Differential transcriptome analysis of intraarticular lesional vs intact cartilage reveals new candidate genes in osteoarthritis pathophysiology M. Geyer,

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Presentation transcript:

Differential transcriptome analysis of intraarticular lesional vs intact cartilage reveals new candidate genes in osteoarthritis pathophysiology M. Geyer, M.D., S. Grässel, Ph.D., R.H. Straub, M.D., G. Schett, M.D., R. Dinser, M.D., J. Grifka, M.D., S. Gay, M.D., E. Neumann, Ph.D., U. Müller-Ladner, M.D. Osteoarthritis and Cartilage Volume 17, Issue 3, Pages 328-335 (March 2009) DOI: 10.1016/j.joca.2008.07.010 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Swing mill (microdismembrator) used for cartilage preparation (A). The individual cartilage tissue samples were pulverized with steels balls in steel cases (B) double-fixed in the mill (A) with a safety screw. The pre- (C) and post-pulverization (D) status is shown schematically. Osteoarthritis and Cartilage 2009 17, 328-335DOI: (10.1016/j.joca.2008.07.010) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Graphical analysis of the RNA integrity on the Agilent 2100 Bioanalyzer. The electropherogram shows clear peaks for both 28S and 18S rRNA. Although there is a slight decrease in height of the 28S rRNA peak, RNA quality can be considered as high. Osteoarthritis and Cartilage 2009 17, 328-335DOI: (10.1016/j.joca.2008.07.010) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Representative results of a quantitative RT-PCR. Intensity of differential expression is indicated as the logarithm base 2 of the fold change. RQ: relative quantification, equal to fold change. Osteoarthritis and Cartilage 2009 17, 328-335DOI: (10.1016/j.joca.2008.07.010) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Illustration of representative cartilage sections within the same joint of OA patients (first panel: original magnification 100×; subsequent panels: original magnification 200×). The first panel shows an intact area (left, A) and a diseased area (right, B) stained by hematoxylin–eosin. The second panel shows the lack of expression of AQP-1 in non-lesional areas of an OA patient (C, phase contrast D), whereas the majority of chondrocytes in the superficial zone of an OA lesion express intensively AQP-1 (↑, E, phase contrast F). Similarly, WISP-1 is expressed throughout the areas of cartilage erosions (↑, I and K), whereas in intact cartilage, no specific staining (G and H) can be detected. This is also visible for DAF. Here, an increased staining is to be observed in lesional areas (↑, N and O) in contrast to unaffected cartilage (L and M). Note that for WISP-1 (third panel, H and K) and DAF (fourth panel, M and O) counterstaining with Mayer's hematoxylin was performed to detect the nuclei of the chondrocytes. The inserts in K and O show a larger image of single counter-stained chondrocytes expressing WISP-1 (K) and DAF (O), respectively. Black and white scale bars always indicate 100μm. Osteoarthritis and Cartilage 2009 17, 328-335DOI: (10.1016/j.joca.2008.07.010) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions