Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression.

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Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression in human chondrocytes S.L. Dunn, J.M. Wilkinson, A. Crawford, C.L. Le Maitre, R.A.D. Bunning Osteoarthritis and Cartilage Volume 22, Issue 1, Pages 133-144 (January 2014) DOI: 10.1016/j.joca.2013.10.016 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 A–C. The effect of WIN-55 on IL-1β induced MMP-3 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilage and cultured in monolayer. D–F The effect of WIN-55 on IL-1β induced MMP-13 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilage and cultured in monolayer. Each point represents the average of the repeats per patient. IL-1β (10 ng/ml) induced the gene expression of MMP-3 and -13. Following WIN-55 (10 μM) treatment in combination with IL-1β (10 ng/ml) MMP-3 and -13 gene expression was decreased. WIN-55 (10 μM) alone decreased MMP-3 and -13 gene expression. ***P < 0.001, **P < 0.01 compared to DMSO (0.1 %) vehicle control and +++P < 0.001 compared to IL-1β (10 ng/ml) treatment. N = 9 samples for each treatment group, obtained from three separate patient samples for each grade of tissue (0, 2 and 3) tested in triplicate. Osteoarthritis and Cartilage 2014 22, 133-144DOI: (10.1016/j.joca.2013.10.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 A–C. The effect of WIN-55 on IL-1β induced MMP-3 gene expression in human chondrocytes isolated from grade 0, 2 and 3 cartilage and cultured in alginate beads. D–F. The effect of WIN-55 on IL-1β induced MMP-13 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilage and cultured in alginate beads. Each point represents the average of the repeats per patient. IL-1β (10 ng/ml) induced the gene expression of MMP-3 and -13, following WIN-55 (10 μM) treatment in combination with IL-1β (10 ng/ml) MMP-3 and -13 gene expression was decreased or abolished. WIN-55 (10 μM) alone decreased or abolished MMP-3 and -13 gene expression. *P < 0.05, *P < 0.01, ***P < 0.001 compared to DMSO vehicle control and +++P < 0.001 compared to IL-1β treatment. N = 9 samples for each treatment group, obtained from three separate patient samples for each grade of tissue (0, 2 and 3) tested in triplicate. Osteoarthritis and Cartilage 2014 22, 133-144DOI: (10.1016/j.joca.2013.10.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 A–C The effect of WIN-55 on TIMP-1 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilage and cultured in monolayer. D–F. The effect of WIN-55 on TIMP-2 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilages and cultured in monolayer. Each point represents the average of the repeats per patient. IL-1β (10 ng/ml) had no effect on TIMP-1 and -2 gene expression. WIN-55 treatment (10 μM) both alone and in combination with IL-1β (10 ng/ml) reduced TIMP-1 and -2 gene expression. ***P < 0.001 compared to DMSO vehicle control and +++P < 0.001 compared to IL-1β treatment. N = 9 samples for each treatment group, obtained from 3 separate patient samples for each grade of tissue (0, 2 and 3) tested in triplicate. Osteoarthritis and Cartilage 2014 22, 133-144DOI: (10.1016/j.joca.2013.10.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 A–C. The effect of WIN-55 on TIMP-1 gene expression in human OA chondrocytes isolated from grade 0, 2 and 3 cartilages and cultured in alginate beads. D–F. The effect of WIN-55 on TIMP-2 gene expression in OA human chondrocytes isolated from grade 0, 2 and 3 cartilages and cultured in alginate beads. Each point represents the average of the repeats per patient. IL-1β (10 ng/ml) stimulation increased TIMP-1 gene expression and decreased TIMP-2 gene expression below basal levels. WIN-55 treatment (10 μM) both alone and in combination with IL-1β (10 ng/ml) reduced TIMP-1 and -2 gene expression. *P < 0.05, **P < 0.01, ***P < 0.001 compared to DMSO vehicle control and +++P < 0.001 compared to IL-1β treatment. N = 9 samples for each treatment group, obtained from 3 separate patient samples for each grade of tissue (0, 2 and 3) tested in triplicate. Osteoarthritis and Cartilage 2014 22, 133-144DOI: (10.1016/j.joca.2013.10.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Cannabinoid receptor expression in OA chondrocytes cultured in monolayer. Representative photomicrographs of immunocytochemistry of (A) CB1, (B) CB2, (C) PPARα, (D) PPARγ, (E) IgG control (10 μg) (n = 4 cultures). Osteoarthritis and Cartilage 2014 22, 133-144DOI: (10.1016/j.joca.2013.10.016) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions