Determination of MMP-2 and -9 activities in synovial fluid of horses with osteoarthritic and arthritic joint diseases using gelatin zymography and immunocapture activity assays

Equine Vet J. 2008 May;40(3):266-71. doi: 10.2746/042516408X251921.

Abstract

Reason for performing study: Matrix metalloproteinases (MMPs)-2 and -9 activities have been found elevated in synovial fluid from various joint diseases in man. However, in the horse few data are available.

Objectives: To explore the clinical significance of MMP-2 and -9 activities in synovial fluid of horses with different forms of joint diseases.

Methods: Gelatin zymography and MMP-2 and -9 immunocapture activity assays were applied on synovial fluids from control joints and joints with aseptic joint disease (AJD) and septic arthritis (SA). Additionally, MMP-2 and -9 activities were measured in samples from SA to monitor the disease process.

Results: Zymographic analysis revealed that samples from AJD and SA contained significantly increased latent MMP-2 activity compared to controls. Samples from SA showed significantly increased monomeric latent MMP-9 activity compared with all other affected joints and controls. Trace amounts of MMP-9 activity, due to the active and dimer form, were detected in samples from SA; however, these bands were absent in samples from AJD and controls. Using immunocapture activity assays, MMP-2 and -9 activities were found to be significantly elevated in joints from SA compared to controls and AJD samples. MMP-2 activity in samples from AJD was significantly increased compared to controls. Both MMP activities decreased in the joints from SA in the course of successful therapy.

Conclusions: Data from zymographic analysis confirmed that MMP-2 and -9 were elevated in equine joint diseases. Immunocapture activity assays have been shown to be suitable for the quantitative determination of MMP-2 and -9 activities in synovial fluid of horses. Both MMP-2 and -9 activities seem to be useful to indicate SA, and MMP-2 activity might be a suitable marker for AJD.

Potential relevance: These findings encourage the potential use of MMP-2 and -9 as additional aids to clinical investigation. Further work is required to validate the clinical significance of MMP activities in the progress of different joint diseases in horses.

MeSH terms

  • Animals
  • Biomarkers / analysis
  • Biomarkers / metabolism
  • Disease Progression
  • Horse Diseases / diagnosis
  • Horse Diseases / enzymology*
  • Horses
  • Joint Diseases / diagnosis
  • Joint Diseases / enzymology
  • Joint Diseases / veterinary*
  • Matrix Metalloproteinase 2 / analysis
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 9 / analysis
  • Matrix Metalloproteinase 9 / metabolism*
  • Osteoarthritis / diagnosis
  • Osteoarthritis / enzymology
  • Osteoarthritis / veterinary*
  • Synovial Fluid / enzymology*

Substances

  • Biomarkers
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9