Summary
An easy method of quantitating articular cartilage chondrocyte function in mice is described, using a defined and anatomically intact cartilage structure, the patella. To avoid damage to the cartilage before incubation, 35S incorporation studies were performed leaving the patellae surrounded by a minimal area of non-cartilaginous connective tissue. The patellae were then punched out so that the 35S activity incorporated into the cartilage could be measured. Incorporation of 35S was almost completely blocked by 0.1 mM cycloheximide. Patellar cartilage from left and right knee joints of individual mice shows identical incorporation of 35S, as indicated by a right/left ratio of 1.01 in vitro and 0.99 in vivo, respectively. 35S incorporation values of patellae of mice of different ages do show considerable variation, but within properly age-matched groups of mice the incorporation values are comparable. A known suppressive effect of acute joint inflammation on proteoglycan synthesis was reliably quantitated in the zymosan-induced arthritis model, as indicated by 39% in vitro and 35% in vivo inhibition of 35S incorporation. The patella assay can be used for quantitative measurement of chondrocyte metabolism in normal and pharmacologically altered cartilage.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Mankin HJ, Lippiello L (1969) The turnover of adult rabbit articular cartilage. J Bone Joint Surg [Am] 51:1591–1600
Maroudas A, Evans H (1974) Sulphate diffusion and incorporation into human articular cartilage. Biochim Biophys Acta 338:265–279
Lane JM, Brighton (1974) In vitro rabbit articular cartilage organ model. I. Morphology and glycosaminoglycan metabolism. Arthritis Rheum 17:235–243
Jacoby RK, Jayson MIV (1975) Organ culture of adult human articular cartilage. II. The differential rate of glycosaminoglycan synthesis in layers of articular cartilage matrix. J Rheumatol 2:280–286
Sandy JD, Lowther DA, Brown HLG (1980) Antigen-induced arthritis. Studies on the inhibition of proteoglycan synthesis observed in articular cartilage during short-term joint inflammation. Arthritis Rheum 23:433–447
Mankin HJ, Lippiello L (1970) Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. J Bone Joint Surg [Am] 52:424–434
Mankin HJ (1973) Biochemical and metabolic abnormalities in osteoarthritic human cartilage. Fed Proc 32:1478–1480
Eronen I, Videman T, Friman C, Michelsson J-E (1978) Glycosaminoglycan metabolism in experimental osteoarthrosis caused by immobilization. Acta Orthop Scand 49:329–334
Jacoby RK, Jayson MIV (1976) Synthesis of glycosaminoglycan in adult human cartilage in organ culture from patients with rheumatoid arthritis. Ann Rheum Dis 35:32–36
Deshmukh K, Hemrick S (1976) Metabolic changes in rabbit articular cartilage due to inflammation. Arthritis Rheum 19: 199–208
Carmichael DJ, Gillard GC, Lowther DA, Handley CJ, Santer VB (1977) Carrageenin-induced arthritis. IV. Rate changes in cartilage matrix proteoglycan synthesis. Arthritis Rheum 20:834–840
Lowther DA, Sandy JD, Santer VB, Brown HLG (1978) Antigen-induced arthritis. Decreased proteoglycan content and inhibition of proteoglycan synthesis in articular cartilage. Arthritis Rheum 21:675–680
Berg WB van den, Kruijsen MWM, Putte LBA van de, Beusekom HJ van, Sluis-van de Pol M van der, Zwarts WA (1981) Antigen-induced and zymosan-induced arthritis in mice: Studies on in vivo cartilage proteoglycan synthesis and chondrocyte death. Br J Exp Pathol 62:308–316
McKenzie LS, Horsburgh BA, Ghosh P, Taylor TKF (1976) Effect of anti-inflammatory drugs on sulphated glycosaminoglycan synthesis in aged human articular cartilage. Ann Rheum Dis 35:487–497
Palmoski MJ, Brandt KD (1979) Effect of salicylate on proteoglycan metabolism in normal canine articular cartilage in vitro. Arthritis Rheum 22:746–754
Palmoski MJ, Colyer RA, Brandt KD (1980) Marked suppression by salicylate of the augmented proteoglycan synthesis in osteoarthritic cartilage. Arthritis Rheum 23:83–91
Palmoski MJ, Brandt KD (1980) Effects of nonsteroidal anti-inflammatory drugs on proteoglycan metabolism and organization in canine articular cartilage. Arthritis Rheum 23: 1010–1020
Cole NN, Lowther DA (1969) The inhibition of chondroitin sulphate protein synthesis by cycloheximide. FEBS Lett 2:351–353
Keystone EC, Schorlemmer HU, Pope C, Allison AC (1977) Zymosan-induced arthritis. A model of chronic proliferative arthritis following activation of the alternative pathway of complement. Arthritis Rheum 20:1396–1401
Sandy J, Brown HLG, Lowther DA (1978) Degradation of proteoglycan in articular cartilage. Biochim Biophys Acta 543:536–544
Sandy J, Brown HLG, Lowther DA (1980) Control of proteoglycan synthesis. Studies on the activation of synthesis observed during culture of articular cartilage. Biochem J 188: 119–130
Herbai GA (1970) A double isotope method for determination of the miscible inorganic sulfate pool of the mouse applied to in vivo studies of sulfate incorporation into costal cartilage. Acta Physiol Scand 80:470–491
Handley CJ, Speight G, Leyden KM, Lowther DA (1980) Extracellular matrix metabolism by chondrocytes. 7. Evidence that L-glutamine is an essential amino acid for chondrocytes and other connective tissue cells. Biochim Biophys Acta 627:324–331
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van den Berg, W.B., Kruijsen, M.W.M. & van de Putte, L.B.A. The mouse patella assay. Rheumatol Int 1, 165–169 (1982). https://doi.org/10.1007/BF00541171
Issue Date:
DOI: https://doi.org/10.1007/BF00541171