ABSTRACT
OBJECTIVE: To investigate the effect of adenosine kinase inhibition on interleukin (IL)-1beta- and lipopolysaccharide (LPS)-induced cartilage damage. DESIGN: Articular cartilage was obtained from the metacarpophalangeal joints of 10 young adult horses. Following a stabilization period, weighed cartilage explants were exposed to IL-1beta (10 ng/ml) or LPS (50 microg/ml) to induce cartilage degradation. To test the potential protective effects of adenosine, these explants were simultaneously exposed to adenosine (100 microM), the adenosine kinase inhibitor 5'iodotubercidin (ITU, 1 microM) or to both adenosine and ITU. After 72 h in culture, conditioned medium was collected for evaluation of glycosaminoglycan (GAG), nitric oxide (NO), prostaglandin E2 (PGE2) and matrix metalloproteinase (MMP)-3 release. RESULTS: IL-1beta and LPS stimulated significant release of GAG, NO, PGE2 and MMP-3. Incubation with ITU significantly inhibited both IL-1beta- and LPS-induced GAG release, but did not alter MMP-3 production. Exposure to ITU also reduced IL-1beta-induced PGE2 release and LPS-induced NO production. Direct adenosine supplementation did not attenuate the effects of IL-1beta or LPS, and the addition of adenosine or ITU in the absence of IL-1beta or LPS did not have any detectable effect on cartilage metabolism in this model. CONCLUSIONS: The adenosine kinase inhibitor ITU attenuated experimentally induced cartilage damage in an in vitro cartilage explant model. Release of adenosine from chondrocytes may play a role in the cellular response to tissue damage in arthritic conditions and modulation of these pathways in the joint may have potential for treatment of arthropathies.
Subject(s)
Adenosine Kinase/antagonists & inhibitors , Cartilage, Articular/drug effects , Enzyme Inhibitors/pharmacology , Interleukin-1/toxicity , Lipopolysaccharides/toxicity , Tubercidin/analogs & derivatives , Adenosine/pharmacology , Animals , Cartilage, Articular/metabolism , Culture Media, Conditioned , Dinoprostone/metabolism , Glycosaminoglycans/metabolism , Horses , Interleukin-1/antagonists & inhibitors , Lipopolysaccharides/antagonists & inhibitors , Matrix Metalloproteinase 3/metabolism , Nitric Oxide/metabolism , Tissue Culture Techniques/methods , Tubercidin/pharmacologyABSTRACT
OBJECTIVE: To determine rate and degree of cooling for the superficial digital flexor tendon (SDFT) during a standard cryotherapy application in horses and evaluate in vitro effects of cooling on survival of tendon cells. SAMPLE POPULATION: 6 limbs of 5 adult horses and cultured cells obtained from SDFT of 3 adult horses during necropsy. PROCEDURE: In vivo data were acquired by use of a thermocouple temperature probe inserted into the SDFT of a forelimb of each standing sedated horse. After baseline temperatures were recorded, a commercial compression splint with circulating coolant was placed on each selected limb, which was then exposed to cold treatment for 60 minutes. Temperatures were recorded at 30-second intervals. Mean minimum core temperature was calculated and used to design a protocol for in vitro cold treatment of cells. Specimens were obtained from the SDFT of horses during necropsy; tendon cells were cultured in suspension and exposed to 1-hour of cold treatment that mimicked the in vivo procedure. Viability of cells after cold treatment was compared with viability of cells maintained at body temperature. RESULTS: After 1 hour of cold treatment, SDFT core temperature was reduced by a mean of 21.8 degrees C, reaching a mean minimum temperature of 10 degrees C. Viability did not differ significantly between cold-treated and control cells. CONCLUSION AND CLINICAL RELEVANCE: Results indicated that topical application of cryotherapy significantly reduced core SDFT temperature in standing sedated horses. Temperatures achieved in vivo during cold treatment were not detrimental to the in vitro viability of tendon cells.