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Research article2015Peer reviewed

An inflammatory equine model demonstrates dynamic changes of immune response and cartilage matrix molecule degradation in vitro

Svala, Emilia; Löfgren, Maria; Sihlbom, Carina; Ruetschi, Ulla; Lindahl, Anders; Ekman, Stina; Skiöldebrand, Eva

Abstract

The molecular aspects of inflammation were investigated in equine articular cartilage explants using quantitative proteomics. Articular cartilage explants were stimulated with interleukin (IL)-1 beta in vitro for 25 days, and proteins released into cell culture media were chemically labeled with isobaric mass tags and analyzed by liquid chromatography-tandem mass spectrometry. A total of 127 proteins were identified and quantified in media from explants. IL-1 beta-stimulation resulted in an abundance of proteins related to inflammation, including matrix metalloproteinases, acute phase proteins, complement components and IL-6. Extracellular matrix (ECM) molecules were released at different time points, and fragmentation of aggrecan and cartilage oligomeric matrix protein was observed at days 3 and 6, similar to early-stage OA in vivo. Degradation products of the collagenous network were observed at days 18 and 22, similar to late-stage OA. This model displays a longitudinal quantification of released molecules from the ECM of articular cartilage. Identification of dynamic changes of extracellular matrix molecules in the secretome of equine explants stimulated with IL-1 beta over time may be useful for identifying components released at different time points during the spontaneous OA process.

Keywords

Horse; IL-1 beta; matrix molecules; osteoarthritis; proteomics

Published in

Connective Tissue Research
2015, Volume: 56, number: 4, pages: 315-325
Publisher: INFORMA HEALTHCARE