Abstract
The precipitation of calcium phosphatein vitro was studied in order to define the processes of homogeneous and heterogeneous nucleation and to study the effect of certain inhibitors of crystallization on these processes. It was shown that the solubility-determining surface phase of the calcium phosphate formed by homogeneous nucleation had a Ca/P molar ratio close to that of octacalcium phosphate (OCP) and that the most constant formation product in solution at the onset of spontaneous precipitation was that of OCP. Known inhibitors of crystallization had no effect on the formation product of homogeneous nucleation. Below the formation product of spontaneous precipitation nucleation was essentially heterogeneous and was markedly influenced by adding nucleating material to the system, by increasing the time of incubation, and by adding inhibitors of crystallization containing the grouping P-X-P (where X=O, C, or N). The level of supersaturation at the point of maximum inhibition was close to the formation product of spontaneous precipitation of calcium phospate. The implications of these observations are discussed in relation to the precipitation of calcium salts in urine.
Résumé
La précipitation du phosphate de calciumin vitro est étudiée dans le but de définir les processus de nucléation homogène et hétérogène et de tester l’effet de certains inhibiteurs de cristallisation dans ces processus. La phase de surface, déterminant la solubilité du phosphate de calcium, formé par nucléation homogène, a un rapport molaire Ca/P proche de celui du phosphate octocalcique (OCP) et le produit le plus constant formé, en solution au début de la précipitation spontanée, est l’OCP. Des inhibiteurs connus de cristallisation n’ont aucun effet sur le produit de formation de la nucléation homogène. Au dessous du produit de formation de précipitation spontanée, la nucléation est essentiellement hétérogène et est fortement influencée par adjonction de substances facilitant la nucléation, par augmentation du temps d’incubation et par adjonction d’inhibiteurs de cristallisation contenant le groupement P-X-P (ou X=O, C ou N). Le taux de supersaturation au point d’inhibition maximum est proche du produit de formation de la précipitation spontanée du phosphate de calcium. Les conséquences de ces observations sont envisagées en fonction de la précipitation de sels calciques dans l’urine.
Zusammenfassung
Die Fällung von Calciumphosphatin vitro wurde studiert, um die Prozesse der homogenen und der heterogenen Nukleation zu bestimmen und um die Wirkung bestimmter Kristallisationshemmer auf diese Prozesse zu untersuchen. Es wurde nachgewiesen, daß die Löslichkeits-bestimmende Oberflächenphase des Calciumphosphats, welches durch homogene Nukleation gebildet wurde, ein molares Ca/P-Verhältnis aufwies, das demjenigen von Octocalciumphosphat (OCP) sehr ähnlich war und daß das konstanteste Bildungsprodukt in der Lösung zu Beginn der spontanen Fällung dasjenige von OCP war. Bekannte Kristallisationshemmer hatten keine Wirkung auf das bildungsprodukt homogener Nukleation. Unterhalb dem Bildungsprodukt der spontanen Fällung war die Nukleation vorwiegend heterogen und wurde stark beeinflußt durch: Zufügen von nukleierendem Material zum System; Erhöhung der Inkubationszeit; und Zufügen von Kristallisationshemmern, welche die P-X-P-Gruppierung enthielten (wobei X=O, C oder N). Die Höhe der Übersättigung bei maximaler Hemmung was dem Bildungsprodukt der spontanen Fällung von Calciumphosphat sehr ähnlich. Die Bedeutung dieser Beobachtungen wird in bezug auf die Fällung von Calciumsalzen im Urin diskutiert.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bachra, B. N., Fischer, H. R. A.: The effect of some inhibitors on the nucleation and crystal growth of apatite. Calcif. Tiss. Res.3, 348–357 (1969).
Best, J. B.: Some theoretical considerations concerning crystals with relevance to the physical properties of bone. Biochim. biophys. Acta (Amst.)32, 194–202 (1959).
Boulet, M., Marier, J. R.: Precipitation of calcium phosphates from solutions at near physiological concentrations. Arch. Biochem. Biophys.93, 157–165 (1961).
Brown, W. E.: Crystal growth of bone mineral. Clin. Orthop.44, 205–220 (1966).
Eanes, E. D., Gillesen, I. H., Posner, A. S.: Intermediate states in the precipitation of hydroxyapatite. Nature (Lond.)208, 365–367 (1965).
Eanes, E. D., Posner, A. S.: Intermediate phases in the basic solution preparation of alkaline earth phosphates. Calcif. Tiss. Res.2, 38–48 (1968).
Fleisch, H., Bisaz, S.: Mechanism of calcification: inhibitory role of pyrophosphate. Nature (Lond.)195, 911 (1962).
Fleisch, H., Bisaz, S.: Isolation from urine of pyrophosphate, a calcification inhibitor. Amer. J. Physiol.203, 671–675 (1962).
Fleisch, H., Neuman, W. F.: Mechanisms of calcification: role of collagen, polyphosphates and phosphatase. Amer. J. Physiol.200, 1296–1300 (1961).
Fleisch, H., Russell, R. G. G., Bisaz, S., Mühlbauer, R. C., Williams, D. A.: The inhibitory effect of phosphonates on the formation of calcium phosphate crystalsin vitro and on aortic and kidney calcificationin vivo. Europ. J. clin. Invest.1, 12–18 (1970).
Fleisch, H., Russell, R. G., Bisaz, S., Termine, J. D., Posner, A. S.: Influence of pyrophosphate on the transformation of amorphous to crystalline calcium phosphate. Calcif. Tiss. Res.2, 49–59 (1968).
Fleisch, H., Russell, R. G. G., Straumann, F.: Effect of pyrophosphate on hydroxyapatite and its implications in calcium homeostasis. Nature (Lond.)212, 901–903 (1966).
Francis, M. D.: The inhibition of calcium hydroxyapatite crystal growth by polyphosphonates and polyphosphates. Calcif. Tiss. Res.3, 151–162 (1969).
Francis, M. D., Gray, J. A., Griebstein, W. J.: The formation and influence of surface phases on calcium phosphate solids. Advanc. oral Biol.3, 83–120 (1968).
Francis, M. D., Russell, R. G. G., Fleisch, H.: Diphosphonates inhibit formation of calcium phosphate crystalsin vitro and pathological calcificationin vivo. Science165, 1264–1266 (1969).
Francis, M. D., Webb, N. C.: Hydroxyapatite formation from a hydrated calcium monohydrogen phosphate precursor. Calcif. Tiss. Res.6, 335–342 (1971).
Füredi-Milhofer, H., Purgaric, B., Brecevic, L., Pavkovic, N.: Precipitation of calcium phosphates from electrolyte solutions. Calcif. Tiss. Res.8, 142–153 (1971).
Hirschman, A., Sobel, A. E.: Composition of the mineral deposited duringin vitro calcification in relation to the fluid phase. Arch. Biochem. Biophys.110, 237–243 (1965).
Lonsdale, K.: Epitaxy as a growth factor in urinary calculi and gallstones. Nature (Lond.)217, 56–58 (1968).
MacGregor, J., Brown, W. E.: Blood-bone equilibrium in calcium homeostasis. Nature (Lond.)205, 359–361 (1965).
Moreno, E. C., Brown, W. E., Osborn, G.: Stability of dicalcium phosphate dihydrate in aqueous solutions and solubility of octacalcium phosphate. Proc. Soil Sci. Soc. Amer.24, 99–102 (1960).
Nancollas, G. H., Mohan, M. S.: The growth of hydroxyapatite crystals. Arch. oral Biol.15, 731–745 (1970).
Neuman, W. F., Neuman, M. W.: The chemical dynamics of bone mineral. Chicago: Chicago University Press 1958.
Newesely, H.: Über die Löslichkeit schwerlöslicher Salze, insbesondere gefällter Calcium-phosphate. Mh. Chem.97, 468–483 (1966).
Nordin, B. E. C.: The estimation of “free” calcium in the urine and its relevance to calculus formation. Brit. J. Urol.31, 404–413 (1959).
Robertson, W. G.: Measurement of ionized calcium in biological fluids. Clin. chim. Acta24, 149–157 (1969).
Robertson, W. G.: A method for measuring calcium crystalluria. Clin. chim. Acta26, 105–110 (1969).
Robertson, W. G.: Physico-chemical aspects of renal stone-formation. Ph. D. Thesis, University of Leeds, 1969.
Robertson, W. G., Fleisch, H.: The effect of imidodiphosphate (P-N-P) on the precipitation and dissolution of calcium phosphatein vitro. Biochim. biophys. Acta (Amst.)222, 677–680 (1970).
Robertson, W. G., Hambleton, J., Hodgkinson, A.: Peptide inhibitors of calcium phosphate precipitation in the urine of normal and stone-forming men. Clin. chim. Acta25, 247–253 (1969).
Robertson, W. G., Nordin, B. E. C.: Renal Stone Research Symposium, eds. A. Hodgkinson and B. E. C. Nordin, p. 221. London: Churchill 1969.
Robertson, W. G., Peacock, M.: Calcium oxalate crystalluria and inhibitors of crystallisation in recurrent renal stone-formers. Clin. Sci.43, 499–506 (1972).
Robertson, W. G., Peacock, M., Nordin, B. E. G.: Activity products in stone-forming and non-stone forming urine. Clin. Sci.34, 579–594 (1968).
Robertson, W. G., Peacock, M., Nordin, B. E. C.: Calcium crystalluria in recurrent renal stone-formers. Lancet1969 II, 21–24.
Russell, R. G. G., Mühlbauer, R. C., Bisaz, S., Williams, D. A., Fleisch, H.: The influence of pyrophosphate, condensed phosphates, phosphonates and other phosphate compounds on the dissolution of hydroxyapatitein vitro and on bone resorption induced by parathyroid hormone in tissue culture and in thyroparathyroidectomised rats. Calcif. Tiss. Res.6, 183–196 (1970).
Schwartz, A.: The nucleation of calcium phosphate by organic matrices. M. Sc. Thesis, Case Institute of Technology, 1967.
Solomons, C. C., Neuman, W. F.: On the mechanisms of calcification: The remineralization of dentin. J. biol. Chem.235, 2502–2506 (1960).
Strates, B. S., Neuman, W. F., Levinskas, G. J.: The solubility of bone mineral. II. Precipitation of near-neutral solutions of calcium and phosphate. J. phys. Chem.61, 279–282 (1957).
Termine, J. D., Posner, A. S.: Calcium phosphate formationin vitro. I. Factors affecting initial phase separation. Arch. Biochem. Biophys.140, 307–317 (1970).
Walton, A. G.: The formation and properties of precipitates. New York: Interscience Publications 1967.
Walton, A. G., Bodin, W. J., Füredi, H., Schwartz, A.: Nucleation of calcium phosphate from solution. Canad. J. Chem.45, 2695–2701 (1967).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Robertson, W.G. Factors affecting the precipitation of calcium phosphatein vitro . Calc. Tis Res. 11, 311–322 (1973). https://doi.org/10.1007/BF02547230
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02547230