Title page for ETD etd-07112005-184347

Type of Document Master's Thesis
Author Burkert, Blaine Allen
URN etd-07112005-184347
Title Matrix Metalloproteinase 3, Matrix Metalloproteinase 13, and Tissue Inhibitor of Metalloproteinase 1 Concentrations in Normal and Naturally-Occurring Osteoarthritic Canine Stifles
Degree Master of Science (M.S.)
Department Veterinary Clinical Sciences (Veterinary Medical Sciences)
Advisory Committee
Advisor Name Title
Giselle L. Hosgood Committee Chair
Glenna E. Mauldin Committee Member
Loretta J. Bubenik Committee Member
  • cranial cruciate ligament rupture
  • matrix metalloproteinase
  • osteoarthritis
Date of Defense 2005-06-21
Availability unrestricted
Osteoarthritis is arguably the most common ailment in both dogs and people in the developed World. Treatment for osteoarthritis is currently symptomatic. Development of therapies designed at stopping the progression of osteoarthritis, require a method of evaluating efficacy. Objective analysis by measuring joint metabolism via clinical trials of the cross-over design is currently not possible with the methods utilized.

Extracellular matrix degradation is a hallmark of osteoarthritis. The matrix metalloproteinases are major degradative enzymes. There are currently no commercially available assays to measure canine matrix metalloproteinases. Human and canine matrix metalloproteinases are highly homologous. The use of antibodies and assays designed to detect human matrix metalloproteinases may also detect canine molecules.

A commercially available human enzyme-linked immunosorbent assay (ELISA) was tested against canine samples without success.

Casein zymography detected the presence of canine enzymes weighing the same as human matrix metalloproteinase 3 (MMP-3) and matrix metalloproteinase 13 (MMP-13). Inhibition of the casein degradation was achieved by the addition of ethylenediamine tetra-acetic acid (EDTA). The presence of a protein of the correct molecular weight with the ability to digest casein that is inhibited by EDTA is only circumstantial and does not definitively identify the enzymes.

Enzyme-assisted immunoelectroblotting (Western blotting) utilizing human polyclonal antibodies was unsuccessful at positively identifying the canine molecule.

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