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Synovial fluid gelatinase concentrations and matrix metalloproteinase and cytokine expression in naturally occurring joint disease in horses

Troy N. TrumbleEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Gayle W. TrotterEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Julie R. Thom OxfordEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.
Present address is Oregon Health Sciences University, Portland, OR 97201.

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C. Wayne McIlwraithEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Sheryl CammarataEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.
Present address is PO Box 344, Masonville, CO 80541.

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Jennifer L. GoodnightEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.
Present address is Cardiovascular Institute, Department of Cardiology, Aurora, CO 80010.

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R. Clark BillinghurstEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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David D. FrisbieEquine Orthopaedic Research Laboratory and the Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Abstract

Objectives—To determine concentrations of matrix metalloproteinase (MMP)-2 and -9 in synovial fluid; and mRNA expression of MMP-1, -13, and -3; interleukin[ IL]-1α and β; and tumor necrosis factor(TNF)-α in synovial membrane and articular cartilage from horses with naturally occurring joint disease.

Sample Population—Synovial fluid (n = 76), synovial membrane (59), and articular cartilage (45) from 5 clinically normal horses and 55 horses with joint disease categorized as traumatic (acute [AT] or chronic [CT]), osteochondritis dissecans (OCD), or septic (S).

Procedure—Synovial fluid gelatinase concentrations were analyzed, using zymography. Synovial membrane and articular cartilage mRNA expression for MMP-1, -3, and -13, IL-1α and β, TNF-α, type-II collagen, and aggrecan were analyzed, using quantitative reverse transcriptase-polymerase chain reaction.

Results—Synovial fluid pro-MMP-2 concentration was significantly higher in diseased joints than normal joints. Septic joints had significantly higher concentrations of pro and active MMP-9. Stromelysin-1 was expressed in ≥ 80% of synovial membrane and articular cartilage samples and was strongly influenced by age. Collagenases were rarely expressed, with MMP- 13 expressed only in diseased joints. Interleukin-1β expression was significantly higher in all OCD samples and was influenced by age. Tumor necrosis factor- α expression was significantly higher in cartilage from joints with AT and OCD. There was no correlation between MMP or cytokines and type-II collagen or aggrecan expression.

Conclusions and Clinical Relevance—Matrix metalloproteinase- 2 and -3 are abundant in naturally occurring joint disease and normal joints. Interleukin-1β and TNF-α may be important in the pathogenesis of OCD. Age affects MMP and IL-1β concentrations. (Am J Vet Res 2001;62:1467–1477)

Abstract

Objectives—To determine concentrations of matrix metalloproteinase (MMP)-2 and -9 in synovial fluid; and mRNA expression of MMP-1, -13, and -3; interleukin[ IL]-1α and β; and tumor necrosis factor(TNF)-α in synovial membrane and articular cartilage from horses with naturally occurring joint disease.

Sample Population—Synovial fluid (n = 76), synovial membrane (59), and articular cartilage (45) from 5 clinically normal horses and 55 horses with joint disease categorized as traumatic (acute [AT] or chronic [CT]), osteochondritis dissecans (OCD), or septic (S).

Procedure—Synovial fluid gelatinase concentrations were analyzed, using zymography. Synovial membrane and articular cartilage mRNA expression for MMP-1, -3, and -13, IL-1α and β, TNF-α, type-II collagen, and aggrecan were analyzed, using quantitative reverse transcriptase-polymerase chain reaction.

Results—Synovial fluid pro-MMP-2 concentration was significantly higher in diseased joints than normal joints. Septic joints had significantly higher concentrations of pro and active MMP-9. Stromelysin-1 was expressed in ≥ 80% of synovial membrane and articular cartilage samples and was strongly influenced by age. Collagenases were rarely expressed, with MMP- 13 expressed only in diseased joints. Interleukin-1β expression was significantly higher in all OCD samples and was influenced by age. Tumor necrosis factor- α expression was significantly higher in cartilage from joints with AT and OCD. There was no correlation between MMP or cytokines and type-II collagen or aggrecan expression.

Conclusions and Clinical Relevance—Matrix metalloproteinase- 2 and -3 are abundant in naturally occurring joint disease and normal joints. Interleukin-1β and TNF-α may be important in the pathogenesis of OCD. Age affects MMP and IL-1β concentrations. (Am J Vet Res 2001;62:1467–1477)