M. Stock, A. Distler, J. Distler, C. Beyer, G. Ruiz-Heiland, N

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Fc-gamma receptors are not involved in cartilage damage during experimental osteoarthritis M. Stock, A. Distler, J. Distler, C. Beyer, G. Ruiz-Heiland, N. Ipseiz, M. Seeling, G. Krönke, F. Nimmerjahn, G. Schett Osteoarthritis and Cartilage Volume 23, Issue 7, Pages 1221-1225 (July 2015) DOI: 10.1016/j.joca.2015.02.019 Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 FcγRs are expressed in murine chondrocytes. A: Total RNA was extracted from murine chondrocytes, newborn epiphyseal cartilage, adult (8 weeks) articular cartilage and macrophages. RT-PCR analyses were performed to detect mRNA levels of FcγR types I, IIB, III, and IV, collagen2a1 (Col2a1) and CD45. Cyclophilin A was used as a reference gene. ECart: epiphyseal cartilage; EC: epiphyseal chondrocytes; RC rib chondrocytes; ACart: articular cartilage; MΦ (bm): bone marrow macrophages. B: Rib chondrocytes from WT or FcγRIIB-/FcRγ-chain-double-deficient (KO) mice were analysed for FcγR cell surface expression using flow cytometry. Shown is the delta median fluorescence intensity (Δ MFI) compared with unstained samples of a typical experiment. All experiments shown were performed at least three times with comparable results. Osteoarthritis and Cartilage 2015 23, 1221-1225DOI: (10.1016/j.joca.2015.02.019) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Deficiency in FcγRs does not alter cartilage damage in DMM-induced OA. DMM or sham surgery was performed in WT, FcγRIIB-deficient (RIIB), or FcR γ-chain-deficient (γ) mice. Eight weeks post-surgery, cartilage damage and osteophyte development was investigated histologically. A: Safranin-O staining of frontal knee sections showing the medial part of the stifle joint. B: Quantification of proteoglycan (PG) loss, determined as the fraction of Safranin-O staining negative articular cartilage (Saf-O neg. cart.). C: Scoring of OA-induced cartilage damage according to OARSI recommendations. Maximal scorings of all four quadrants of the joint were cumulated per mouse. D: Osteophyte development was determined by measuring the size of osteophytes at the medial tibiae. Mean values ± 95% CI are shown. n = 6. Osteoarthritis and Cartilage 2015 23, 1221-1225DOI: (10.1016/j.joca.2015.02.019) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions