Article

Viability and cell death of synovial fluid neutrophils as diagnostic biomarkers in equine infectious joint disease: a pilot study.

Department of Pharmacology, Toxicology and Biochemistry, Ghent University, Belgium.
Research in Veterinary Science (impact factor: 1.65). 11/2010; 92(1):132-7. DOI:10.1016/j.rvsc.2010.10.007 pp.132-7
Source: PubMed

ABSTRACT Synovial fluid samples from culture-confirmed infected joints (n=13), joints with pronounced non-infectious synovitis (n=11) and healthy joints (n=14) were collected from 24 equine patients and seven slaughterhouse horses. The samples from the joints with non-infectious synovitis and healthy joints served as negative controls. After isolation, counting and identification of neutrophils, the percentage viability, and the proportion apoptotic and necrotic neutrophils were determined by flow cytometry. Viability was significantly higher in infected samples compared to the controls. A significant difference in cell death type was observed, with apoptosis predominating in infected joints, and necrosis being more present in joints with pronounced non-infectious synovitis and healthy joints. The results of this pilot study suggest that flow cytometric analysis of neutrophil viability and cell death dynamics may assist the discrimination between infected and non-infected joints.

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    Article: Equine myeloperoxidase: A novel biomarker in synovial fluid for the diagnosis of infection.
    J Wauters, F Pille, A Martens, T Franck, D Serteyn, F Gasthuys, E Meyer
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    ABSTRACT: REASONS FOR PERFORMING STUDY: Equine joint infection is a life-threatening disorder, and confirmation of the diagnosis can be difficult. Synovial fluid biomarkers may assist the discrimination between infectious and noninfectious joint disease. OBJECTIVES: This study investigates whether the immunological detection of total and enzymatically active myeloperoxidase (MPO) assists the diagnosis of joint infection in horses. METHODS: The following 4 sample groups were included: healthy; osteochondritis dissecans (OCD); traumatic synovitis; and culture-confirmed infected joints. Synovial fluid was analysed for total MPO by a horse-specific sandwich enzyme-linked immunosorbent assay (ELISA) and for active MPO using the specific immunological extraction followed by enzymatic detection (SIEFED) technique. Western blot analysis was performed to confirm the antibody specificity. RESULTS: Synovial fluid from infected joints contained significantly more total and active MPO than samples from healthy joints, joints affected by OCD and joints with traumatic synovitis. Cut-off values were set at 5000 and 350 ng/ml for total and active MPO, respectively, with fair sensitivity, specificity, positive and negative predictive values and likelihood ratios for infection. Correlation coefficients were reported between the total as well as the active MPO levels and the routine synovial fluid parameters, i.e. the white blood cell count, the neutrophil count and the total protein level. No correlation was observed between MPO and either the age of the horse or the joint affected. Western blotting confirmed the antibody specificity for equine MPO. CONCLUSIONS AND POTENTIAL RELEVANCE: Synovial fluid MPO was identified as a very promising biomarker to augment the discrimination of infectious vs. noninfectious joint disease in horses. Both ELISA and SIEFED techniques can be used for its specific and rapid detection. The analysis of synovial fluid MPO can be used as a complementary test to aid in the discrimination between infectious and noninfectious joint disease, especially when the white blood cell counts and the total protein level are inconclusive.
    Equine Veterinary Journal 09/2012; · 1.46 Impact Factor
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Keywords

24 equine patients
 
apoptosis predominating
 
cell death dynamics
 
cell death type
 
flow cytometric analysis
 
flow cytometry
 
healthy joints
 
joints
 
negative controls
 
neutrophil viability
 
non-infected joints
 
non-infectious synovitis
 
percentage viability
 
proportion apoptotic
 
Synovial fluid samples
 
Viability