Comparison of periprosthetic tissues in knee and hip joints: differential expression of CCL3 and DC-STAMP in total knee and hip arthroplasty and similar.

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Comparison of periprosthetic tissues in knee and hip joints: differential expression of CCL3 and DC-STAMP in total knee and hip arthroplasty and similar cytokine profiles in primary knee and hip osteoarthritis T. Tomankova, E. Kriegova, R. Fillerova, P. Luzna, J. Ehrmann, J. Gallo Osteoarthritis and Cartilage Volume 22, Issue 11, Pages 1851-1860 (November 2014) DOI: 10.1016/j.joca.2014.08.004 Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Distribution of relative mRNA expression (ratio target gene/reference HPRT1 gene) of (A) TNF, (B) IL-6, (C) IL-8, (D) BMP4, (E) CHIT1, (F) MMP9, (G) CCL18, (H) CCL3, (I) DC-STAMP, (J) RANKL, (K) OPG, and (L) SOCS3 in pseudosynovial membrane tissues obtained from patients with aseptically failed TKA and THA and in synovial membrane obtained from patients with primary knee (OA-knee) and hip (OA-hip) OA. Group means are indicated by horizontal bars, error bars indicate 95% CI; only P-values for significant differences between two groups of patients are stated. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Distribution of relative mRNA expression (ratio target gene/reference HPRT1 gene) of CCL3, DC-STAMP, and NF-κB in pseudosynovial membrane tissues obtained from patients with (1) aseptically failed TKA and THA (OL/AL) and (2) in patients with C-TKA and C-THA re-operated for other reasons (pain, instability, fracture), exhibiting no sign of periprosthetic osteolysis/aseptic loosening at the time of tissue sampling (no OL/AL). Group means are indicated by horizontal bars, error bars indicate 95% CI; only P-values for significant differences between two groups of patients are stated. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Immunohistochemical staining for CCL3, DC-STAMP, IL-8, SOCS3, OPG, RANKL in pseudosynovial membrane sections obtained from patients with aseptically failed TKA and THA and in synovial membrane sections from patients with primary knee (OA-knee) and hip (OA-hip) OA – representative examples. Typical fibroblasts (solid arrow), endothelial cells (dashed arrow), lymphocytes (dotted arrow), multinucleated giant cells (empty arrow) are marked. Stainings with a non-specific isotype control antibody are shown in right upper segment of each panel. Scale bar, 50 μm. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 H&E and immunohistochemical staining of pseudosynovial membrane sections obtained from patients with aseptically failed TKA and THA – representative examples. H&E: Pseudosynovial membrane from aseptically loosened TKA is characterized by presence of large highly nucleated giant cells (A, C); in THA, smaller giant cells with less nuclei are present (B, D). The giant cells in both TKA and THA are formed by osteoclast-like cells as shown by TRAP staining (E, F). Scale bar: A, B 100 μm; C, D, E, F 50 μm. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. E1 Pseudosynovial tissues retrieved during reoperation of a cementless THA in 52-years old man (a) and cemented TKA in 69-years old woman (b); synovial tissue obtained during primary THA from a 65-years old woman with primary OA (c) – representative examples. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. E2 Hierarchical clustering of gene expression data. The rows represent individual genes, the columns represent individual patient samples (one column per sample) divided into four cohorts (TKA, THA, OA-knee, primary knee OA; OA-hip, primary hip OA). The colour represents the gene expression level (green = low expression, red = high expression); the expression levels are continuously mapped on the colour scale provided at the top of the figure. The analysis was performed with the R statistical software package (http://www.r-project.org/). Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. E3 RANKL/OPG mRNA ratio in pseudosynovial membrane tissues obtained from patients with aseptically failed TKA and THA and in synovial membrane obtained from patients with primary knee (OA-knee) and hip (OA-hip) OA. Group means are indicated by horizontal bars, error bars indicate 95% CI. Osteoarthritis and Cartilage 2014 22, 1851-1860DOI: (10.1016/j.joca.2014.08.004) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions