The effect of a single dose of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) on secretory ameloblasts and enamel formation in rat incisors.

The present experiment was undertaken in order to study how HEBP affects secretory ameloblasts and the mineralizing front of rat incisor enamel resulting in the formation of a hypomineralized incremental band. Rats were given a single subcutaneous injection of 10 mg/kg body weight of HEBP and sacrificed at intervals ranging from 1 hour to 9 days. The incisors were specially prepared for microradiography, scanning electron microscopy, and light and transmission electron microscopy. The microradiographical examination 9 days after injection revealed a distinct incremental band of hypomineralization from the amelo-dentinal junction to the outer enamel surface. The overall rod pattern and mineral distribution within the enamel was otherwise normal. Under light microscopy the ameloblasts exhibited an increase in vacuoles and dark granules in the supranuclear cytoplasm 2-12 hours after injection. At 12 hours a total disarray of the mineralizing front was evident and confirmed by SEM. However, 24 hours after injection a normal structure of the mineralizing front was regained. Ultrastructurally the organelles of the ameloblasts showed no changes from normal at any time interval. However, 2 hours after injection multivesicular bodies appeared frequently in the Tomes' processes and crystal density had diminished in the interrod enamel. At 8 hours a proteinaceous matrixdevoid of crystals was accumulating corresponding to interrod growth regions, whereas no obvious changes were recorded at the rod growth regions. At 12 hours the newly formed interrod enamel was thin and without crystals in some places. In such areas a granular, less-dense matrix was found and a band of small swellings of the interrod enamel was evident most likely corresponding to the time of injection. After 24 hours the cells and the mineralizing front appeared normal except in the interrod growth regions where the cell processes were still separated by wide intercellular spaces in which crystals were occasionally found in a fine granular matrix. These observations are discussed with particular reference on the known physico-chemical effects of HEBP on mineral formation. A possible direct cellular effect on the ameloblasts should be studied using radioautography and immunocytological techniques.