In vivo imaging of MMP-13 activity in the murine destabilised medial meniscus surgical model of osteoarthritis N.H. Lim, E. Meinjohanns, M. Meldal, G.

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In vivo imaging of MMP-13 activity in the murine destabilised medial meniscus surgical model of osteoarthritis N.H. Lim, E. Meinjohanns, M. Meldal, G. Bou-Gharios, H. Nagase Osteoarthritis and Cartilage Volume 22, Issue 6, Pages 862-868 (June 2014) DOI: 10.1016/j.joca.2014.04.006 Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Histological progression of DMM in this study. Typical histological sections of the DMM operated right knee joints stained with Safranin O obtained at (A) 4 weeks, (B) 6 weeks and (C) 8 weeks after surgery. Damaged areas within cartilage sections are indicated with arrows. (D) Summed histological scores obtained from addition of the scores of the top 3 consecutive scoring slides per knee. Data are mean ± 95% CI. Osteoarthritis and Cartilage 2014 22, 862-868DOI: (10.1016/j.joca.2014.04.006) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Representative images obtained 4 h following intravenous injection of MMP12ap and MMP13ap in mice following sham and DMM surgery. Typical X-ray and fluorescence images obtained 4 h following intravenous injection of either 150 μl of 1 μM MMP12ap or MMP13ap after 4, 6 and 8 weeks after DMM or sham surgery. Left knees (LK), right knees (RK) and the red circle marking the ROI used for analysis are indicated. The fluorescence rainbow scales are indicated on the right. Osteoarthritis and Cartilage 2014 22, 862-868DOI: (10.1016/j.joca.2014.04.006) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Quantification of the activation of MMP12ap in the DMM model of OA. Mice were injected with 150 μl of 1 μM of MM12ap intravenously 4, 6 and 8 weeks after DMM or sham surgery. Images were taken 2, 4 and 8 h after injection and used to quantify the MFI in the operated and contralateral knees. Whiskers depict the 95% CI. n = 6 for each group. Analysis by two-way ANOVA with Bonferroni post-tests indicated no significant difference between any of the groups at any time points between the weeks. Osteoarthritis and Cartilage 2014 22, 862-868DOI: (10.1016/j.joca.2014.04.006) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Quantification of the activation of MMP13ap in the DMM model of OA mice were injected with 150 μl of 1 μM of the MMP13ap intravenously 4, 6 and 8 weeks after DMM or sham surgery. Images were taken 2, 4 and 8 h after injection and used to quantify the MFI in the operated and contralateral knees (A). Whiskers depict the 95% ci. n = 6 for each group in week 4, n = 9 for each group in week 6 and n = 8 for each group in week 8. *P < 0.05 between DMM and other groups after two-way ANOVA analysis with Bonferroni post tests (DMM vs contra: P = 0.0477, DMM vs Sham: P = 0.0421, DMM vs Sham contra: P = 0.0161). (B) The difference in signal between the operated and contralateral knees in the DMM and sham groups. *P < 0.05 by paired Student's t test, individual values as indicated. Osteoarthritis and Cartilage 2014 22, 862-868DOI: (10.1016/j.joca.2014.04.006) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Longitudinal correlation of the signal from the MMP13ap in the progression of the DMM model of OA The MFI of the relevant ROIs obtained at 4 h after injection of the MMP13ap was plotted against either the duration after surgery (A) or the individual summed histological scores 4 weeks (B), 6 weeks (C) or 8 weeks (D) after surgery. *P < 0.05 between DMM and other groups, as detailed in Fig. 4. The Pearson correlation factor, r at P < 0.005, between mean fluorescent signal and histological damage for weeks 6 and 8 are indicated. Osteoarthritis and Cartilage 2014 22, 862-868DOI: (10.1016/j.joca.2014.04.006) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions