Summary
The effect of bone matrix protein of osteonectin onde novo formation of apatite was studied in a wide range of calcium phosphate solutions in the presence of collagen. In every solution, from which amorphous calcium phosphate, octacalcium phosphate, or apatite precipitated as a possible initial phase, osteonectin at concentrations less than 1 μM retarded the precipitation, subsequent transformation to apatite, and ripening crystal growth of apatite. Collagen present as either reconstituted or denatured form had no effect on the osteonectin-associated reactions as well as osteonectin-free reactions, and no structural correlation was observed between collagen fibrils and any of the calcium phosphates that appeared in our system. Direct measurement of free calcium levels in the solutions suggested that the reduction in calcium activity due to complexing with osteonectin hardly explained the inhibitory activity of osteonectin in retarding the formation of apatite. Instead, our transmission electron microscopic (TEM) observation strongly suggested that the primary mechanism for osteonectin to inhibit the formation of apatite is to block growth sites of calcium phosphates nucleated. The apatite thus formed in the presence of osteonectin showed less resolved X-ray diffraction patterns, partly because of smaller crystallites as suggested by TEM.
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References
Robinson RA (1952) An electron-microscopic study of crystalline inorganic component of bone and its relationship to the organic matrix, J Bone Joint Surg 34a:389–434
Engstrom A, Engfeldt B, Zetterstrom R (1952) Relation between collagen and mineral salts in bone tissue. Experimentia 15:259
Francis MD, Webb NC (1971) Hydroxyapatite formation from a hydrated calcium monohydrogen phosphate precursor. Calcif Tissue Res 6:355–342
Neuman WF, Bareham BJ (1975) Evidence for the presence of secondary calcium phosphate in bone and its stabilization by acid production. Calcif Tissue Res 18:161–172
Eanes ED, Termine JD, Posner AS (1967) Amorphous calcium phosphate in skeletal tissue. Clin Orthop 53:223–235
Eanes ED, Termine JD, Nylen MU (1973) An electron microscopic study of the formation of amorphous calcium phosphate and its transformation to crystalline apatite. Calcif Tissue Res 12:143–158
Brown WE, Smith JP, Lehr JR, Frazier AW (1962) Crystallographic and chemical relations between octacalcium phosphate and hydroxyapatite. Nature 196:1048–1055
Brown WE (1966) Crystal growth of bone mineral. Clin Orthop 44:205–220
Howell DS, Pita JC, Marquez JF, Madruga JE (1968) Partition of calcium, phosphate, and protein in the fluid phase aspirated by calcifying sites in epiphyseal cartilage. J Clin Invest 47:1121–1132
Glimcher MJ, Hodge AJ, Schmitt FO (1957) Macromolecular aggregation states in relation to mineralization: the collagen-hydroxyapatite system as studied in vitro. Proc Natl Acad Sci USA 43:860–867
Solomons CC, Neuman WF (1960) On the mechanisms of calcification: the mineralization of dentin. J Biol Chem 235:2502–2506
Katz ED (1969) The kinetics of mineralization in vitro. I. The nucleator properties of 640-Å collagen at 25°C. Biochim Biophys Acta 194:121–129
Neuman WF (1980) Bone material and calcification mechanisms. In: Urist MR (ed), Fundamental and clinical bone physiology JB Lippicott, Philadelphia
Glimcher MJ (1981) On the form and function of bone from molecules to organs: Wolff's law revisited, 1981. In: Veis A (ed) The chemistry and biology of mineralized connective tissues. Elsevier North Holland, Amsterdam, New York
Urist MR, Moss MJ, Adams JM (1964) The calcification of tendon (a triphasic local mechanism). Arch Pathol 77:594–608
Wadkins CL (1968) Experimental factors that influence collagen calcification in vitro. Calcif Tissue Res 2:214–228
Bachra BN, Fisher HRA (1968) Mineral deposition in collagen in vitro. Calcif Tissue Res 2:343–352
Bachra BN (1972) Calcification in vitro of demineralized bone matrix: electron microscopic and chemical aspects. Calcif Tissue Res 8:287–303
Pocric B, Pucor Z (1979) Precipitation of calcium phosphates under conditions of double diffusion in collagen and gels of gelatin and agar. Calcif Tissue Int 27:171–176
Shuttleworth A, Veis A (1972) The isolation of anionic phosphoproteins from bovine cortical bone via the periodate solubilization of bone collagen. Biochim Biophys Acta 257:414–420
Spector AR, Glimcher MJ (1972) The extraction and characterization of soluble anionic phosphoproteins from bone. Biochim Biophys Acta 263:593–603
Hauschka PV, Lian JB, Gallop PM (1975) Direct identification of the calcium-binding amino acid, gamma-carboxyglutamate, in mineralized tissue. Proc Natl Acad Sci USA 72:3925–3927
Price PA, Otsuka AS, Poser JW, Kristaponis J, Raman N (1967) Characterization of a γ-carboxyglutamic acid-containing protein from bone. proc Natl Acad Sci USA 73:1447–1451
Termine JD, Belcourt AB, Conn KM, Kleinman HK (1981) Mineral-and collagen-binding proteins of fetal calf bone. J Biol Chem 256:10403–10408
Whitson SW, Harrison W, Dunlap MK, Bowers DE, Fisher LW, Robey PG, Termine JD (1984) Fetal bovine bone cells synthesize bone-specific matrix proteins. J Cell Biol 99:607–614
Nawrot CF, Campbell DJ, Schroeder JK, Valkenburg MW (1976) Dentin phosphoprotein-induced formation of hydroxyapatite during in vitro synthesis of amphorous calcium phosphate. Biochem 15:3445–3449
Termine JD, Kleinman HK, Whitson SW, Conn KM, McGarvey ML, Martin GR (1981) Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26:99–105
Menanteu J, Neuman WF, Neuman MW (1982) A study of bone proteins which can prevent hydroxyapatite formation. Metab Bone Dis Rel Res 4:152–162
Romberg RW, Werness PG, Lollar P, Riggs BL, Mann KG (1985) Isolation and characterization of native adult osteonectin. J Biol Chem 260:2728–2736
Romberg RW, Werness PG, Riggs BL, Mann KG (1986) Inhibition of hydroxyapatite crystal growth of bone-specific and other calcium-binding proteins. Biochem 25:1176–1180
Termine JD, Belcourt AB, Christner PJ Conn KM, Nylen MU (1980) Properties of dissociatively extracted fetal tooth matrix proteins. I. Principal molecular species in developing bovine enamel. J Biol Chem 255:9760–9768
Miller EJ, Rhodes (1982) Preparation and characterization of the different types of collagen. Methods Enzymol 82A:33–64
Termine JD, Eanes ED (1974) Calcium phosphate deposition from balanced salt solutions. Calc Tiss Res 15:81–84
Eanes ED, Meyer JL (1977) The maturation of crystalline calcium phosphates in aqueous suspensions at physiological pH. Calcif Tissue Res 23:259–269
Meyer JL, Eanes ED (1978) A thermodynamic analysis of the amorphous-to-crystalline calcium phosphate transformation. Calcif Tissue Res 25:59–68
Doi Y, Eanes ED (1984) Transmission electron microscopic study of calcium phosphate formation in supersaturated solution seeded with apatite. Calcif Tissue Int 36:39–47
Doi Y, Eanes ED, Shimokawa H, Termine JD (1984) Inhibition of seeded growth of enamel apatite crystals by amelogenin and enamelin proteins in vitro. J Dent Res 63:98–105
Shyu JL, Perez L, Zawcki SL, Heughehart JC, Nancollas GH (1981) The solubility of octacalcium phosphate at 37°C in the system Ca(OH)2-H3PO4-KNO3-H2O.
Tung MS, Brown WE (1983) An intermediate state in hydrolysis of amorphous calcium phosphate. Calcif Tissue Int 35:783–790
Chow LC, Brown WE (1984) A physicochemical benchscale caries model. J Dent Res 63:868–873
Doi Y, Eanes ED, Shimokawa H, Termine JD (1984) Modulation of seeded enamel apatite crystal growth in vitro by enamel matrix amelogenin and enamelin proteins. In: Fearnhead RW, Suga S (eds) Tooth enamel IV. Elsevier Science Publishers B.V.
Boskey AL, Posner AS (1973) Conversion of amorphous calcium phosphate to microcrystalline hydroxyapatite. A pH-dependent, solution-mediated, solid-solid conversion. J Phys Chem 77:2313–2317
Cheng P-T (1987) Formation of octacalcium phosphate and subsequent transformation to hydroxyapatite at low supersaturation: a model for cartilage calcification. Calcif Tissue Int 40:339–343
Mason IJ, Taylor A, Williams JG, Sage H, Hogan BL (1986) Evidence from molecular cloning that SPARC, a major product of mouse embryo parietal endoderm, is related to an endothelial cell ‘culture shock’ glycoprotein of Mr 43000. EMBO J 5:1465–1472
Engel J, Taylor W, Paulsson M, Sage H, Hogan B (1987) Calcium binding domains and calcium-induced conformational transition of SPARC/BM-40/osteonectin, an extracellular glycorprotein expression in mineralized and nonmineralized tissues. Biochem 26:6958–6965
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Doi, Y., Okuda, R., Takezawa, Y. et al. Osteonectin inhibitingde novo formation of apatite in the presence of collagen. Calcif Tissue Int 44, 200–208 (1989). https://doi.org/10.1007/BF02556565
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DOI: https://doi.org/10.1007/BF02556565