Summary
Articular cartilage from 6-month-old calves was maintained in organ culture in Eagle's minimum essential medium at different oxygen tensions between 20 and 50%. When well standardized cartilage pieces of about 1 mm thickness were used the results showed a high degree of reproducibility.
The glycosaminoglycans were studied using the cetylpyridinium chloride cellulose and epichlorohydrin triethanolamine (ECTEOLA) column techniques on a microscale. In some experiments the cartilage was labelled with [35S]sulfate. Small alterations of the concentrations and distribution of different glycosaminoglycans were found and were not affected by various oxygen tensions for up to 4 weeks. A small increase was found in the fractions containing low molecular weight chondroitin sulfate. Devitalized cartilage, maintained under identical conditions, remained largely unaltered during the 4 weeks.
The [35S]sulfate activity calculated per mole of hexosamine showed a general decrease during culture maintenance. High specific activities in the fractions containing low molecular weight chondroitin sulfate and keratan sulfate were found between 1 and 3 weeks of maintenance in 20% oxygen. Maintenance in 50% oxygen resulted in severely disturbed synthesis of glycosaminoglycans.
The results are interpreted as showing that the synthesis pattern of glycosaminoglycans in articular cartilage can be altered by other factors and without the prior loss of glycosaminoglycans.
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References
Hjertquist, S-O., and R. K. Lemperg. 1972. Long term observations on the articular cartilage and autologous costal cartilage transplanted to osteochondral defects on the femoral head. Calcif. Tissue Res. 9: 226–237.
Mankin, H. J., H. Dorfman, L. Lippiello, and A. Zarins. 1971. Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J. Bone Joint Surg. [Am.] 53: 523–537.
Mankin, H. J., and L. Lippiello. 1971. The glycosaminoglycans of normal and arthritic cartilage. J. Clin. Invest. 50: 1712–1719.
Hjertquist, S-O., and R. K. Lemperg. 1972. Identification and concentration of the glycosaminoglycans of human articular cartilage in relation to age and osteoarthritis. Calcif. Tissue Res. 10: 223–237.
Bjelle, A. O., C. A. Antonopoulos, B. Engfeldt, and S-O. Hjertquist. 1972. Fractionation of glycosaminoglycans of human articular cartilage on Ecteola cellulose in ageing and in osteoarthrosis. Calcif. Tissue Res. 8: 237–246.
Bergenholtz, A. and R. K. Lemperg. 1975. Calf articular cartilage in organ culture in a chemically defined medium. I. Autoradiographic study after [35S]sulfate labeling. In Vitro 11: 286–290.
Anseth, A., C. A. Antonopoulos, A. Bjelle, and L.-Á. Fransson. 1970. Fractionation and quantitative determination of keratan sulphate using cetylpyridinium chloride and ECTEOLA-cellulose. Biochim. Biophys. Acta 215: 522–526.
Antonopoulos, C. A., S. Gardell, J. A. Szirmai, and R. De Tyssonsk. 1964. Determination of glycosaminoglycans (mucopolysaccharides) from tissues on the microgram scale. Biochim. Biophys. Acta 83: 1–19.
Antonopoulos, C. A., L-Á. Fransson, S. Gardell, and D. Heineg∢rd. 1969. Fractionation of keratan sulphate from human nucleus pulposus. Acta Chem. Scand. 23: 2616–2620.
Laurent, T. C., and J. E. Scott. 1964. Molecular weight fractionation of polyanions by cetylpyridinium chloride in salt solutions. Nature 202: 661–662.
Lemperg, R. K. 1967. Studies of autologous diced costal cartilage transplant. I. The morphological changes and35S-sulphate uptake in vitro after intramuscular implantation. Acta Soc. Med. Upsal. 72: 120–140.
Hjertquist, S-O., and R. K. Lemperg. 1970. Microchemical studies on glycosaminoglycans and calcium in autologous costal cartilage transplanted to an osteochondral defect on the femoral head of adult rabbits. Calcif. Tissue Res. 5: 153–169.
Lohmander, S., C. A. Antonopoulos, and U. Friberg. 1973. Chemical and metabolic heterogeneity of chondroitin sulphage and keratan sulphate in guinea pig cartilage and nucleus pulposus. Biochim. Biophys. Acta 304: 430–448.
Hjertquist, S-O., and L. Veilens. 1968. The glycosaminoglycans of dog compact bone and epiphyseal cartilage in the normal state and in experimental hyperparathyroidism. Calcif. Tissue Res. 2: 314–333.
Seno, N., K. Anno, K. Kondo, S. Nagase, and S. Saito. 1970. Improved method for electrophoretic separation and rapid quantitation of isomeric chondroitin sulphates on cellulose acetate strips. Anal. Biochem. 37: 197–201.
Kimura, A., and K. Tsurumi. 1969. An improved method for the electrophoretic separation of acid mucopolysaccharides on cellulose acetate sheets. J. Biochem. 65: 303–304.
Lemperg, R. K., S-E. Larsson, and S-O. Hjertquist. 1974. The glycosaminoglycans of bovine articular cartilage I. Concentration and distribution in different layers in relation to age. Calcif. Tissue Res. 15: 237–251.
Larsson, S-E., and R. K. Lemperg. 1974. The glycosaminoglycans of the different layers of bovine articular cartilage in relation to age. II. Incorporation of35S-sulphate in vitro into different fractions of chondroitin-sulphate. Calcif. Tissue Res. 15: 253–267.
Jones, I., and Lemperg, R. 1974: Chondroitin sulfate of calf knee joint cartilage. Biochem. Biophys. Acta. In Press.
Bentley, J. P., and B. Rokosová. 1970. The metabolic heterogeneity of rabbit ear cartilage chondroitin sulphate. Biochem. J. 116: 329–336.
Hardingham, T. E., and H. Muir. 1972. Biosynthesis of proteoglycans in cartilage slices. Fractionation by gel chromatography and equilibrium density-gradient centrifugation. Biochem. J. 126: 791–803.
Hjertquist, S-O., and R. K. Lemperg. 1967. Studies of autologous diced costal cartilage transplant. III. With special regard to glycosaminoglycans, hydroxyproline, calcium and35S-sulphate incorporationin vitro after intramuscular implantation. Acta Soc. Med. Upsal. 72: 173–199.
Larsson, S-E., and K. E. Kuettner. 1974. Microchemical studies of acid glycosaminoglycans from isolated chondrocytes in suspension. Calcif. Tissue Res. 14: 49–58.
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This work was supported by a grant from the Swedish Medical Research council project no. 17X-138.
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Lemperg, R.K., Bergenholtz, A. & Smith, T.W.D. Calf articular cartilage in organ culture in a chemically defined medium. In Vitro 11, 291–301 (1975). https://doi.org/10.1007/BF02615640
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DOI: https://doi.org/10.1007/BF02615640