Localization of Cellular Calcium in Differentiating Ameloblasts and its Relationship to the Early Mineralization Process in Mantle Dentin and Enamel in Hamster Tooth Germs in Vitro

¹ Laboratory of Tooth Development, Department of Oral Cell Biology, School of Dentistry ACTA, Vrije Universiteit, de Boelelaan 1115, 1081 HV Amsterdam, The Netherlands

The relationship between the distribution of calcium in the cells of the enamel organ and the mineralization process in mantle dentin and enamel was investigated at the ultrastructural level in cultured hamster second maxillary molar tooth germs explanted before the onset of mineralization (bell stage). During the early stages of pre-odontoblast and pre-ameloblast differentiation, pyroantimcnate (PA) reaction product for calcium was observed only in the nuclei. However, an abrupt increase in PA reaction product appeared in the apical cytoplasm of both the pre-odontoblasts and pre-ameloblasts prior to the onset of mantle dentin mineralization. In the pre-dentin, the PA reaction product was localized mainly on the striated collagen fibers.The PA reaction product in the apical poles of these cells increased concomitantly with increasing mantle dentin mineralization. The amounts of PA reaction product along the plasma membranes and in the cytoplasm decreased in the direction of the basal (stratum intermedium) pole. The highest PA activity in the cytoplasmic side of the plasma membranes of the ameloblasts was found during the secretory phase of amelogenesis. However, in the area around the tips of the Tomes' processes, membrane-associated and cytoplasmic PA activity was low or absent but gradually increased toward the ameloblast cell body, an indication of the presence of a calcium gradient in the processes.

These results indicate that in vitro: (1) both odontoblasts and (pre)-ameloblasts are involved in the calcium acquisition necessary for the initial stages of mantle dentin mineralization; (2) in ameloblasts, there is a calcium gradient in the direction of the mineralization front from the earliest stages of mantle dentin mineralization onward; (3) enamel matrix does not seem to be involved in calcium translocation to the enamel mineralization front; (4) the Tomes' processes seem to regulate transmembrane calcium transport to the mineralization front; and (5) the distribution of calcium in the enamel organ is comparable with that found in vivo.