J Vet Sci.  2011 Sep;12(3):273-280. 10.4142/jvs.2011.12.3.273.

Biomarkers for identifying the early phases of osteoarthritis secondary to medial patellar luxation in dogs

• Alam MR
• Ji JR
• Kim MS
• Kim NS

• KMID: 1067400
• DOI: http://doi.org/10.4142/jvs.2011.12.3.273

Abstract

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.

Keyword

MMP-2; osteoarthritis; synovial fluid; TIMP-2; TRAP

MeSH Terms

Acid Phosphatase/analysis/blood
Animals
Arthritis, Experimental/enzymology/etiology/veterinary
Biological Markers/*analysis/*blood
Blotting, Western/veterinary
Dislocations/complications/*veterinary
Dog Diseases/*enzymology/etiology
Dogs
Enzyme-Linked Immunosorbent Assay/veterinary
Female
Isoenzymes/analysis/blood
Male
Matrix Metalloproteinase 2/analysis/blood
Osteoarthritis/enzymology/etiology/*veterinary
Spectrophotometry/veterinary
Stifle/physiopathology
Synovial Fluid/*enzymology
Tissue Inhibitor of Metalloproteinase-2/analysis/blood

Figure

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  Fig. 1 Tartrate resistant acid phosphatase (TRAP) levels in the synovial fluid (SF) of the index and contralateral (control and sham) stifles after induction of osteoarthritis (OA) during different experimental periods.

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  Fig. 2 TRAP levels in the serum during different experimental periods after inducing OA secondary to medial patellar luxation (MPL).

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  Fig. 3 Tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) levels in the SF of the index and contralateral (control and sham) stifles during different experimental periods after OA induction.

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  Fig. 4 TIMP-2 levels in the serum at different experimental periods after inducing OA secondary to MPL.

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  Fig. 5 Western blot showing matrix metalloproteinase-2 (MMP-2) levels in the SF during different experimental periods after the induction of OA secondary to MPL.

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  Fig. 6 Microphotographs of the stained frozen synovium section. The red mononuclear cells (arrow) are positive for TRAP. (A) Control. (B) OA 3 month. (C) OA 6 month. (D) OA 12 month. Scale bars = 100 µm.

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  Fig. 7 Microphotographs of the stained frozen cranial cruciate ligament (CCL) section. The red mononuclear cells (arrow) are positive for TRAP. (A) Control. (B) OA 3 month. (C) OA 6 month. (D) OA 12 month. Scale bars = 100 µm.

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  Fig. 8 Microphotographs of the stained frozen articular cartilage section. The red mononuclear cells (arrow) are positive for TRAP. (A) Control. (B) OA 12 month. Scale bars = 100 µm.

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