High-resolution electron-microscopic study of the relationship between human enamel and dentin crystals at the dentinoenamel junction.

The development of dentin and of enamel share a common starting locus: the dentinoenamel junction (DEJ). In this study the relationship between enamel and dentin crystals has been investigated in order to highlight the guiding or modulating role of the previously mineralized dentin layer during enamel formation. Observations were made with a high-resolution electron microscope and, after digitalization, image-analysis software was used to obtain digital diffractograms of individual crystals. In general no direct epitaxial growth of enamel crystals onto dentin crystals could be demonstrated. The absence of direct contact between the two kinds of crystals and the presence of amorphous areas within enamel particles at the junction with dentin crystals were always noted. Only in a few cases was the relationship between enamel and dentin crystals observed, which suggested a preorganization of the enamel matrix influenced by the dentin surface structure. This could be explained either by the existence of a proteinaceous continuum between enamel and dentin or by the orientation of enamel proteins by dentin crystals.

Initial aspects of mineralization at the dentino-enamel junction in embryonic mouse incisor in vivo and in vitro: a tem comparative study.

The frontier between the enamel organ and the dental papilla, the future dentino-enamel junction, undergoes coordinated modifications. The mineralization of the extracellular matrix starts within the predentine, which is a prerequisite for the formation of the first enamel crystallites in vivo. We investigated the dentino-enamel junction using the embryonic mouse incisor as a model. Our data showed that the notion of the dentino-enamel junction should not be restricted to the thin interface classically described. A temporo-spatial survey from the epithelio-mesenchymal junction to the dentino-enamel junction delineated a clear sequence of events characterized by the early deposition of electron-dense granules, followed by the appearance of patches of stippled material at the dentino-enamel junction. The first tiny enamel crystallites appeared in the vicinity of this material which presented a well-ordered alignment. The comparison of data obtained in vivo on 17-, 18-, 19-d-old embryonic incisors with those obtained in vitro using 15-d-old embryonic incisors cultured for 7 d emphasizes the relevance of this sequence. Helicoidal growing crystals were observed in cultured tooth germs but never in vivo.

First experimental evidence for human dentine crystal formation involving conversion of octacalcium phosphate to hydroxyapatite.

Biological apatite-crystal formation is a complex process starting with heterogeneous nucleation of inorganic calcium phosphate on an organic extracellular matrix [Cuisinier et al. (1995), J. Cryst. Growth, 156, 443-453]. Further stages of crystal growth are also controlled by the organic matrix and both nucleation and growth processes are under cellular control [Mann (1993), Nature (London), 367, 499-505]. The final mineral in calcified tissue is constituted by poorly crystalline hydroxyapatite (HA) with a low Ca:P ratio, containing foreign ions such as carbonate and fluoride. This study reports the first observation of octacalcium phosphate (OCP) [Brown (1962), Nature (London), 196, 1048-1055] in a biological tissue; OCP was found in the central part and HA at the extremities of the same crystal of calcifying dentine. This observation is of key importance in understanding the first nucleation steps of biological mineralization. The presence of OCP in a forming human dentine crystal and the observation in the same tissue of nanometer-sized particles with a HA structure [Houllé et al. (1997), J. Dent. Res. 76, 895-904] clearly proves that two mechanisms, direct nucleation of non-stoichiometric HA crystals and nucleation of OCP, occur simultaneously in same area of mineralization. OCP is found to be a transient phase during the growth of biological crystals. In small crystals, OCP is completely transformed into HA by a hydrolysis reaction (Brown, 1962) and can only be detected in larger crystals because of its slow kinetics of transformation.

High resolution electron microscopy: structure and growth mechanisms of human dentin crystals.

Biological crystal formation was postulated to begin by a nucleation process. Such processes have been demonstrated for human amelogenesis and bone mineralization. The aim of this study was to confirm if such mechanisms occur during dentin crystal formation. The structure of human fetal dentin crystals and the earliest stages of mineral growth were followed by High Resolution Electron Microscopy (HREM) associated with digitalized image analysis. Micrographs of the mineralization front were first digitalized, and selected areas were transformed in the reciprocal space by Fast Fourier Transform. The resulting diffractograms were compared with computer-simulated diffractograms and used to determine the orientation of crystals. Dentin crystals, found close to the mineralization front, show a structure closely related to that of hydroxyapatite (HA), as determined by comparison of HREM images with simulated images. These crystals present numerous structural defects such as dislocations and grain boundaries. These defects appear to be present in dentin crystals at an early stage of growth. We have also observed nanometer-sized particles in mineralization areas. Calculated diffractograms of these areas show significant similarities with HA diffraction patterns, and in one case, their structure could be correlated to HA structure through an image simulation process. These nanometer-sized particles could be related to the nucleation process, and their growth, orientation, and formation appear to be mediated by extracellular matrix components.