Abstract
Mineral from medullary bone of three avian species was examined with the electron microscope in order to clarify the ultrastructure of amorphous bone mineral (ABM) in a mineralized tissue. Powders from freeze-dried bone revealed bone mineral with morphology and behavior identical to synthetic amorphous calcium phosphate (ACP). These powders exhibited spherically shaped particles 180–400 Å in diameter with uniform electron density when viewed at low bem intensity. Thin sections of embedded freeze-dried bone also revealed spherically shaped particles 100–350 Å in diameter with electron beam sensitivity characteristic of ACP. Regions of bone mineral with irregular outline and morphology were observed which closely resemble the discoidal form of synthetic ACP. More electron-dense spherical particles (150 Å in diameter) could be seen budding from these regions. Some of these buds exhibited electronlucent centers characteristics of AMB. The inorganic nature of these features of bone mineral was confirmed using ultramicroincineration. Preliminary exploration of a freeze-substitution technique showed spherical bone mineral particles which were similar in morphology to those observed in freeze-dried samples. A limited degree of preservation of cellular material was observed using this freeze-substitution technique.
Similar content being viewed by others
References
Anderson, C. E., Parker, J.: Electron microscopy of the epiphyseal cartilage plate. Clin. Orthop.58, 225–241 (1968)
Arnott, H. J., Pautard, F. G. E.: Osteoblast function and fine structure. Israel J. med. Sci.3, 657–670 (1967)
Bachra, B. N., Trautz, O. R., Simon, S. L.: Precipitation of calcium carbonates and phosphates. III. The effect of magnesium and fluoride ions on the spontaneous precipitation of calcium carbonates and phosphates. Arch. oral. Biol.10, 731–738 (1965)
Boothroyd, B.: The problem of demineralization in thin sections of fully calcified bone. J. Cell Biol.20, 165–173 (1964)
Eanes, E. D., Termine, J. D., Nylen, M. U.: An electron microscopic study of the formation of amorphous calcium phosphate and its transformation to crystalline apatite. Calcif. Tiss. Res.12, 143–158 (1973)
Eanes, E. D., Termine, J. D., Posner, A. S.: Amorphous calcium phosphate in skeletal tissues. Clin. Orthop.53, 223–235 (1967)
Frazier, P. D.: An electron microscopic investigation of mineralizing tissues. Ph. D. dissertation, University of Washington 1971
Frazier, P. D., Brown, F. J., Rose, L. S., Fowler, B. O.: Radiofrequency oxygen excitation apparatus for low temperature ashing. J. dent. Res.46, 1098–1101 (1967)
Gersch, I.: Relation of the walls of large matrix compartments of epiphyseal cartilage to the formation of calcium crystals. In: Submicroscopic cytochemistry. II. Membranes, mitochondria and connective tissue (Gersch, I., ed.), p. 187–205. New York: Academic Press 1973
Harper, R. A., Posner, A. S.: Measurement of noncrystalline calcium phosphate in bone material. Proc. Soc. exp. Biol. (N.Y.)122, 137–142 (1966)
Hawkes, J. A. W.: Aspects of the molt pattern, calcium fluctuation, and ultrastructure of the integument in the fresh-water Isopod,Lirceus brachyurus, Harger. Ph. D. dissertation, The Pennsylvania State University 1971
Hohman, W., Schraer, H.: Low temperature ultramicroincinceration of thin-sectioned tissue. J. Cell Biol.55, 328–354 (1972)
Molnar, Z.: Development of the parietal bone of young mice. I. Crystals of bone mineral in frozen-dried preparations. J. Ultrastruct. Res.8, 491–505 (1959)
Nylen, M. U., Eanes, E. D., Termine, J. D.: Molecular and ultrastructural studies of non-crystalline calcium phosphates. Calcif. Tiss. Res.9, 95–108 (1972)
Pease, O. C.: Eutectic ethylene glycol and pure propylene glycol as substituting media for the dehydration of frozen tissue. J. Ultrastruct. Res.21, 75–97 (1967)
Quinaux, N., Richelle, L. J.: X-ray diffraction and infrared analysis of bone; specific gravity fractions in the growing rat. Israel J. med. Sci.3, 667–690 (1967)
Robinson, R. A., Watson, M. L.: Crystal-collagen relationship in bone as observed in the electron microscope. III. Crystal and collagen morphology as a function of age. Ann. N.Y. Acad. Sci.60, 596–628 (1955)
Schraer, H., Tannenbaum, P. J., Posner, A. S.: Crystalline changes in avian bone related to the reproductive cycle. J. dent. Res.46, 1072–1074 (1967)
Sheldon, H., Robinson, R. A.: Electron microscope studies of crystal-collagen relationships in bone. IV. The occurrence of crystals within collagen fibrils. J. biophys. biochem. Cytol.3, 1011–1015 (1957)
Spurr, A. R.: A low viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res.26, 31–43 (1969)
Tannenbaum, P. J., Schraer, H., Posner, A. S.: Crystalline changes in avian bone related to the reproductive cycle. II. Percent crystallinity changes. Calcif. Tiss. Res.14, 83–86 (1974)
Termine, J. D., Posner, A. S.: Infrared analysis of rat bone: Age dependency of amorphous and crystalline mineral fractions. Science153, 1523–1525 (1966)
Termine, J. D., Wuthier, R. E., Posner, A. S.: Amorphous-crystalline mineral changes during endochondral and periosteal bone formation. Proc. Soc. exp. Biol. (N.Y.)125, 4–9 (1967)
Weber, J. C., Eanes, J. D., Gerdes, R. J.: Electron microscope study of non-crystalline calcium phosphate. Arch. Biochem. Biophys.120, 723–724 (1967)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miller, A.L., Schraer, H. Ultrastructural observations of amorphous bone mineral in avian bone. Calc. Tis Res. 18, 311–324 (1975). https://doi.org/10.1007/BF02546249
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02546249