Abstract
The organic-inorganic relationships in bone matrix undergoing osteoclastic resorption have been studied in rat tibial diaphyses using electron microscope techniques in an attempt to identify the steps of the resorption process. Results suggest that bone resorption occurs in two phases: the first, an extracellular phase, leads to bone matrix fragmentation and partial dissolution, and the second, an intracellular phase, to complete digestion of the breakdown products of the bone matrix. The first component of the bone matrix to be attacked by the osteoclast is the ground substance. This induces the release of the crystals lying between, and on, the collagen fibrils; any crystals lying within fibrils are released later, when the fibrils break up. As this stage proceeds, the collagen fibrils retain their normal intrinsic texture, but gradually loose their lateral aggregation, appearing as individual fibrils (some of them uncovered by crystals), mixed with fragments of fibrils and many free crystals. The loosened but otherwise structurally normal collagen fibrils, and their fragments, are strongly argyrophilic. Complete dissolution of the disaggregated fibrils occurs outside the cell, both in the resorption zone and in the initial portion of the channels of the ruffled border. The free crystals present in the resorption zone and those phagocytosed in cytoplasmic vacuoles are organic-inorganic structures, whose organic component (the crystal ghost) is, at least in part, of proteoglycan nature. Dissolution of inorganic material occurs within the cytoplasmic vacuoles of the osteoclasts. Results are viewed in relation to the process of bone resorption and, as far as crystal ghosts are concerned, to that of bone calcification. A tentative summary of the various steps involved in the mechanism of bone resorption is given.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Appleton, J.: Ultrastructural observations on early cartilage calcification. The use of chromium sulphate in decalcification. Calc. Tiss. Res.5, 270–276 (1970)
Appleton, J.: Ultrastructural observations on the inorganic/organic relationships in early cartilage calcification. Calc. Tiss. Res.7, 307–317 (1971)
Appleton, J., Balckwood, H. J. J.: Ultrastructural observations on early mineralization in cartilage. J. Bone Jt. Surg.51B, 385 (1969)
Ascenzi, A.: The relationship between mineralization and bone matrix. In: Bone and tooth (ed. H. J. J. Blackwood) p. 231–243. Oxford: Pergamon Press 1964
Ascenzi, A., Bonucci, E., Steve Bocciarelli, D.: An electron microscope study of osteon calcification. J. Ultrastruct. Res.12, 287–303 (1965)
Babai, F., Bernhard, W.: Détection cytochimique par l'acide phosphotungstique de certains polysaccharides sur coupes à congélation ultrafines. J. Ultrastruct. Res.37, 601–617 (1971)
Bernard, G. W., Pease, D. C.: An electron microscopic study of initial intramembranous osteogenesis. Amer. J. Anat.125, 271–290 (1969)
Bonucci, E.: Fine structure of early cartilage calcification. J. Ultrastruct. Res.20, 33–50 (1967)
Bonucci, E.: Further investigation on the organic/inorganic relationships in calcifying cartilage. Calc. Tiss. Res.3, 38–54 (1969)
Bonucci, E.: Fine structure and histochemistry of “calcifying globules” in epiphyseal cartilage. Z. Zellforsch.103, 192–217 (1970)
Bonucci, E.: The locus of initial calcification in cartilage and bone. Clin. Orthop.78, 108–139 (1971a)
Bonucci, E.: Problemi attuali attinenti all'istochimica di talune matrici calcificanti normali e patologiche. Riv. Istochim. Norm. Patol.17, 153–234 (1971b)
Bonucci, E.: Organic-inorganic relationships in calcified organic matrices. In: Colloque international sur la physicochimie et la cristallographie des apatites d'interet biologique (ed. G. Montel). (in press)
Bonucci, E., Derenzini, M., Marinozzi, V.: The organic-inorganic relationship in calcified mitochondria. J. Cell Biol.59, 185–211 (1973)
Cameron, D. A.: The fine structure of bone and calcified cartilage. Clin. Orthop.26, 199–228 (1963)
Cameron, D. A.: The ultrastructural basis of resorption. Calc. Tiss. Res.4, 279–280 (1969)
Cameron, D. A.: The ultrastructure of bone. In: The biochemistry and Physiology of bone, 2nd Ed., vol. I (ed. G. H. Bourne), p. 191–236. New York and London: Academic Press 1972
Cohn, Z. A., Fedorko, M. E.: The formation and fate of lysosomes. In: Lysosomes in biology and pathology, vol. I (ed. J. T. Dingle, H. B. Fell), p. 43–63. Amsterdam: North Holland 1969
Decker, J. D.: An electron microscopic investigation of osteogenesis in the embryonic chick. Amer. J. Anat.118, 591–614 (1966)
Dingle, J. T.: Lysosomal enzymes in skeletal tissues. In: Hard tissue growth, repair and remineralization, Ciba Found. Symp. 11, p. 295–311. Amsterdam: Elsevier-Excerpta Medica-North-Holland 1973
Dixon, J. S., Hunter, J. A. A., Steven, F. S.: An electron microscopic study of the effect of crude bacterial α-amylase and ethylenediaminetetraacetic acid on human tendon. J. Ultrastruct. Res.38, 466–472 (1972)
Doty, S. B., Schofield, B. H.: Electron microscopic localization of hydrolytic enzymes in osteoclasts. Histochem. J.4, 245–258 (1972)
Dudley, H. R., Spiro, D.: The fine structure of bone cells. J. biophys. biochem. Cytol.11, 627–649 (1961)
Fainstat, T.: Extracellular studies of uterus I. Disappearance of the discrete collagen bundles in endometrial stroma during various reproduction states in the rat. Amer. J. Anat.112, 337–369 (1963)
Freilich, L. S.: Ultrastructure and acid phosphatase cytochemistry of odontoclasts: effects of parathyroid extract. J. Dent. Res.50, 1047–1055 (1971)
Glimcher, M. J., Krane, S. M.: The organization and structure of bone, and the mechanism of calcification. In: Treatise on collagen, vol. II B, Biology of collagen (ed. B. S. Gould), p. 67–251. London and New York: Academic Press 1968
Gonzales, F., Karnovsky, M. J.: Electron microscopy of osteoclasts in healing fractures of rat bone. J. biophys. biochem. Cytol.9, 299–316 (1961)
Göthlin, G., Ericsson, J. L. E.: Observations on the mode of uptake of thorium dioxide particles by osteoclasts in fracture callus. Calc. Tiss. Res.10, 216–222 (1972)
Hancox, N. M.: Biology of bone. Cambridge: Cambridge University Press 1972
Hancox, N. M., Boothroyd, B.: Motion picture and electron microscope studies on the embryonic avian osteoclast. J. biophys. biochem. Cytol.11, 651–661 (1961)
Hancox, N. M., Boothroyd, B.: Structure-function relationship in the osteoclast. In: Mechanisms of hard tissue destruction (ed. R. F. Sognnaes), p. 497–514. Washington: American Association for the Advancement of Science 1963
Höhling, H. J., Kreilos, R., Neubauer, G., Boyde, A.: Electron microscopy and electron microscopical measurements of collagen mineralization in hard tissues. Z. Zellforsch.122, 36–52 (1971)
Iglesias, J. R., Bernier, R., Simard, R.: Ultracryotomy a routine procedure. J. Ultrastruct. Res.36, 271–289 (1971)
Jackson, D. S., Bentley, J. P.: Collagen-glycosaminoglycans interactions. In: Treatise on Collagen, vol. II A: Biology of collagen (ed. B. S. Gould), p. 189–214. London and New York: Academic Press 1968
Jenkins, G. N., Dawes, C.: The possible role of chelation in decalcification of biological systems. In: Mechanism of hard tissue destruction (ed. R. F. Sognnaes), p. 637–662. Washington: American Association for the Advancement of Science 1963
Kallio, D. M., Garant, P. R., Minkin, C.: Evidence of coated membranes in the ruffled border of the osteoclast. J. Ultrastruct. Res.37, 169–177 (1971)
Knese, K.-H.: Osteoklasten, Chondroklasten, Mineraloklasten, Kollagenoklasten. Acta anat. (Basel)83, 275–288 (1972)
Kobayashi, T. K., Pedrini, V.: Proteoglycans-collagen interactions in human costal cartilage. Biochim. biophys. Acta (Amst.)303, 148–160 (1973)
Lowther, D. A., Natarajan, M.: The influence of glycoprotein on collagen fibril formation in the presence of chondroitin sulphate proteoglycan. Biochem. J.127, 607–608 (1972)
Lucht, U.: Acid phosphatase of osteoclasts demonstrated by electron microscopic histochemistry. Histochemie28, 103–117 (1971)
Lucht, U.: Absorption of peroxidase by osteoclasts as studied by electron microscope histochemistry. Histochemie29, 274–286 (1972a)
Lucht, U.: Osteoclasts and their relationship to bone as studied by electron microscopy. Z. Zellforsch.135, 211–228 (1972b)
Lucht, U.: Cytoplasmic vacuoles and bodies of the osteoclast. An electron microscope study. Z. Zellforsch.135, 229–244 (1972c)
Malkani, K., Luxembourger, M.-M., Rebel, A.: Cytoplasmic modifications at the contact zone of osteoclasts and calcified tissue, in the diaphyseal growing plate of foetal guinea-pig tibia. Calc. Tiss. Res.11, 258–264 (1973)
Marinozzi, V.: Cytochimie ultrastructurale du nucléole-RNA et protéines intranucléolaires. J. Ultrastruct. Res.10, 433–456 (1964)
Marinozzi, V.: Réaction de l'acide phosphotungstique avec la mucin et les glycoprotéines des plasmamembranes. J. Microscopie6, 68a (1967)
Marinozzi, V.: Phosphotungstic acid (PTA) as a stain for polysaccharides and glycoproteins in electron microscopy. In: Electron microscopy 1968, vol. II (ed. D. S. Bocciarelli), p. 55–56. Rome: Tipografia Poliglotta Vaticana 1968
Mathews, M. B.: The interactions of proteoglycans and collagen. Model systems. In: Chemistry and molecular biology of the intercellular matrix, vol. II (ed. E. A. Balazs), p. 1155–1169. London and New York: Academic Press 1970
Nisbet, J. A., Helliwell, S., Nordin, B. E. C.: Relation of lactic and citric acid metabolism to bone resorption in tissue culture. Clin. Orthop.70, 220–230 (1970)
Nylen, M. U., Scott, D. B., Mosley, V. M.: Mineralization of turkey leg tendon. II. Collagen-mineral relations revealed by electron and X-ray microscopy. In: Calcification in biological systems (ed. R. F. Sognnaes), p. 129–142. Washington: American Association for the Advancement of Science 1960
Pease, D. C.: Polysaccharides associated with the exterior surface of epithelial cells: kidney, intestine, brain. J. Ultrastruct. Res.15, 555–588 (1966)
Pease, D. C.: Phosphotungstic, acid as a specific electron stain for complex carbohydrates. J. Histochem. Cytochem.18, 455–458 (1970)
Pease, D. C., Bouteille, M.: The tridimensional ultrastructure of native collagenous fibrils cytochemical evidence for a carbohydrate matrix. J. Ultrastruct. Res.35, 339–358 (1971)
Quintarelli, G., Dellovo, M. C., Balduini, C., Castellani, A. A.: The effects of alpha amylase on collagen-proteoglycans and collagen-glycoprotein complexes in connective tissue matrices. Histochemie18, 373–375 (1969)
Rambourg, A.: Détection des glycoprotéines en microscopie, électronique par l'acide phosphotungstique à bas pH. In: Electron microscopy 1968, vol. II (ed. D. S. Bocciarelli), p. 57–58. Rome: Tipografia Poliglotta Vaticana 1968
Rambourg, A.: Localisation ultrastructurale et nature du matériel coloré au niveau de la surface cellulaire par le mélange chromique-phosphotungstique. J. Microscopie8, 325–342 (1969)
Rambourg, A.: Morphological and histochemical aspects of glycoproteins at the surface of animal cells. Int. Rev. Cytol.31, 57–114 (1971)
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)
Robinson, R. A., Cameron, D. A.: Electron microscopy of cartilage and bone matrix at the distal epiphyseal line of the femur in the newborn infant. J. biophys. biochem. Cytol.2 (Suppl.), 253–260 (1956)
Schenk, R. K., Spiro, D., Wiener, J.: Cartilage, resorption in the tibial epiphyseal plate of growing rats. J. Cell Biol.34, 275–291 (1967)
Scherft, J. P.: The resorption of the organic matrix of calcified cartilage as seen with the electron microscope. Calc. Tiss. Res.2 (suppl.), 96–96B (1968)
Scott, B. L.: The occurrence of specific cytoplasmic granules in the osteoclast. J. Ultrastruct. Res.19, 417–431 (1967)
Scott, B. L., Pease, D. C.: Electron microscopy of the epiphyseal apparatus. Anat. Rec.126, 465–495 (1956)
Smith, J. W.: The disposition of proteinpolysaccharide in the epiphyseal plate cartilage of the young rabbit. J. Cell Sci.6, 843–864 (1970)
Steven, F. S.: The Nishihara technique for the solubilization of collagen. Application to the preparation of soluble collagens from normal and rheumatoid connective tissue. Ann. Rheum. Dis.23, 300–301 (1964)
Sundström, B., Takuma, S.: A further contribution on the ultrastructure of calcifying cartilage. J. Ultrastruct. Res.36, 419–424 (1971)
Takuma, S.: Electron microscopy of the developing cartilaginous epiphysis. Arch. Oral Biol.2, 111–119 (1960)
Vaes, G.: On the mechanism of bone, resorption. J. Cell Biol.39, 676–697 (1968)
Vaughan, J. M.: The physiology of bone. Oxford: Clarendon Press 1970
Walker, D. G.: Enzymatic and electron microscopic analysis of isolated osteoclasts. Calc. Tiss. Res.9, 296–309 (1972)
Weinstock, A.: Elaboration of enamel and dentin matrix glycoproteins. In: The biochemistry and physiology of bone, 2nd ed., vol. II (ed. G. H. Bourne), p. 121–154. New York and London: Academic Press 1972
Weinstock, A., Leblond, C. P.: Elaboration of the matrix glycoprotein of enamel by the secretory ameloblasts of the rat incisor as revealed by radioautography after galactose-3H injection. J. Cell Biol.51, 26–51 (1971)
Woessner, J. F., Jr.: Biological mechanisms of collagen resorption. In: Treatise on collagen, vol. II (ed. B. S. Gould), p. 253–330. London and New York: Academic Press 1968
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bonucci, E. The organic-inorganic relationships in bone matrix undergoing osteoclastic resorption. Calc. Tis Res. 16, 13–36 (1974). https://doi.org/10.1007/BF02008210
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02008210