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Abstract

Objectives

To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone.

Animals

2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2).

Procedure

Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(–) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies.

Results

Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(–) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(–) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization.

Conclusions

Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(–) epitope immunoreactivity and chondrocyte hypertrophy. (Am J Vet Res 1996;57:1080–1093)

Free access
in American Journal of Veterinary Research

Summary

Intra-articularly administered, long-acting corticosteroids are a beneficial treatment for many equine joint disorders because they alleviate inflammation and signs of pain, but they also exert detrimental effects on the biochemical composition and morphologic features of articular cartilage. Chondroprotective drugs have been shown to mitigate some of the deleterious effects of intra-articularly administered corticosteroids on articular cartilage of laboratory animals. Twenty-one ponies were assigned at random to receive 1 of 3 treatments in the right middle carpal joint. Group-1 ponies (n = 8) had methylprednisolone acetate (mpa; 0.2 mg/kg of body weight) and saline solution administered intra-articularly and im, respectively. Group-2 ponies (n = 9) received mpa (0.2 mg/kg) and polysulfated glycosaminoglycan (gag; 2 mg/kg). Group-3 ponies (control; n = 4) had saline solution administered intra-articularly and im. The corticosteroid or saline solution was injected into the right middle carpal joint on day 1. The im administered polysulfated gag or saline solution was administered at the same time, then was repeated every 3 days for 20 days. Ponies were euthanatized 21 days after initial injection by overdose of pentobarbital sodium.

The cartilage of younger ponies was significantly (P < 0.05) more responsive to the proteoglycan-depleting effects of mpa. Ponies < 10 years old of groups 1 and 2 had significantly (P < 0.05) lower gag content in the articular cartilage than did control ponies. Systemic treatment with polysulfated gag did not result in a protective effect against proteoglycan loss from the articular cartilage. Twenty-one days after mpa injection, difference in [35S]sulfate incorporation into proteoglycan, between either mpa-treated group and the control group, was not significant. There was an approximate tenfold increase in keratan sulfate concentration in synovial fluid from mpa-treated joints, compared with control joints. Chondroprotective effect of polysulfated gag on the basis of keratan sulfate release from the articular cartilage into the synovial fluid was not observed. Methylprednisolone acetate caused a decrease in the fibronectin content of articular cartilage, but there was no effect of polysulfated gag on the fibronectin content of mpa-treated articular cartilage.

Free access
in American Journal of Veterinary Research