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Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis

Journal article
Authors Eva Skiöldebrand
S. Ekman
L. M. Hulten
Emilia Svala
K. Bjorkman
Anders Lindahl
A. Lundqvist
P. Onnerfjord
Carina Sihlbom
Ulla Ruetschi
Published in Equine Veterinary Journal
Volume 49
Issue 5
Pages 662-667
ISSN 0425-1644
Publication year 2017
Published at Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine
Core Facilities, Proteomics
Pages 662-667
Language en
Links doi.org/10.1111/evj.12666
https://gup.ub.gu.se/file/207111
Keywords horse, cartilage oligomeric matrix protein neoepitope, biomarker, osteoarthritis, synovial fluid, COLLAGEN, DISEASE, BINDING, COMP, INFLAMMATION, DEGRADATION, CLEAVAGE, JOINTS
Subject categories Veterinary Science

Abstract

BackgroundClinical tools to diagnose the early changes of osteoarthritis (OA) that occur in the articular cartilage are lacking. ObjectivesWe sought to identify and quantify a novel cartilage oligomeric matrix protein (COMP) neoepitope in the synovial fluid from the joints of healthy horses and those with different stages of OA. Study designIn vitro quantitative proteomics and assay development with application in synovial fluids samples obtained from biobanks of well-characterised horses. MethodsArticular cartilage explants were incubated with or without interleukin-1 for 25 days. Media were analysed via quantitative proteomics. Synovial fluid was obtained from either normal joints (n = 15) or joints causing lameness (n = 17) or with structural OA lesions (n = 7) and analysed for concentrations of the COMP neoepitope using a custom-developed inhibition enzyme-linked immunosorbent assay (ELISA). Explants were immunostained with polyclonal antibodies against COMP and the COMP neoepitopes. ResultsSemitryptic COMP peptides were identified and quantified in cell culture media from cartilage explants. A rabbit polyclonal antibody was raised against the neoepitope of the N-terminal portion of one COMP fragment (sequence SGPTHEGVC). An inhibition ELISA was developed to quantify the COMP neoepitope in synovial fluid. The mean concentration of the COMP neoepitope significantly increased in the synovial fluid from the joints responsible for acute lameness compared with normal joints and the joints of chronically lame horses and in joints with chronic structural OA. Immunolabelling for the COMP neoepitope revealed a pericellular staining in the interleukin-1-stimulated explants. Main limitationsThe ELISA is based on polyclonal antisera rather than a monoclonal antibody. ConclusionsThe increase in the COMP neoepitope in the synovial fluid from horses with acute lameness suggests that this neoepitope has the potential to be a unique candidate biomarker for the early molecular changes in articular cartilage associated with OA.

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