time of each rearing event. N = 10. All error bars represent s.e.m. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Motor function of mice measured by Rotarod analysis. (a) Experiment setup of Rotarod analysis; (b–d) First ride around time of the mice before the procedure (trials 1–6) and 4 weeks (b, n = 20–23), 8 weeks (c, n = 19–20) and 12 weeks (d, n = 17–18) after either sham or CLT; (e) Performance change of average Rotarod first ride around time of trials performed before and after surgery. Performance change = [(timeafter − timebefore)/timebefore] × 100%. All error bars represent s.e.m. *P < 0.05, **P < 0.01. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Gait change of mice with osteoarthritis induced by CLT. (a) Experiment setup of gait analysis; (b–d) Stride and base of the mice 4 weeks (b), 8 weeks (c) and 12 weeks (d) after surgery; (e) Representative hind limb foot prints of Sham and CLT mice on gait analysis; (f) Quantification of hind limb foot print area at different time points after surgery. N = 17-19. All error bars represent s.e.m. All the significant comparisons are labeled, *P < 0.05, **P < 0.01. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Longitudinal evaluation of cartilage loss and pathological changes in CLT mice. (a) Cartilage surface area, (b) Cartilage volume (c) subchondral bone area covered by cartilage (d) osteophyte number and (e) osteophyte volume in whole hind limb knee joint in mice at different time points after surgery, N = 5–6; (f) Maximum OARSI score on sham and CLT whole hind limb joints in mice at different time points after surgery, N = 8. (g–i) Synovitis score at 4 weeks (g), 8 weeks (h) and 12 weeks (i) post surgery at medial femur (MF), medial tibia (MT), lateral femur (LF) and lateral tibia (LT), N = 5. (j–l) Bivariate correlation between OARSI score and performance changes in rotarod analysis (Spearman rho = −0.6024, P < 0.0001, n = 39) (j), hotplate analysis (Spearman rho = 0.3585, P < 0.05, n = 34) (k) and hind limb foot print area (Spearman rho = −0.4586, P < 0.05, n = 29) (l) All error bars represent s.e.m. *P < 0.05, **P < 0.01. Osteoarthritis and Cartilage 2013 21, 1355-1364DOI: (10.1016/j.joca.2013.06.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions