Osteoarthritis and Cartilage

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D. A. Walsh, F. R. C. P. , Ph. D. , C. S. Bonnet, B. Sc. , E. L

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Osteoarthritis and Cartilage Human C-reactive protein aggravates osteoarthritis development in mice on a high-fat diet A.E. Kozijn, M.T. Tartjiono, S. Ravipati, F. van der Ham, D.A. Barrett, S.C. Mastbergen, N.M. Korthagen, F.P.J.G. Lafeber, A.M. Zuurmond, I. Bobeldijk, H. Weinans, R. Stoop Osteoarthritis and Cartilage DOI: 10.1016/j.joca.2018.09.007 Copyright © 2018 The Authors Terms and Conditions

Fig. 1 HFD feeding aggravates OA progression in hCRP-tg mice. A) Representative coronal sections of the lateral and medial tibiofemoral compartments, stained with Fast-Green/Safranin-O, from hCRP-tg and wild-type littermates fed a HFD. Chow controls were not included in our study, but hCRP-tg males are known to not develop OA on a feeding regime of 38 weeks8. Magnification for all microphotographs was 40×. B) Summed histopathological scores for the tibiofemoral knee compartments of the hCRP-tg and wild-type groups. C) Total osteophyte scores showing the individual summed score for the tibiofemoral knee compartments for each animal per study group. D) Total synovitis scores showing the individual summed score for the tibiofemoral knee compartments for each animal per study group. B–D) Scoring was performed according to OARSI histopathology recommendations for the mouse22. Data are presented as group medians (indicated by bars) with IQR (error bars). Osteoarthritis and Cartilage DOI: (10.1016/j.joca.2018.09.007) Copyright © 2018 The Authors Terms and Conditions

Fig. 2 Transgenic and wild-type hCRP males show similar plasma lipid profiles on a HFD. A) Representative PC/LPC pairs show increases in fasted plasma levels in both study groups during the HFD regimen. B) Representative oxylipins from the three most important oxylipin substrates (i.e., AA, LA and DHA) demonstrate distinct concentration changes in fasted plasma during the HFD regimen. Boxplots show boxes extending from the 25th to 75th percentiles containing the median (middle line), with error bars down to the minimum and up to the maximum value. The individual value for each animal is plotted as a dot superimposed on the graph. IS, internal standard; PC, phosphatidylcholine; LPC, lysophosphatidylcholine; EET, epoxyeicosatrienoic acid; DHET, dihydroxyeicosatrienoic acid; HODE, hydroxyoctadecadienoic acid; HDHA, hydroxy docosahexaenoic acid; AA, arachidonic acid; LA, linoleic acid; DHA, docosahexaenoic acid. Osteoarthritis and Cartilage DOI: (10.1016/j.joca.2018.09.007) Copyright © 2018 The Authors Terms and Conditions

Fig. 3 Increased activation of classical and non-classical monocytes in hCRP-tg mice. A) Distribution of peripheral CD11b+ immune cells shows HFD-mediated monocytosis in both groups over time. Baseline population frequencies were determined in age-matched hCRP-tg (n = 5) and wild-type (n = 4) males from later litters and were not included in the statistical analysis (shown here in grey). B) GR-1low non-classical monocytes showed an increase in CCR2 and CD11c expression upon 2 weeks of HFD feeding, which was significantly elevated in hCRP-tg compared with wild-type mice. C-D) GR-1low non-classical monocytes expressing CCR2 upregulated the lymphocyte activation antigen CD86 after long-term HFD feeding (37 weeks), with hCRP-tg males showing higher co-expression of these activation markers compared with wild-type controls. E) GR-1high classical monocytes co-expressing CCR2 and CD86 increase over time and were significantly more present in hCRP-tg compared with wild-type males at t = 37 weeks. Data are presented as group means (indicated by bars) with SD (error bars). *P < 0.05 compared with wild-type controls. Osteoarthritis and Cartilage DOI: (10.1016/j.joca.2018.09.007) Copyright © 2018 The Authors Terms and Conditions

Fig. 4 Human CRP has no effect on synovial macrophage accumulation and subsets. A) Representative unmixed spectral images of the lateral patellofemoral synovial lining (ROI) showing pan-macrophage marker F4/80 (green), M1 subset marker iNOS (blue) and M2 subset marker CD206 (magenta) positivity. Magnification for the top microphotographs was 40×. B) Percentage of the surface area within the ROI or within the F4/80+ cell fraction for hCRP-tg and wild-type mice, showing no CRP-induced shift in macrophage numbers and phenotypes. Data are presented as group medians (indicated by bars) with IQR (error bars). Osteoarthritis and Cartilage DOI: (10.1016/j.joca.2018.09.007) Copyright © 2018 The Authors Terms and Conditions