Biomedical subjects
Publications and source records attributed to K Josephsen.
Developing permanent teeth of different ages were obtained from Danish Landrace pigs. Visibly distinct zones on their enamel surfaces were shown to correspond to changes in chemical composition previously reported for other species. The time of appearance, rate of progress and duration of each stage was determined for each tooth type.
The aim of this study was to test whether dental fluorosis can be produced by administration of chronic doses of fluoride during only the post-secretory stage of enamel mineralization. Eight control and eight experimental pigs matched by weight and litter were fed a low-fluoride diet (less than 0.05 mg F-/kg b.w. daily) from weaning to slaughter at 14 months. The test group received an oral dose of 2 mg F-/kg b.w. per day from 8 months of age. Lower fourth pre-molars were at the post-secretory stage at the start of fluoride administration (confirmed by tetracycline marker) and were just erupting at slaughter. All of the fourth pre-molar teeth from the test group developed diffuse enamel hypomineralization indistinguishable from human fluorosis. No such lesions were seen in any of the teeth from the control animals. It was concluded that enamel fluorosis may be caused by fluoride exposure in the maturation phase only. The pathogenic mechanism may be an effect either on the selective loss of protein or on the influx of mineral, both of which occur during the post-secretory or maturation stage of enamel formation.
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Rats were injected with Calcein and killed 1 h, and 1, 2 and 3 days after injection. Incisors were stained with glyoxal-bis-(2-hydroxyanil) (GBHA) and the enamel surface examined in a fluorescence microscope with incident-light excitation. In maturing enamel Calcein produced five fluorescent double bands across the long axis of the tooth. GBHA produced a series of single red stripes, each of which was imposed on a double Calcein band 1 h after injection. At longer intervals after injection GBHA stripes were seen to be gradually displaced in the apical direction from the Calcein double bands. Since GBHA is known to stain enamel adjacent to smooth-ended ameloblasts this supports the view of a cyclic modulation of ameloblast morphology and function.
After injection of labeled precursors such as 3H-proline or 3H-tyrosine into rats, the incisor dentin contains a continuous and stable record of precursor incorporation into labeled proteins. This record was visualized and quantitated with radioautography in order to evaluate the quantitative changes in enamel where newly secreted proteins randomize with older proteins and both are eventually lost. Up to 4 hours after injection, the pulse-dose was incorporated as a highly labeled band of predentin. The band was entirely within calcified dentin at 2 days and was further removed from new predentin by 4 and 8 days. Dentin which formed proximal to the heavily labeled band contained an amount of radioactivity reflecting the level of labeled precursor available at that time. A standardizing factor for experimental error was obtained by quantitating the reaction in the heavily labeled band, and a post-pulse incorporation factor was determined from the amount of radioactivity added per day as weakly labeled dentin. The variation within the heavily labeled band was assumed to reflect experimental error. The number of grains in the bands were averaged from 4 hours to 8 days to give the standardizing factor. This was multiplied by the ratio of enamel to dentin counts in the same section to obtain a corrected enamel count. In this way the coefficient of variation was improved from a high of 17.2% in uncorrected enamel counts to 2.4% in corrected counts. The post-pulse incorporation factor was higher with tyrosine than with proline. With proline it amounted to 5% increase per day from 1 to 4 days and 2.5% per day from 4 to 8 days after injection. In addition, with 3H-proline the incorporation into predentin increased from 30 minutes to 4 hours. With tyrosine, the counts increased from 30 minutes to 1 hour, but decreased by nearly one third from 1 to 4 hours. This was interpreted as a loss of short-lived matrix proteins including procollagen peptides produced during conversion from procollagen to tropocollagen in the predentin.
Odontoblasts are cells with single cytoplasmic processes that grow longer as more dentin is elaborated. Ameloblasts also have single processes and it has been postulated that they too grow longer as more enamel is made. Support for this hypothesis was obtained using rat incisors to investigate the behavior of substances labeled with 3H-proline and 3H-fucose. A comparison was made between odontoblasts, which have processes known to grow and remain within the dentin, and the ameloblasts whose Tomes' processes are hypothesized to grow and leave remnants in the completed enamel. With 3H-proline, the odontoblast bodies are labeled at the early time intervals. They synthesize and secrete a layer of intensely labeled predentin, which by 1 and 2 days is converted to mineralized dentin. Matrix deposited after the main pulse is weakly labeled. Odontoblast processes are never labeled in dentin formed prior to injection. With 3H-fucose, the cell bodies are labeled at the early intervals and the newly formed glycoproteins are deposited into the predentin. Almost immediately, these are progressively added to the dentin at the calcification front. With time a gradient of labeling extends from the unlabeled dentin toward the odontoblast bodies. Unlike the behavior of labeled proteins, by 1 and 2 days labeled glycoproteins appear along the entire length of the odontoblast processes. In the enamel, no Tomes' processes are present during maturation. With 3H-proline, reactions are adjacent to the cells and diffuse toward, but do not reach the dentino-enamel junction by 1 and 2 days. With 3H-fucose, reactions appear over the enamel near the cells. By 1 and 2 days no diffusive pattern is seen, but grains are concentrated near the dentino-enamel junction, in a region containing holes known to be the beginning of Tomes' processes. Since odontoblast glycoproteins migrate along odontoblast processes, it was postulated that cytoplasmic remnants were present in enamel along which ameloblast glycoproteins could also migrate to reach the holes at the dentino-enamel junction.
The rat incisor is an excellent model system in which to study amelgenesis. However, the information obtained has not been extrapolated to the human because of alleged structural differences between the teeth. The obvious differences include continuous eruption in rat incisors and an enamel rod pattern in rats which seemingly differs from the keyhole pattern of human enamel. A comprehensive analysis was made of those features of enamel structure considered fundamental to the understanding of its formation. This was done by applying the knowledge of amelogenesis obtained in rat incisors to the teeth of monkey and man. The following points of basic similarity were established between these species: (1) Interrod enamel is secreted first. It forms the side walls of cavities which are initially occupied by Tomes' processes. (2) The formation of interrod cavities is followed by deposition of enamel rods within these spaces. (3) The rods conform to the shape of the cavities and are secreted from one surface of Tomes' process. (4) At the initial site of rod deposition its enamel is continuous with the interrod enamel wall. (5) Growth of the rod compresses the process to one side of the cavity resulting in an arcade-shaped "space" between the rod and the remaining interrod walls. This study demonstrates that it is no longer necessary to postulate a keyhole structure for primate enamel, and it has established that a fundamental similarity exists in the basic structure and in the mode of formation of enamel in all three species.
The ultrastructural features of the epithelial-mesenchymal interface in a case of an ameloblastic fibro-odontoma was studied with special reference to possible signs of "inductive" processes. In most parts of the tumor, the odontogenic epithelium was separated from the connective tissue by a thick rim of a finely filamentous meshwork in which a basal lamina was occasionally observed. Mesenchymal cells were seen to touch the filamentous meshwork but no membrane bound matrix vesicles were recorded. Small areas of dentin-like tissue were found in the juxtaepithelial connective tissue while enamel-like areas and spherical calcified masses were encountered in epithelial islands. The organic matrix in relation to the enamel-like tissue consisted of either tubular fibers or a fine-granular material. It was assumed that the tubular matrix component directed the formation of long enamel-like crystals, and that the fine-granular matrix was degraded tubular fibers in which spherical calcified masses might arise. Spherical calcified masses could be found in separate follicles also where they were related to a fine-fibrillar matrix or collagenous material. The cell layers forming the wall of the islands had a great resemblance to those of an enamel organ, but the findings of dentin-free, enamel-like areas are not compatible with the inductive theory of normal odontogenesis.
Three groups of 1-month-old rats were kept on water containing 0.2, 56.5 and 113 parts/10(6) F-, respectively, for a period of 2 months. These groups were set up in order to study the relationship between long-term administration of high doses of waterborne fluoride, plasma values of fluoride and calcium and changes in forming dentin and enamel of the rat incisor. The microradiographic study revealed that increased levels of fluoride caused a delay in mineralization of the enamel. In the 113 parts/10(6) group the radiolucent areas in the enamel exhibited a distinct periodicity along the tooth, a phenomenon which cannot at present be explained. In the dentin the fluoride interfered with the process of mineralization giving rise to radiolucent and radiopaque banding in the experimental groups. With increasing levels of fluoride in the water supply a significant increase of plasma fluoride was recorded. In contrast the serum calcium values in both experimental groups were reduced to the same extent, which differed significantly from that of the control group. It is concluded that the fluoride doses which are needed to create fluorotic changes in rats may interfere with calcium metabolism and basic mineralization processes.
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