Summary
The metabolic, histochemical and ultrastructural modifications induced in rabbit articular cartilage during in vitro incubation at 37°C, for various periods (10 min to 18 h), using Krebs phosphate-glucose nutrient medium, were studied. It was found that after only 10 min of incubation, the chondrocytes increase their synthesis of matrix macromolecules for, at least, the next 6 h. This was suggested by:
-
1.
Increased incorporation of 35S-sulfate and 3H-glycine during the first 6 h of incubation.
-
2.
An intensification of metachromasia, which also spread out into the superficial layer that is normally orthochromatic. Only the most superficial layer corresponding to one or two rows of the cells, retained its staining pattern throughout the incubation;
-
3.
A rapidly acquired abundant rough endoplasmic reticulum, enlarged Golgi area and numerous newly synthesized proteoglycan molecules.
Similar content being viewed by others
References
Abbott, J., Holtzer, H.: The loss of phenotypic traits by differentiated cells in vitro. III — The reversible behaviour of chondrocytes in primary cultures. J. Cell Biol. 28, 473–487 (1966)
Bosmann, H.B.: Cellular control of macromolecular synthesis: rate of synthesis of extra-cellular macromolecules during and after depletion by papain. Proc. roy. Soc. B 169, 399–425 (1968)
Coelho, R.R., Chrisman, D.D.: Sulfate metabolism in cartilage. II — 35S-sulfate uptake and total sulfate in cartilage slices. J. Bone Jt. Surg. 42A, 165–172 (1960)
Collins, D.H., McElligott, T.F.: Sulfate (35SO4) uptake by chondrocytes in relation to histological changes in osteoarthritic human articular cartilage. Ann. rheum. Dis. 19, 318–330 (1960)
Curran, R.C., Gibson, T.: The uptake of labelled sulfate by human cartilage cells and its use as a test for viability. Proc. roy. Soc. B 144, 572–576 (1956)
Davies, D.V., Barnett, C.H., Cochrane, W., Palfrey, A.J.: Electron microscopy of articular cartilage in the young adult rabbit. Ann. rheum. Dis. 21, 11–21 (1962)
Fell, H.B., Barratt, M.E.J.: The role of soft connective tissue in the break-down of pig articular cartilage cultivated in the presence of complement sufficient antiserum to pig erythrocytes. I — Histochemical changes. Int. Arch. Allergy 44, 441–488 (1973)
Fitton-Jackson, S.: Environmental control of macromolecular synthesis in cartilage and bone: morphogenetic response to hyaluronidase. Proc. roy. Soc. B 175, 405–453 (1970)
Gardner, D.L.: The influence of microscopic technology on knowledge of cartilage surface structure. Ann. rheum. Dis. 31, 235–258 (1972)
Green, W.T.: Behaviour of articular chondrocytes in cell culture. Clin. Orthop. Rel. Res. 75, 248–260 (1971)
Halpin, D., Lipiello, L., Mankin, H.J.: Personal communication 1975
Lane, J.M., Brighton, C.T.: In vitro rabbit articular cartilage organ model. I — Morphology and glycosamino-glycan metabolism. Arthr. and Rheum. 17, 235–243 (1974)
Mankin, H.J.: The metabolism of articular cartilage in health and disease. In: Dynamics of connect. Tissue macromol (P.M.C. Burleigh and A.R. Poole, ed.) 327–353. London-Amsterdam-New York: North Holland and American Elsevier 1975
Maroudas, A.: Glycosamino-glycan turn-over in articular cartilage. Phil. Trans. B 271, 293–313 (1975)
McElligott, T.F., Collins, D.H.: Chondrocyte function of human articular and costal cartilage compared by measuring the in vitro uptake of labelled (35S) sulfate. Ann. rheum. Dis. 19, 31–41 (1960)
McElligott, T.F., Potter, J.L.: Increased fixation of Sulfur-35 by cartilage in vitro following depletion of the matrix by intravenous papain. J. exp. Med. 112, 743–750 (1960)
Meachim, G., Stockwell, R.A.: The matrix, the chondrocytes. In: Adult articular cartilage (Freeman, ed.) pp. 1–99, New-York: Grune and Stratton 1975
Millroy, S.J., Poole, A.R.: Pig articular cartilage in organ culture. Effect of enzymatic depletion of the matrix on response of chondrocytes to complement sufficient antiserum against pig erythrocytes. Ann. rheum. Dis. 33, 500–508 (1974)
Mitrovic, D.: Development of the metatarsophalangeal joint of the chick embryo: Morphological ultrastructural and histochemical studies. Amer. J. Anat. 150, 333–348 (1977)
Nevo, Z., Dorfman, A.: Regulation of chondromucoprotein (CMP) synthesis in chondrocytes in suspension culture. Fed. Proc. Fed. Amer. Soc. exp. Biol. 30, 3 part II (1971)
Poole, A.R.: The relationship between toluidine blue staining and hexuronic acid content of cartilage matrix. Histochem. J. 2, 425–430 (1970)
Scott, J.E., Dorling, J.: Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie 5, 221–233 (1965)
Scott, J.E., Dorling, J., Stockwell, R.A.: Reversal of protein blocking of basophilia in salt solutions: implications on the localization of polyanions using alcian blue. J. Histochem. Cytochem. 16, 383–386 (1968)
Shulman, H.J., Opler, A.: The stimulatory effect of calcium on the synthesis of cartilage proteoglycan. Biochem. biophys. Res. Commun. 59, 914–919 (1974)
Sokoloff, L., Malemud, C.J., Srivistava, V.M.L., Morgan, W.D.: In vitro culture of articular chondrocytes. Fed. Proc. 32, 1499–1502 (1973)
Szirmai, J.A., Balazs, E.A.: Metachromasia and the quantitative determination of dye-binding. Acta histochem, Suppl. 1, 56–79 (1958)
Weiss, C.: Ultrastructure caracteristics of osteoarthritis. Fed. Proc. 32, 1459–1466 (1973)
Author information
Authors and Affiliations
Additional information
This study poses fundamental questions about the mechanisms that regulate matrix synthesis by the chondrocytes.
This work was supported by INSERM and CNRS funds. Electron microscopy was done in the Laboratory for Electron Microscopy of the CNRS, 105, boulevard Raspail — 75014 Paris France. — We are grateful to Professor R. Couteaux for helpful discussions of these results and to Alan Nurden for his advice during the preparation of the manuscript
Rights and permissions
About this article
Cite this article
Mitrovic, D., Gruson, M., Demignon, J. et al. Increased metabolic activity of rabbit articular cartilage in vitro. Cell Tissue Res. 186, 149–159 (1978). https://doi.org/10.1007/BF00219661
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00219661