Abstract
Two diphosphonates, disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) and disodium dichloromethyle diphosphonate (Cl2MDP), inhibit cell-mediated bone resorption of mouse calvaria cultivated for 48 hoursin vitro, when the compounds are added to the medium. Cl2MDP is more effective than EHDP over the dose range 0–16 μg P/ml. Pyrophosphate and imidodiophosphate do not block bone resorption at comparable dose levels. When the two diphosphonates are injected into micein vivo before explants are prepared, subsequent bone resorptionin vitro is considerably reduced; at a dose level of 10 μg P/g body weight of Cl2MDP it is almost completely blocked. This effect is rapid and persists for several days. The implications of these results and the method of testing inhibitors of bone resorption by the combinedin vivo/in vitro method are discussed.
Résumé
Deux diphosphonates, le disodium-éthane-1-hydroxyle-1,1-diphosphonate (EHDP) et le disodium dichlorométhylène diphosphonate (Cl2MDP), inhibent la résorption osseuse, induite par des cellules au niveau de calottes craniennes, cultivées pendant 48 heuresin vitro, lorsque ces substances sont ajoutées au milieu. Le Cl2MDP est plus actif que l'EHDP, à des doses variant 0–16 μg P/ml. Le pyrophosphate et l'imidodiphosphate n'inhibent pas la résorption osseuse à des doses comparables. Lorsque les deux diphosphonates sont injectés à des sourisin vivo avant mise en culture, la résorption osseuse observéein vitro est considérablement réduite: à une dose de 10 μg P/g de poids corporel de Cl2MDP, elle est presque totalement inhibée. Cet effet est rapide et dure plusieurs jours. Les conséquences de ces résultats et la méthode d'essai d'inhibiteurs de la résorption osseuse par la méthode combinéein vivo/ in vitro sont envisagées.
Zusammenfassung
Zwei Diphosphonate, Dinatrium-äthan-1-hydroxy-1,1-diphosphonat (EHDP) und Dinatrium-Dichloromethylendiphosphonat (Cl2MDP), hemmen zellbedingte Knochenresorption von Mäuseschädeldächern, welche während 48 Stdin vitro kultiviert worden waren, wenn diese Substanzen dem Nährmedium zugegeben werden. Im Dosierungsbereich von 0–16 μg P/ml ist Cl2MDP wirksamer als EHDP. Pyrophosphat und Imidodiphosphat blockieren die Knochenresorption bei entsprechenden Dosen nicht. Wenn die zwei Diphosphonate Mäusenin vivo injiziert werden, bevor das Explantat hergestellt wird, ist die nachfolgende Knochenresorptionin vitro stark vermindert; bei einer Dosierung von 10 μg P/g Körpergewicht von Cl2MDP ist die Resorption fast gänzlich blockiert. Diese Wirkung erfolgt rasch und dauert während einigen Tagen an. Die Folgerungen aus diesen Ergebnissen sowie das Verfahren, Knochenresorptionshemmer mittels kombinierterin vivo/in vitro-Methode zu prüfen, werden diskutiert.
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References
Au, W. Y. W., Bartter, F. C.: Effect of vitamin D onin vitro bone calcium metabolism. Endocrinology78, 1100–1104 (1966).
Bijvoet, O. L. M., Sluys Veer, J. van der, Wildiers, J., Smeenk, D.: Effects of long term calcitonin administration to patients. In: Calcitonin 69, p. 531–539 (Taylor, S., Foster, G. V., eds.). London: William Heinemann Medical Books 1970.
Borle, A. B., Nichols, N., Nichols, G.: Metabolic studies of bonein vitro. J. biol. Chem.235, 1211–1214 (1960).
Cooper, C. W., Yates, C. W., Talmage, R. V.: Some endogenous PTH effects manifested by bonein vitro. Proc. Soc. exp. Biol. (N.Y.)119, 81–88 (1965).
Flanagan, B., Nichols, G.: Parathyroid inhibition of bone collagen synthesis. Endocrinology74, 180–186 (1964).
Fleisch, H., Bonjour, J.-P., Morgan, D. B., Reynolds, J. J., Schenk, R., Smith, R., Russell, R. G. G.: Diphosphonates. In: Endocrinology 1971. London: William Heinemann Medical Books (in press).
Fleisch, H., Maerki, J., Russell, R. G. G.: Effect of pyrophosphate on dissolution of hydroxyapatite and its possible importance in calcium homeostasis. Proc. Soc. exp. Biol. (N.Y.)122, 317–320 (1966a).
Fleisch, H., Russell, R. G. G., Straumann, F.: Effect of pyrophosphate on hydroxyapatite and its implications in calcium homeostasis. Nature (Lond.)212, 901–903 (1966b).
Fleisch, H., Russell, R. G. G., Bisaz, S. Casey, P. A., Mühlbauer, R. C.: The influence of pyrophosphate analogues (diphosphonates) on the precipitation and dissolution of calcium phosphatein vivo andin vitro. Calc. Tiss. Res.2, Suppl. 10-10A (1968).
Fleisch, H., Russell, R. G. G., Francis, M. D.: Diphosphonates inhibit hydroxyapatite dissolutionin vitro and bone resorption in tissue culture andin vivo. Science165, 1262–1264 (1969a).
Fleisch, H., Russell, R. G. G., Simpson, B., Mühlbauer, R. C.: Prevention by a diphosphonate of immobilisation osteoporosis in rats. Nature (Lond.)223, 211–212 (1969b).
Francis, M. D.: The inhibition of calcium hydroxyapatite growth by polyphosphonates and polyphosphates. Calcif. Tiss. Res.3, 161–162 (1969).
Gasser, A. B., Morgan, D. B., Fleisch, H. A., Richelle, L. J.: The influence of two diphosphonates on calcium metabolism in the rat. Clin. Sci.43, 31–45 (1972).
Johnston, C. C., Deiss, W. P., French, R. S.: Effects of changes in parathyroid status and calcium equilibrium on bone matrix metabolism. Proc. Soc. exp. Biol. (N.Y.)118, 551–554 (1965).
Jung, A., Bisaz, S., Fleisch, H.: The binding of pyrophosphate and diphosphonates to hydroxyapatite crystals. Submitted.
Kennedy, B. J.: Metabolic and toxic effects of mithramycin during tumour therapy. Amer. J. Med.49, 494–503 (1970).
Michael, W. R., King, W. R., Francis, M. D.: Effectiveness of diphosphonates in preventing osteoporosis of disuse in the rat. Clin. Orthop. (in press).
Mühlbauer, R. C., Russell, R. G. G., Williams, D. A., Fleisch, H.: The effects of diphosphonates, polyphosphates, and calcitonin on “immobilisation osteoporosis” in rats. Europ. J. clin. Invest.1, 336–344 (1971).
Raisz, L. G., Niemann, I.: Effect of phosphate, calcium and magnesium on bone resorption and hormonal responses in tissue culture. Endocrinology85, 446–452 (1969).
Reynolds, J. J.: Use of bone cultures in studying phosphate metabolism. In: Phosphate et metabolisme phosphocalcique, p. 47–55 (D. J. Hioco, ed.). Paris: Sandoz Editions 1971.
Reynolds, J. J.: A sensitivein vivo/in vitro method for studying substances that influence the resorption of bone. In: Parathyroid hormone and the calcitonins; Proceedings of an International Symposium at Chapel Hill., p. 454–462, Amsterdam: Excerpta Medica Foundation 1972.
Reynolds, J. J., Dingle, J. T.: A sensitivein vitro method for studying the induction and inhibition of bone resorption. Calc. Tiss. Res.4, 339–349 (1970).
Reynolds, J. J., Minkin, C.: Bone studiesin vitro: use of calcitonin as a specific inhibitor of bone resorption. In: Calcitonin 69, p. 168–174 (Taylor, S., Foster, G. V., eds.). London: William Heinemann Medical Books 1970.
Reynolds, J. J., Morgan, D. B.: A combinedin vivo/in vitro study of the effects of diphosphonates on bone resorption. J. Bone Jt Surg. B52, 796–797 (1970).
Robertson, W. G., Fleisch, H.: The effect of imidodiphosphonate (P−N−P) on the precipitation and dissolution of calcium phosphatein vitro. Biochim. biophys. Acta (Amst.)22, 677–680 (1970).
Robertson, W. G., Morgan, D. B., Fleisch, H., Francis, M. D.: The effects of diphosphonates on the exchangeable and non-exchangeable calcium and phosphate of hydroxyapatite. (In preparation).
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 in tissue culture and in thyroparathyroidectomised rats. Calc. Tiss. Res.6, 183–196 (1970).
Smith, R., Russell, R. G. G., Bishop, M.: Diphosphonates and Paget's disease of bone. LancetI, 945–947 (1971).
Woodhouse, N. J. Y., Bordier, P., Fisher, M., Joplin, G. F., Reiner, M., Kalu, D. N., Foster, G. V., MacIntyre, I.: Human calcitonin in the treatment of Paget's bone disease. LancetI, 1139–1143 (1971).
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Reynolds, J.J., Minkin, C., Morgan, D.B. et al. The effect of two diphosphonates on the resorption of mouse calvariain vitro . Calc. Tis Res. 10, 302–313 (1972). https://doi.org/10.1007/BF02012561
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DOI: https://doi.org/10.1007/BF02012561