Detection of collagen type II and proteoglycans in the synovial fluids of patients diagnosed with non-infectious knee joint synovitis indicates early damage to the articular cartilage matrix.
OBJECTIVE
We have sought to determine if markers of proteoglycans and collagen type II (CII) degradation can be detected at an early stage following acute knee injury in the synovial fluid (SF) from a group of patients diagnosed with non-infectious knee joint synovitis (KJS). CII, proteoglycans and elastase activity in the SF from patients with KJS were compared to SF from patients with two chronic arthritis conditions: osteoarthritis (OA) and rheumatoid arthritis (RA) as well as normal SF controls.


METHODS
CII peptides were measured by sandwich ELISA using two monoclonal antibodies: 8:6:D8, a CII-specific antibody, and 14:7:D8 which binds to an amino acid sequence on CII as well as collagens type I, III and V. Epitope 9A4, a neo-epitope resulting from collagenase digestion of CI, CII, and CIII was measured by inhibition ELISA. Proteoglycans measurement included total sulfated glycosaminoglycans (sGAG) by dye-binding assay and 5-D-4 epitope, a keratan sulfate epitope, by inhibition ELISA. Elastase activity was measured colorimetircally using N-succinyl trialanine p-nitroanilide (SANA) substrate.


RESULTS
The quantified CII peptide concentrations by sandwich and inhibition ELISA were significantly higher in SF from patients with KJS (P<0.05) compared to SF from patients with OA, RA and normal aspirates. 5-D-4 and sGAG concentrations were significantly lower (P<0.05) in SF from patients with KJS compared to SF from patients with OA and RA. Elastase activity in SF from patients with KJS and RA were significantly higher (P<0.05) than SF from patients with OA. A significant correlation exists between elastase activity and 9A4 epitope concentration in SF from patients with KJS.


CONCLUSIONS
The elevated CII peptides concentrations in KJS SF compared to normal and OA aspirates indicate early signs of cartilage network damage. The low proteoglycans concentrations in SF from patients with KJS may indicate that injury is limited to the superficial zone of cartilage in the patient population studied. The high elastase activity in SF from patients with KJS and RA are linked to the high CII peptides concentration. The elastase activity in the SF from patients with KJS is due to the action of neutrophil elastase (NE) and collagenases, where both contribute to the destruction of the articular cartilage.
Elsaid
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Department of Biomedical Sciences, University of Rhode Island, Kingston, RI 02881, USA.
Jay
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Chichester
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D001172:Arthritis, Rheumatoid / Q000378:metabolism; D002358:Cartilage, Articular; D024043:Collagen Type II / Q000276:immunology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007632:Keratan Sulfate / Q000276:immunology; D007718:Knee Injuries / Q000150:complications; D007719:Knee Joint / Q000378:metabolism*; D020370:Osteoarthritis, Knee / Q000378:metabolism; D010196:Pancreatic Elastase / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D013582:Synovial Fluid / Q000378:metabolism*; D013585:Synovitis / Q000209:etiology / Q000378:metabolism*
D024043:Collagen Type II; D000939:Epitopes; D011509:Proteoglycans; D007632:Keratan Sulfate; D010196:Pancreatic Elastase