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Equine veterinary journal2017; 49(5); 662-667; doi: 10.1111/evj.12666

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

Abstract: Clinical tools to diagnose the early changes of osteoarthritis (OA) that occur in the articular cartilage are lacking. Objective: We 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. Methods: In vitro quantitative proteomics and assay development with application in synovial fluids samples obtained from biobanks of well-characterised horses. Methods: Articular cartilage explants were incubated with or without interleukin-1β 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. Results: Semitryptic 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-1β-stimulated explants. Conclusions: The ELISA is based on polyclonal antisera rather than a monoclonal antibody. Conclusions: The 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.
Publication Date: 2017-02-28 PubMed ID: 28097685PubMed Central: PMC5573946DOI: 10.1111/evj.12666Google Scholar: Lookup
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  • Journal Article

Summary

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The research article presents the identification of a new biomarker, named cartilage oligomeric matrix protein (COMP) neoepitope, that has potential for early osteoarthritis diagnosis in horses. The COMP neoepitope was found to be present in higher concentrations in the synovial fluid of horses experiencing acute lameness, a possible early sign of osteoarthritis, compared to healthy horses or horses with chronic lameness.

Methods and Materials

  • The study employed quantitative proteomics and assay development, with a focus on synovial fluid samples from a well-catalogued horse biobank. The samples were derived from healthy horses, horses with varying stages of osteoarthritis, and horses causing lameness.
  • Articular cartilage explants were cultivated in the presence or absence of interleukin-1β – a cytokine known to influence osteoarthritis development – for 25 days, and analyzed using quantitative proteomics.
  • An inhibition enzyme-linked immunosorbent assay (ELISA) designed for this study was utilized to measure the concentration of the COMP neoepitope in the synovial fluid samples.
  • The explants were immunostained using antibodies against COMP and the COMP neoepitopes for further analysis.

Results and Discussion

  • Semitryptic COMP peptides were identified and quantified from the cell culture media obtained from the cartilage explants.
  • An antibody against the COMP neoepitope was produced and was successful in detecting the COMP neoepitope in the synovial fluid through the custom-developed inhibition ELISA.
  • Higher concentration of the COMP neoepitope was found in the synovial fluid from joints causing acute lameness than in those from normal joints or chronic lameness causing joints.
  • The COMP neoepitope showed a pericellular presence in the explants stimulated with interleukin-1β.
  • It’s important to note that the ELISA test was based on the polyclonal antisera rather than a monoclonal antibody – a limitation that may influence the results.

Conclusions

  • The increase of the COMP neoepitope in the synovial fluid of horses causing acute lameness suggests this neoepitope may potentially serve as a novel biomarker for early molecular changes in articular cartilage connected to osteoarthritis.
  • This new diagnostic tool could assist in providing early and accurate osteoarthritis diagnosis, leading to a more effective and timely treatment of the disease.

Cite This Article

APA
Skiöldebrand E, Ekman S, Mattsson Hultén L, Svala E, Björkman K, Lindahl A, Lundqvist A, Önnerfjord P, Sihlbom C, Rüetschi U. (2017). Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis. Equine Vet J, 49(5), 662-667. https://doi.org/10.1111/evj.12666

Publication

ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 49
Issue: 5
Pages: 662-667

Researcher Affiliations

Skiöldebrand, E
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
  • Division of Pathology, Pharmacology and Toxicology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Ekman, S
  • Division of Pathology, Pharmacology and Toxicology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Mattsson Hultén, L
  • Department of Clinical Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden.
Svala, E
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
  • Division of Pathology, Pharmacology and Toxicology, Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Björkman, K
  • Department of Clinical Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden.
Lindahl, A
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.
Lundqvist, A
  • Department of Clinical Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden.
Önnerfjord, P
  • Section for Rheumatology and Molecular Skeletal Biology, Department of Clinical Sciences Lund, Lund University, Lund, Sweden.
Sihlbom, C
  • Proteomics Core Facility, Sahlgrenska Academy, Gothenburg University, Gothenburg, Sweden.
Rüetschi, U
  • Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Gothenburg University, Gothenburg, Sweden.

MeSH Terms

  • Animals
  • Biomarkers
  • Cartilage Oligomeric Matrix Protein / cerebrospinal fluid
  • Glycoproteins
  • Horse Diseases / cerebrospinal fluid
  • Horses
  • Lameness, Animal / cerebrospinal fluid
  • Matrilin Proteins
  • Osteoarthritis / cerebrospinal fluid
  • Osteoarthritis / veterinary
  • Synovial Fluid / metabolism

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Citations

This article has been cited 13 times.
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