Summary
Staining reactions for a number of histochemical procedures for lipophilic staining and protein were studied in the enamel matrix along the length of rat incisors. Sudan Black gave a positive stain across the whole thickness of very early enamel (up to 30 μm) but this staining only continued as a narrow band close to the ameloblasts as the enamel matured. A variety of tests for protein produced almost identical staining patterns in enamel matrix up to 100 μm thick. Since the pattern of “lipid” staining persisted, after using a number of procedures which could normally be expected to remove lipid, it is suggested that Sudan Black positive staining may be due to lipophilic protein rather than lipid itself. Fluoride did not significantly alter the staining reactions for “lipid” and protein but did proceduce matrix which was much more effectively stained by cross-linking agents FFDNB and FF sulphene.
Similar content being viewed by others
References
Adkins, B.L., Kruger, B.J.: Statistical evaluation of a multiple response experiment: Alterations to the morphology of rat molars. J. dent. Res.45, 1205–1213 (1966)
Basford, K.E., Patterson, C.M., Kruger, B.J.: Multivariate analyses of the influence of mottling doses of fluoride on the amino acids of enamel matrix protein from rat incisors. Archs. oral Biol.21, 121–129 (1976)
Boyde, A.: In Second International Symposium on the Composition, Properties and Fundamental Structure of Tooth Enamel (M.V. Stack, R.W. Fearnhead, eds.), p. 133, Bristol: John Wright 1971
Brain, E.B.: Rapid demineralization for microscopy of tooth enamel and associated structures. Br. dent. Jnl.123, 177–181 (1967)
Deutsch, D., Weatherell, D., Robinson, C.: Distribution of fluoride in developing bovine enamel. J. dent. Res.53, 1053 (1974).
Eastoe, J.E., Starkey, W.E.: Demonstration of local changes in developing enamel by differential staining. J. dent. Res.46, 1285 (1967)
Eastoe, J.E., Camilleri, G.E.: The pattern of protein distribution in enamel during maturation. In: Second International Symposium on the Composition, Properties and Fundamental Structure of Tooth Enamel (M.V. Stack, R.W. Fearnhead, eds.), pp. 119–123. Bristol: John Wright 1971
Fincham, A.G.: Experiments on the D.E.A.E.-cellulose chromatography of bovine foetal enamel matrix and the isolation of low molecular weight fractions. In: Second International Symposium on the Composition, Properties and Fundamental Structure of Tooth Enamel (M.V. Stack, R.W. Fearnhead, eds.), pp. 79–87. Bristol: John Wright 1971
Goland, P., Scheiman-Tagger, E., Engel, M.: Enamel preservation during decalcification following fixation by some reactive halogen compounds. J. dent. Res.44, 342–349 (1965)
Hallsworth, A.S., Weatherell, J.A., Deutsch, D.: An ultrasensitive method of determining fluoride ion in mineralised tissues. J. dent. Res.53, 1088 (1974)
Irving, J.T., Wuthier, R.E.: Further observations on the sudan black stain for calcification. Arch. oral Biol.5, 323–324 (1961)
Irving, J.T.: Histochemical changes in the early stages of calcification. Clin. Orth. and Rel. Res.17, 92–101 (1960)
Irving, J.T.: The sudanophil material at sites of calcification. Arch. oral Biol.8, 735–745 (1963)
Irving, J.T., Wuthier, R.E.: Histochemistry and biochemistry of calcification with special reference to the role of lipids. Clin. Orth. and Rel. Res.56, 237–260 (1968)
Irving, J.T.: The pattern of sudanophilia in developing rat molar enamel. Arch. oral Biol.18, 137–140 (1973a)
Irving, J.T.: Theories of mineralization inCalcium and Phosphorus Metabolism, pp. 81–92. New York: Academic Press 1973b
Kruger, B.J.: Histological effects of fluoride and molybdenum on developing dental tissues. Aust. dent. J.12, 54–60 (1967)
Kruger, B.J.: Ultrastructural changes in ameloblasts from fluoride treated rats. Arch. oral Biol.13, 969–977 (1968)
Kruger, B.J.: The effect of different levels of fluoride on the ultrastructure of ameloblasts in the rat. Arch. oral Biol.15, 109–114 (1970)
Lillie, R.D.: Histopathologic technic and practical histochemistry, 3rd ed., pp. 561–565. New York: McGraw-Hill 1965
Lowenstein, J.M.: In: Lowenstein, J.M. (ed.), Methods in enzymology, pp. 131–213. New York: Academic Press 1969
Mahler, H.R., Cordes, E.H.: Biological Chemistry, pp. 70–120. New York: Harper 1966
McManus, J.F.A., Mowry, R.W.: Staining methods. Histologic and histochemical, p. 100. New York: Harper & Row 1964
Melcher, A.H.: Extracellular sudanophilic material in dental structures. Arch. oral Biol.12, 913–916 (1967).
Patterson, C.M., Basford, K., Kruger, B.J.: The effect of fluoride on the immature enamel matrix protein of the rat. Arch. oral Biol. (in press 1977)
Pearse, A.G.E.: Histochemistry 3rd ed., pp. 424–425, 694–695 London: Churchill 1968
Robinson, C., Fuchs, P., Weatherell, J.A.: Changes in the amino acid composition of developing cow enamel. J. dent. Res.53, 1054 (1974)
Shapiro, I.M., Wuthier, R.E., Irving, J.T.: A study of the phospholipids of bovine dental tissues — I. Arch. oral Biol.11, 501–512 (1966)
Thompson, S.W.: Selected histochemical and histopathological methods, pp. 332–333. Springfield, Ill.: Thomas 1966
Walton, R.E., Eisenmann, D.R.: Ultrastructural examination of various stages of amelogenesis in the rat following parenteral fluoride administration. Arch. oral Biol.19, 171–182 (1974)
Wells, M.A., Hanahan, D.J.: Studies on phospholipase. 1. Isolation and characterization of two enzymes fromcrotalus adamanteus venom. Biochemistry8, 414–421 (1969)
Zerlotti, E., Yaeger, J.A.: Histochemistry and biophysical histology of the matrices of some mineralized tissues. Clin. Orth. and Rel. Res.51, 223–253 (1967)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Patterson, C.M., Kruger, B.J. & Daley, T.J. Lipid and protein histochemistry of enamel — Effects of fluoride. Calc. Tis Res. 24, 119–125 (1977). https://doi.org/10.1007/BF02223305
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02223305