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Comparison of mineralization of the tibial epiphyseal plate in immature rats following treatment with cortisone, propylthiouracil or after fasting

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Abstract

Comparison of mineralization in the hypertrophic zone of the tibial epiphyseal plate in immature rats was carried out after treatment with cortisone, propylthiouracil, or after fasting. Under normal conditions, in the extracellular matrix at the calcification front, calcium and phosphate increased, sulfated mucopolysaccharides decreased, and matrix vesicles, which serve as the locus for the formation of hydroxyapatite crystals, increased. In propylthiouracil-treated rats, hydroxyapatite crystals were prominent, related to an increase in calcium deposition, a decrease of mitochondrial granules (thought to contain calcium and phosphate), an increase in the number of matrix vesicles, and to a marked decrease in the amount of sulfated mucopolysaccharide. In cortisone-treated rats, hydroxyapatite crystals were present but they were not as numerous as in the propylthiouracil-treated rats. Correspondingly, calcium deposition was slightly reduced, mitochondrial granules were more numerous than in the previous groups of rats, matrix vesicles were less numerous, and sulfated mucopolysaccharide were more prominent than in the propylthiouracil-treated rats. In fasted rats, hydroxyapatite crystals were markedly reduced or absent, and related to a decrease in calcium deposition, an increase in the number of mitochondrial granules (suggesting a delay in transport to the extracellular matrix). Matrix vesicles were markedly reduced in number, and sulfated mucopolysaccharide much more prominent than in either the cortisone or the propylthiouracil-treated rats.

Résumé

Les expériences portent sur la minéralisation de la plaque épiphysaire tibiale du rat de souche Long-Evans, étudiée après traitement à la cortisone, propylthiouracile ou après jeûn prolongé.

Dans des conditions normales, le calcium et le phosphate augmentent au niveau de la matrice extracellulaire, alors que les mucopolysaccharides sulfonés diminuent. Par contre, les vésicules de la matrice au niveau desquels se forment les cristaux d'hydroxyapatite, augmentent.

Dans les rats ayant subi un traitement à la propylthiouracile, les cristaux d'hydroxyapatite sont très apparents. Ceci est du à une augmentation du dépôt en calcium, et à une diminution des granules des mitochondries qui contiennent probablement du calcium et du phosphate. En outre, une augmentation du nombre des vésicules de la matrice est visible ainsi qu'une décroissance de la quantité des mucopolysaccharides sulfonés.

Dans les rats traités à la cortisone, les cristaux d'hydroxyapatite sont présents, mais dans une quantité moindre que dans les rats ayant subi l'effet du propylthiouracile. Le dépôt en calcium est légèrement réduit; les granules des mitochondries sont plus nombreuses que dans les groupes précédents, le nombre des vesicules de la matrice est plus faible, et les mucopolysaccharides sulfonés sont plus apparents que dans les rats traités à la propylthiouracile.

Dans les rats ayant subi l'effet du jeûn, les cristaux d'hydroxyapatite sont fortement réduits ou entièrement absents. Ceci est du à une réduction de dépôt du calcium, une augmentation du nombre des granules des mitochondries (ce qui semble indiquer que les phénomènes de transport vers la matrice extracellulaire sont ralentis), alors que les vésicules de la matrice sont présentes dans des quantités réduites. Les mucopolysaccharides sont plus apparents que dans les animaux traités à la cortisone ou à la propylthiouracile.

Zusammenfassung

Die Untersuchung beruht auf einem Vergleich der Mineralisation in der hypertrophischen Zone in der Epiphysealplatte von Long Evans Ratten die mit Kortison, Propylthiourazil oder einfachem Fasten behandelt wurden.

Unter Normalbedingungen lassen sich in der extrazellulären Matrix der Calcifikationszone die folgenden Veränderungen beobachten: der Gehalt an Calcium und Phosphat nimmt zu, derjenige an Mukopolysacchariden nimmt ab, während die Matrixvesiclen, in denen sich die Bildung des Hydroxylapatits vollzieht, zunehmen.

In Ratten die mit Propylthiourazil behandelt wurden, treten die Hydroxylapatitskristalle besonders hervor. Dies hängt mit einer Zunahme der Calciumablagerung zusamen sowie einer Abnahme der Mitochondriengranulation (in denen vermutlich Calcium und Phosphat enthalten sind). Ferner hängt damit zusammen eine numerische Zunahme der Matrixvesiceln sowie ein starke Abnahme des Gehalts an sulfonierten Mucopolysacchariden.

In den mit Kortison behandelten Ratten sind Hydroxylapatikristalle nachweisbar, wenn auch weniger zahlreich als in den mit Propylthiourazil behandelten. Dem entspricht auch eine leicht reduzierte Calciumablagerung sowie eine Mitochondrialgranulation die derjenigen der anderen Ratten überlegen ist; Matrixvesiceln sind weniger zahlreich und sulfonierte Mucopolysaccharide sind deutlicher nachweisbar als in den Tieren, die Propylthiourazil erhielten.

Fasten führt zu einem auffallenden Verlust an Hydroxylapatitkristallen. Diese können sogar nicht mehr zu erkennen sein. Dies hängt mit verminderter Calciumablagerung zusammen sowie einer Zunahme der Mitochondrialgranulation. Dies ist vermutlich Ausdruck einer Transportverzögerung zur extrazellularen Matrix. Nach Fasten ist auch die Anzahl der Matrixvesiceln auffallend herabgesetzt, und der Gehalt an sulfonierten Mucopolysacchariden ist größer als in den mit Kortison bzw. Propylthiourazil behandelten Tieren.

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References

  • Alcock, N. W.: Calcification of cartilage. Clin. Orthop.86, 287–311 (1972).

    PubMed  Google Scholar 

  • Anderson, H. C.: Vesicles in the matrix of epiphyseal cartilage: Fine structure distribution, and association with calcification.In: Proceedings of the 4th European Regional Conference on Electron Microscopy (Bocciarelli, D. S., ed.), Electron microscopy, vol. 2, p. 437–438. Rome: Tipografia Poliglotta Vaticana 1968

    Google Scholar 

  • Anderson, H. C.: Vesicles associated with calcification in the matrix of epiphyseal cartilage. J. Cell Biol.41, 59–72 (1969).

    PubMed  Google Scholar 

  • Barrett, A. J., Sledge, C. B., Dingle, J. T.: Effects of cortisol on synthesis of chondroitin sulfate by embryonic cartilage. Nature (Lond.)211, 83–84 (1966).

    Google Scholar 

  • Bernick, S., Ershoff, B. H.: Histochemical study of bone in cortisone treated rats. Endocrinology72, 231–237 (1963)

    PubMed  Google Scholar 

  • Bonucci, E.: Fine structure of early cartilage calcification. J. Ultrastruct. Res.20, 33–55 (1967)

    PubMed  Google Scholar 

  • Bonucci, E.: Further investigation on the organic/inorganic relationships in calcifying cartilage. Calc. Tiss. Res.3, 38–59 (1969)

    Google Scholar 

  • Bonucci, E.: Fine structure and histochemistry of “calcifying globules” in epiphyseal cartilage. Z. Zellforsch.103, 192–217 (1970)

    PubMed  Google Scholar 

  • Bonucci, E.: The locus of initial calcification in cartilage and bone. Clin. Orthop.86, 108–139 (1971)

    Google Scholar 

  • Campo, R. D.: Protein polysaccharide of cartilage and bone in health and disease. Clin. Orthop.68, 182–209 (1970)

    PubMed  Google Scholar 

  • Campo, R. D., Tourtellotte, C. D., Bielen, R. J.: The protein-polysaccharides of articular, epiphyseal plate and costal cartilages. Biochim. biophys. Acta (Amst.)177, 501–511 (1969).

    Google Scholar 

  • Curran, R. C., Clark, A. E., Lovell, D.: Acid mucopolysaccharides in electron microscopy. The use of the colloidal iron method. J. Anat. (Lond.)99, 427–434 (1965)

    Google Scholar 

  • Dearden, L. C.: Enhanced mineralization of the tibial epiphyseal plate in the rat following propylthiouracil treatment: A histochemical, light, and electron microscopic study. Anat. Rec. (in press)

  • Dearden, L. C., Mosier, H. D., Jr.: Electron microscopy of tibial cartilage in catch-up growth. Anat. Rec.169, 304–305 (1971)

    Google Scholar 

  • Dearden, L. C., Mosier, H. D., Jr.: Long-term recovery of chondrocytes in the tibial epiphyseal plate in rats after cortisone treatment. Clin. Orthop.87, 322–331 (1972)

    PubMed  Google Scholar 

  • Decker, J. D.: An electron microscopic investigation of osteogenesis in the embryonic chick. Amer. J. Anat.118, 591–614 (1966)

    PubMed  Google Scholar 

  • Foldes, I., Modis, L., Suveges, I.: Investigations of the mucopolysaccharides in the proximal epiphyseal cartilage of the rat: A comparison of the methods of histochemical assay. Acta morph. Acad. Sci. hung.13, 141–153 (1965).

    Google Scholar 

  • Herring, G. M.: A review of recent advances in the chemistry of calcifying cartilage and bone matrix. Calc. Tiss. Res.4 (Suppl.), 17–23 (1970)

    Google Scholar 

  • Hirschman, A., Dziewiatowski, D. D.: Protein-polysaccharide loss during endochondral ossification: Imunochemical evidence. Science154, 393–395 (1966)

    PubMed  Google Scholar 

  • Kunin, A. S., Meyer, W. L.: The effect of cortisone on intermediary metabolism of epiphyseal cartilage from rats. Arch. Biochem. Biophys.129, 421–430 (1969)

    PubMed  Google Scholar 

  • Lillie, R. D.: Histopathological technic and practical histochemistry, 3rd ed. New York: McGraw-Hill (1965)

    Google Scholar 

  • Marinozzi, V.: Silver impregnation of ultra thin sections for electron microscopy. J. biophys. biochem. Cytol.9, 121–132 (1961)

    PubMed  Google Scholar 

  • Martin, J. H., Matthews, J. L.: Mitochondrial granules in chrondrocytes. Calc. Tiss. Res.3, 184–193 (1969)

    Google Scholar 

  • Matthews, J. L., Martin, J. H., Sampson, H. W., Kunin, A. S., Roan, J. H.: Mitochondrial granules in the normal and rachitic rat epiphysis. Calc. Tiss. Res.5, 91–99 (1970)

    Google Scholar 

  • Matukas, V. J., Panner, B. J., Orbison, J. L.: Studies on ultrastructural identification and distribution of protein-polysaccharide in cartilage matrix. J. Cell Biol.32, 365–377 (1967)

    PubMed  Google Scholar 

  • Mosier, H. D., Jr.: Allometry of body weight and tail length in studies of catch-up growth in rats. Growth33, 319–330 (1969)

    PubMed  Google Scholar 

  • Mosier, H. D., Jr.: Failure of compensatory (catch-up) growth in the rat. Pediat. Res.5, 59–63 (1971)

    Google Scholar 

  • Mowry, R. W.: Improved procedure for the staining of acidic polysacharides by Müllers colloidal (hydrous) ferric oxide and its combination with the Feulgen and periodic acid-Schiff reactions. Lab. Invest.7, 566–576 (1958)

    PubMed  Google Scholar 

  • Rambourg, A., Hernandez, W., Leblond, C. P.: Detection of complex carbohydrates in the Golgi apparatus of rat cells. J. Cell. Biol.40, 395–414 (1969)

    PubMed  Google Scholar 

  • Reynolds, E. S.: The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol.17, 208–212 (1963)

    PubMed  Google Scholar 

  • Rosenquist, T. H., Slavin, B. G., Bernick, S.: The Pearson silver-gelatin method for light microscopy of 0.5–2 μ plastic sections. Stain Technol.5, 253–257 (1971)

    Google Scholar 

  • Schrywer, H. F.: The effect of hydrocortisone on chondroitin sulfate production and loss by embryonic chick tibiotarsi in organ culture. Exp. Cell Res.40, 610–618 (1965)

    PubMed  Google Scholar 

  • Schubert, M., Pras, M.: Ground substance protein polysaccharides and precipitation of calcium phosphate. Clin. Orthop.60, 235–255 (1968)

    PubMed  Google Scholar 

  • Shapiro, I. M., Greenspan, J. S.: Are mitochondria directly involved in biological mineralization? Calc. Tiss. Res.3, 100–102 (1969)

    Google Scholar 

  • Smith, J. W.: The disposition of protein polysaccharide in the epiphyseal plate cartilage of the young rabbit. J. Cell Sci.6, 843–864 (1970)

    PubMed  Google Scholar 

  • Spicer, S. S.: Histochemical differentiation of sulfated rodent mucins. Ann. Histochem.7, 23–28 (1962)

    Google Scholar 

  • Watson, M. L.: Staining of tissue sections for electron microscopy with heavy metals. J. biophys. biochem. Cytol.4, 475 (1958)

    PubMed  Google Scholar 

  • Wuthier, R. E.: A zonal analysis of inorganic and organic constituents of the epiphysis during endochondral calcification. Calc. Tiss. Res.4, 20–38 (1969).

    Google Scholar 

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Dearden, L.C., Espinosa, T. Comparison of mineralization of the tibial epiphyseal plate in immature rats following treatment with cortisone, propylthiouracil or after fasting. Calc. Tis Res. 15, 93–110 (1974). https://doi.org/10.1007/BF02059048

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