Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids of patients with temporomandibular joint osteoarthritis.
OBJECTIVE
Imbalance between matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) may be involved in the breakdown of articular cartilage matrix of the temporomandibular joint (TMJ). In this study, MMPs, TIMPs, and MMP-1/TIMP-1 complex levels were examined in TMJ synovial fluid samples aspirated from TMJ osteoarthritis (OA) patients (2 males, 8 females; mean age, 29.7 years) and asymptomatic control subjects (2 males, 8 females; mean age, 23.6 years) to determine the likelihood of increased proteolytic activity in the OA joints.


METHODS
The various types of MMPs and TIMPs were detected by Western blotting with monoclonal antibodies and gelatin zymography. The MMP-1/TIMP-1 complex level was measured by an enzyme-linked immunosorbent assay kit. All aspirates were first analyzed for total protein content and then individually diluted to make the total protein levels equivalent.


RESULTS
The mean MMP-1/TIMP-1 complex concentration in the synovial fluids of the OA patients was 3.92 +/- 1.39 ng/mL; this value was significantly lower (P < 0.05) than the value from control subjects (5.46 +/- 1.32 ng/mL). Matrix metalloproteinase-1 (52 kDa), MMP-3 (57 kDa), TIMP-1 (28 kDa), and TIMP-2 (26 kDa) were detected in all of the normal and the OA samples. However, MMP-1 (28 kDa), MMP-2 (72 kDa), MMP-3 (45 kDa), and MMP-9 (83 kDa) were detected in higher concentration in the OA samples.


CONCLUSIONS
These findings suggest a strong association between the OA-active joints and the presence of biologically active forms of known tissue degradation enzymes (MMP-1, MMP-3, and MMP-9).
Kanyama
M
M
Department of Fixed Prosthodontics, Okayama University Dental School, 2-5-1 Shikata-cho, Okayama 700-8525, Japan.
Kuboki
T
T
Kojima
S
S
Fujisawa
T
T
Hattori
T
T
Takigawa
M
M
Yamashita
A
A
D000293:Adolescent; D000328:Adult; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000201:enzymology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000032:analysis; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D020782:Matrix Metalloproteinases / Q000032:analysis*; D008875:Middle Aged; D010003:Osteoarthritis / Q000201:enzymology* / Q000378:metabolism; D011480:Protease Inhibitors / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D013223:Statistics as Topic; D013582:Synovial Fluid / Q000737:chemistry / Q000201:enzymology*; D013705:Temporomandibular Joint Disorders / Q000201:enzymology* / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000032:analysis; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis*
D000911:Antibodies, Monoclonal; D011480:Protease Inhibitors; D011506:Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; D020782:Matrix Metalloproteinases; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020781:Matrix Metalloproteinase 1