Elevated synovial fluid levels of interleukin-6 and tumor necrosis factor associated with early experimental canine osteoarthritis.
OBJECTIVE
To measure levels of cytokines, proteases, and glycosaminoglycans (GAG) in synovial fluid (SF) from the knees of animals with experimental osteoarthritis (OA) and from their contralateral (control) knees, and to compare and correlate these values with each other as well as with measures of proteoglycan synthesis in the corresponding articular cartilage. This study will help to identify cytokines of potential importance in the early stages of the development of OA.


METHODS
OA was induced in 12 mature animals by sectioning the anterior cruciate ligament. After 3 months, SF from the operated and contralateral (control) knee joints was assayed for interleukin-6 (IL-6), tumor necrosis factor (TNF), IL-1, latent metalloproteinase, and sulfated GAG. Proteoglycan synthesis in the corresponding articular cartilage was also measured.


RESULTS
IL-6 levels in SF from the operated joint compared with the control joint were significantly elevated in 11 of 12 animals. TNF levels were also elevated in 10 of 11 SF samples from operated joints, but to a lesser extent than those of IL-6. IL-1 and IL-1 inhibitors were undetectable in either the operated or control joint SF. The GAG concentration was elevated in SF from experimental OA joints. This elevation correlated with that of TNF, but not IL-6. There was no significant difference in the concentration of APMA-activatable metalloproteinase. The rate of proteoglycan synthesis was higher in the cartilage from the operated joint in 8 of 12 animals, and the mean rate of synthesis was significantly higher than in the control joint. There was a positive correlation between this increase in cartilage proteoglycan synthesis (operated versus control) and the increase in SF IL-6, but there was no correlation with the levels of TNF or GAG.


CONCLUSIONS
This is the first study of SF levels of cytokines in early experimental OA. Our results show surprisingly high levels of IL-6 in operated joints, where the cytokine could act directly on the chondrocytes, and thus play a role in mediating their responses to cartilage injury.
Biochemistry Division, Kennedy Institute of Rheumatology, Hammersmith, London, United Kingdom.
G
G
Venn
J J
JJ
Nietfeld
A J
AJ
Duits
F M
FM
Brennan
E
E
Arner
M
M
Covington
M E
ME
Billingham
T E
TE
Hardingham
D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism; D004195:Disease Models, Animal; D004285:Dogs; D005260:Female; D006025:Glycosaminoglycans / Q000378:metabolism; D007375:Interleukin-1 / Q000032:analysis*; D015850:Interleukin-6 / Q000032:analysis*; D007719:Knee Joint; D008666:Metalloendopeptidases / Q000032:analysis; D010003:Osteoarthritis / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013582:Synovial Fluid / Q000737:chemistry* / Q000201:enzymology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D006025:Glycosaminoglycans; D007375:Interleukin-1; D015850:Interleukin-6; D011509:Proteoglycans; D013431:Sulfates; D013462:Sulfur Radioisotopes; D014409:Tumor Necrosis Factor-alpha; D008666:Metalloendopeptidases