Summary
Fetal rat bones were cultured in either growth-inducing or resorption-inducing media to study mineral losses during bone growth and atrophy in vitro. Whole radii and ulnae from 19-day-old fetal rats, prelabeled with45Ca and/or3H-tetracycline, were cultured intact or cut, and then digested by collagenase to obtain the calcified portion of the bones. Three-to five-fold more3H-tetracycline than45Ca was lost from the calcified portion when the bones were cultured for 4 days in growth-inducing media. Similar small amounts of45Ca were lost from live and killed bones, but more3H-tetracycline was lost from live bones than from killed bones. More3H-tetracycline was released into the growth medium with a low concentration of calcium (0.5 mM) than when the calcium concentration was high (1.0 mM); no significant difference was seen in the release of45Ca into the medium at different calcium concentrations. Larger amounts of both isotopes were lost when the prelabeled bones were cultured in resorption-inducing media than in growth-inducing media. When parathyroid hormone stimulated bone resorption in a resorption-inducing medium, equal proportions of both isotopes and bone collagen were lost. Greater losses of3H-tetracycline than of45Ca suggest that45Ca was conserved locally during the resorption that accompanies bone growth, but not during resorption that accompanies bone atrophy.
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Baylink, D., Stauffer, M., Wergedahl, J., Rich, C.: Formation, mineralization, and resorption of bone in vitamin D-deficient rats. J. Clin. Invest.49, 1122–1134 (1970)
Bennett, J.V., Mickelwait, J.S., Barrett, J.E., Brodie, J.C., Kirby, W.M.: Comparative serum bindings of four tetracyclines under simulated in vivo conditions. Antimicrob. Agents Chemotherap.5, 180–182 (1965)
Bingham, P.J., Raisz, L.G.: Bone growth in organ culture: Effects of phosphate and other nutrients on bone and cartilage. Calcif. Tiss. Res.14, 31–48 (1974)
Chen, T.L., Raisz, L.G.: The effects of ascorbic acid deficiency on calcium and collagen metabolism in cultured fetal rat bones. Calcif. Tiss. Res.17, 113–127 (1975)
Dietrich, J.W., Canalis, E.M., Maina, D.M., Raisz, L.G.: Hormonal control of bone collagen synthesis in vitro: Effects of parathyroid hormone and calcitonin. Endocrinology98, 943–949 (1976)
Frost, H.M.: Tetracyclines and fetal bone. Henry Ford Hosp. Med. Bull.13, 403–410 (1965)
Gaillard, P.J.: Parathyroid and bone in tissue culture. In: The parathyroids (Greep, R.O., Talmage, R.V., eds.), pp. 20–45. Springfield, Ill.: Thomas 1961
Gassner, E., Sayegh, F.S.: The nature of tetracycline complex in hard tissue of bovine teeth. Arch. Oral Biol.13, 597–599 (1968)
Ghosez, J.P.: La microscopie de fluorescence dans l'étude du remaniement haversien. Arch. Biol. (Liège)70, 169–178 (1959)
Goldhaber, P., Rabadjija, L., Beyer, W.R., Kornhauser, A.: Bone resorption in tissue culture and its relevance to periodontal diseases. J. Am. Dent. Assn.87, 1027–1033 (1973)
Gruden, N.: Calcium metabolism in the rat in relation to prolonged administration of tetracycline. Calcif. Tiss. Res.13, 41–46 (1973)
Harris, W.H., Jackson, R.H., Jowsey, J.: The in vivo distribution of tetracycline in canine bone. J. Bone Joint Surg.44A, 1308–1320 (1962)
Hevesy, G.: Conservation of skeletal calcium atoms through life. Kgl. Danske Vid. Selskab. Biol. Medd.22, 1–23 (1955)
Ibsen, K.H., Urist, M.R.: The biochemistry and physiology of the tetracyclines: With special reference to mineralized tissues. Clin. Orthopaed.32, 143–168 (1964)
Jackman, K.V., Klein, L., Lacey, S.H.: Recovery and determination of3H-tetracycline from whole bones. Biochem. Med.8, 114–122 (1973)
Jeffay, H., Bayne, H.R.: Bone resorption and45Ca turnover in growing rats. Am. J. Physiol.200, 335–340 (1961)
Kaitila, I.: Effect of tetracycline on mineralization in cycloheximidetreated bones in vitro. Calcif. Tiss. Res.7, 46–57 (1971)
Kaitila, I., Wartiovaara, J., Laitinen, O., Saxen, L.: The inhibitory effect of tetracycline on osteogenesis in organ culture. J. Embryol. Exp. Morph.23, 185–211 (1970)
Kelly, R.G., Buyske, D.A.: Metabolism of tetracycline in the rat and the dog. J. Pharmacol. Exp. Therap.130, 144–149 (1960)
Klein, D.C., Raisz, L.G.: Role of adenosine-3′,5′-monophosphate in the hormonal regulation of bone resorption: Studies with cultured fetal bone. Endocrinology89, 818–826 (1971)
Klein, L.: The action of collagenase on native collagen. Ph.D. thesis, Boston University 1958
Klein, L.: Hydroxyproline in urine and tissues. Std. Methods Clin. Chem.6, 41–56 (1970)
Klein, L., Jackman, K.V.: Quantitative studies on whole bone resorption in vivo: Utilizing3H-tetracycline and3H-proline. In: Proceedings, Ninth European Symposium on Calcified Tissues (Czitober, H., Eschberger, J., eds.), pp. 327–331. Vienna: Facta-Publication 1973
Klein, L., Jackman, K.V.: Assay of bone resorption in vivo with3H-tetracycline. Calcif. Tiss. Res.20, 275–290 (1976)
Klein, L., Lunseth, P.A., Lacey, S.H.: Conservation of skeletal collagen during growth remodeling of bone. Fed. Proc.30, 346, #863 (1971)
Klein, L., Reilly, D.T.: Concurrent exchange of45Ca and3H-tetracycline from rat bone in vitro. Calcif. Tiss. Res.20, 229–234 (1976)
Likins, R.C., Pakis, G.A.: Bone growth and uptake of radiocalcium in tetracycline-treated rats. Nature203, 1069–1070 (1964)
Likins, R.C., Scow, R.D., Zipkin, I., Steere, A.C.: Deposition and retention of fluoride and radiocalcium in the growing rat. Am. J. Physiol.197, 75–80 (1959)
Mandl, I.: Collagenases and elastases. Adv. Enzymol.23, 163–264 (1961)
Marshall, J.H., Rowland, R.E., Jowsey, J.: Microscopic metabolism calcium in bone. V. The paradox of diffuse activity and longterm exchange. Radiation Res.10, 258–270 (1959)
Neuman, W.F., Bareham, B.J.: Evidence for the presence of second calcium phosphate in bone and its stabilization by acid production. Calcif. Tiss. Res.18, 161–172 (1975)
Perrin, D.D.: Binding of tetracyclines to bone. Nature208, 787–788 (1965)
Pickering, D.E., Foran, R.F., Scott, K.G., Crane, J.T.: Chemical growth dynamics of the skeleton in the immature rat. I. Normal chemical composition and growth. Am. J. Dis. Child.92, 276–283 (1956)
Powis, G.: A study of the interaction of tetracycline with human serum lipoproteins and albumin. J. Pharm. Pharmacol.26, 113–118 (1974)
Raisz, L.G.: Bone resorption in tissue culture. Factors influencing the response to parathyroid hormone. J. Clin. Invest.44, 103–116 (1965)
Raisz, L.G., Niemann, I.: Effect of phosphate, calcium and magnesium on bone resorption and hormonal responses in tissue culture. Endocrinology85, 446–452 (1969)
Raisz, L.G., Trummel, C.L.: Effect of vitamin D metabolites on bone resorption in tissue culture. In: Endocrinology, 1971, Proceedings, Third International Symposium (Taylor, S., ed.), pp. 480–486. London: Heinemann Ltd. 1972
Ramp, W.K., Neuman, W.F.: Some factors affecting mineralization of bone in tissue culture. Am. J. Physiol.220, 270–274 (1971)
Stewart, D.J.: The re-incorporation in calcified tissues of tetracycline released following its deposition in the bone of rats. Arch. Oral Biol.18, 759–764 (1973)
Walser, M.: Ion association. VI. Interactions between calcium, magnesium, inorganic phosphate, citrate and protein in normal human plasma. J. Clin. Invest.40, 723–730 (1960)
Ziv, G., Sulman, F.G.: Binding of antibiotics to bovine and ovine serum. Antimicrobiol. Agents Chemotherap.2, 206–213 (1972)
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Chen, T.L., Klein, L. Fetal rat bone in organ culture: Effect of bone growth and bone atrophy on the comparative losses of45Ca and3H-tetracycline. Calc. Tis Res. 25, 255–263 (1978). https://doi.org/10.1007/BF02010779
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DOI: https://doi.org/10.1007/BF02010779