O. Fejerskov

Dental Fluorosis: Chemistry and Biology

This review aims at discussing the pathogenesis of enamel fluorosis in relation to a putative linkage among ameloblastic activities, secreted enamel matrix proteins and multiple proteases, growing enamel crystals, and fluid composition, including calcium and fluoride ions. Fluoride is the most important caries-preventive agent in dentistry. In the last two decades, increasing fluoride exposure in various forms and vehicles is most likely the explanation for an increase in the prevalence of mild-to-moderate forms of dental fluorosis in many communities, not the least in those in which controlled water fluoridation has been established. The effects of fluoride on enamel formation causing dental fluorosis in man are cumulative, rather than requiring a specific threshold dose, depending on the total fluoride intake from all sources and the duration of fluoride exposure. Enamel mineralization is highly sensitive to free fluoride ions, which uniquely promote the hydrolysis of acidic precursors such as octacalcium phosphate and precipitation of fluoridated apatite crystals. Once fluoride is incorporated into enamel crystals, the ion likely affects the subsequent mineralization process by reducing the solubility of the mineral and thereby modulating the ionic composition in the fluid surrounding the mineral. In the light of evidence obtained in human and animal studies, it is now most likely that enamel hypomineralization in fluorotic teeth is due predominantly to the aberrant effects of excess fluoride on the rates at which matrix proteins break down and/or the rates at which the by-products from this degradation are withdrawn from the maturing enamel. Any interference with enamel matrix removal could yield retarding effects on the accompanying crystal growth through the maturation stages, resulting in different magnitudes of enamel porosity at the time of tooth eruption. Currently, there is no direct proof that fluoride at micromolar levels affects proliferation and differentiation of enamel organ cells. Fluoride does not seem to affect the production and secretion of enamel matrix proteins and proteases within the dose range causing dental fluorosis in man. Most likely, the fluoride uptake interferes, indirectly, with the protease activities by decreasing free Ca(2+) concentration in the mineralizing milieu. The Ca(2+)-mediated regulation of protease activities is consistent with the in situ observations that (a) enzymatic cleavages of the amelogenins take place only at slow rates through the secretory phase with the limited calcium transport and that, (b) under normal amelogenesis, the amelogenin degradation appears to be accelerated during the transitional and early maturation stages with the increased calcium transport. Since the predominant cariostatic effect of fluoride is not due to its uptake by the enamel during tooth development, it is possible to obtain extensive caries reduction without a concomitant risk of dental fluorosis. Further efforts and research are needed to settle the currently uncertain issues, e.g., the incidence, prevalence, and causes of dental or skeletal fluorosis in relation to all sources of fluoride and the appropriate dose levels and timing of fluoride exposure for prevention and control of dental fluorosis and caries.

Dental Tissue Effects of Fluoride

It is now well-established that a linear relationship exists between fluoride dose and enamel fluorosis in human populations. With increasing severity, the subsurface enamel all along the tooth becomes increasingly porous (hypomineralized), and the lesion extends toward the inner enamel. In dentin, hypomineralization results in an enhancement of the incremental lines. After eruption, the more severe forms are subject to extensive mechanical breakdown of the surface. The continuum of fluoride-induced changes can best be classified by the TF index, which reflects, on an ordinal scale, the histopathological features and increases in enamel fluoride concentrations. Human and animal studies have shown that it is possible to develop dental fluorosis by exposure during enamel maturation alone. It is less apparent whether an effect of fluoride on the stage of enamel matrix secretion, alone, is able to produce changes in enamel similar to those described as dental fluorosis in man. The clinical concept of post-eruptive maturation of erupting sound human enamel, resulting in fluoride uptake, most likely reflects subclinical caries. Incorporation of fluoride into enamel is principally possible only as a result of concomitant enamel dissolution (caries lesion development). At higher fluoride concentrations, calcium-fluoride-like material may form, although the formation, identification, and dissolution of this compound are far from resolved. It is concluded that dental fluorosis is a sensitive way of recording past fluoride exposure because, so far, no other agent or condition in man is known to create changes within the dentition similar to those induced by fluoride. Since the predominant cariostatic effect of fluoride is not due to its uptake by the enamel during tooth development, it is possible to obtain extensive caries reductions without a concomitant risk of dental fluorosis.

Detection of occlusal caries without cavitation by visual inspection, film radiographs, xeroradiographs, and digitized radiographs.

The study compared visual inspection, conventional film radiographs, xeroradiographs, and digitized radiographs for the detection of caries in occlusal surfaces without cavitation. 166 extracted premolars and permanent molars without macroscopic cavitation were included. Eight observers assessed five grades of occlusal caries by visual inspection and by the three imaging techniques. Histologic sections (700-1,000 microns) served as validating criterion for the presence and depth of carious lesions, 82 teeth being found carious (27 with deep dentinal involvement) and 84 being scored as sound. Approximately 20% of the dentinal carious lesions were detected by visual inspection and 40% by conventional film radiography (both with a minimal number of false-positive observations). By xero- and digital radiography, an increasing number of false-positive scorings was seen. By digital radiography the true-positive detection rate was similarly increased to 60%. The predictive values of a positive test were, on average, 0.85 for visual inspection, 0.89 for film, 0.78 for xero-, and 0.82 for digital radiography and for a negative test 0.56, 0.61, 0.61, and 0.69, respectively. However, digital radiography was able to detect over 70% of deep dentinal lesions in contrast to 45% by the other two imaging methods, without an increase in false-positive diagnoses of truly caries free teeth, compared with the other techniques.

Dental Fluorosis Developed in Post-secretory Enamel

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 (<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.

Development of Dental Fluorosis according to Age at Start of Fluoride Administration

The aim was to determine at what ages the different types of human permanent teeth are susceptible to development of fluorosis. The prevalence of disturbances of enamel mineralization was recorded blindly in 70 children who had previously participated in a fluoride tablet program (0.5 mg F–– per day) and 40 other children who had never received fluoride tablets. Comparison of the prevalence of enamel lesions between these two groups and between the ages at which children began to take the tablets revealed the ages at which there was an increased risk of fluorosis. The data showed that fluoride can affect teeth late in their development when enamel is in a stage of late secretion or early maturation. It was concluded that this finding, which is in accordance with experimental animal studies, calls for a fundamental change in thinking not only with respect to the mechanisms involved in human enamel fluorosis, but also in relation to timing of fluoride programs.

Tooth Mortality in an Adult Rural Population in Kenya

This paper reports on the pattern of tooth loss in a random sample of 1131 adults aged from 15 to 65 years in a rural area of Kenya in which access to formal dental care is minimal. We found that the majority of the population retained most of their dentition in a functional state even up to the age of 65 years: In all age groups, more than 50% had at least 26 teeth present, and more than 90% had at least 16 teeth present. The prevalence of edentulousness was less than 0.3%. The principal cause of tooth loss in all age groups was caries, and this was true for all tooth-types except incisors, for which periodontal disease was the main cause of tooth loss. The cultural practice of removing lower central incisors was observed only in those over 40 years of age. More teeth were lost due to caries among women than among men, while the reverse was true for teeth lost due to periodontal diseases. In view of the fact that most people retain most of their teeth throughout life, it is suggested that the most appropriate strategies for dental health care in this population should be those promoting self care, rather than the introduction of a formal treatment-oriented approach provided by dentists.

Ion transporters in secretory and cyclically modulating ameloblasts: a new hypothesis for cellular control of preeruptive enamel maturation

Mature enamel consists of densely packed and highly organized large hydroxyapatite crystals. The molecular machinery responsible for the formation of fully matured enamel is poorly described but appears to involve oscillative pH changes at the enamel surface. We conducted an immunohistochemical investigation of selected transporters and related proteins in the multilayered rat incisor enamel organ. Connexin 43 (Cx-43) is found in papillary cells and ameloblasts, whereas Na(+)-K(+)-ATPase is heavily expressed during maturation in the papillary cell layer only. Given the distribution of Cx-43 channels and Na(+)-K(+)-ATPase, we suggest that ameloblasts and the papillary cell layer act as a functional syncytium. During enamel maturation ameloblasts undergo repetitive cycles of modulation between ruffle-ended (RA) and smooth-ended (SA) ameloblast morphologies. Carbonic anhydrase II and vacuolar H(+)-ATPase are expressed simultaneously at the beginning of the maturation stage in RA cells. The proton pumps are present in the ruffled border of RA and appear to be internalized during the SA stage. Both papillary cells and ameloblasts express plasma membrane acid/base transporters (AE2, NBC, and NHE1). AE2 and NHE1 change position relative to the enamel surface as localization of the tight junctions changes during ameloblast modulation cycles. We suggest that the concerted action of the papillary cell layer and the modulating ameloblasts regulates the enamel microenvironment, resulting in oscillating pH fluctuations. The pH fluctuations at the enamel surface may be required to keep intercrystalline spaces open in the surface layers of the enamel, enabling degraded enamel matrix proteins to be removed while hydroxyapatite crystals grow as a result of influx of calcium and phosphate ions.

Pulp-capping with recombinant human insulin-like growth factor I (rhIGF-I) in rat molars.

The aim of this study was to explore pulp healing and reparative dentinogenesis following pulp-capping by using recombinant human insulin-like growth factor I (rhIGF-I). Exposures were made through the mesial pulp horn in first upper molars in two-month-old Wistar rats. The pulp was covered with one dose of sterile 4% methylcellulose gel containing either 400 ng rhIGF-I or saline in contralateral controls. The exposure site was closed with sterile Teflon membrane, and the cavity was filled with IRM cement. Additional molars were capped with Dycal as controls. After 3, 7, or 28 days, animals were anesthetized and fixed by intravascular glutaraldehyde perfusion. Molars were decalcified and processsed for histological analysis and cut with membrane and residual methacrylate from IRM in situ. Only specimens with acceptable pulp sealing according to blinded microscopy control were included. On day 3, identical inflammatory responses in the upper pulp were observed in molars with rhIGF-I gel or control gel. On day 7, granulation tissue ingrowth had partly replaced inflammatory infiltration in both groups. After 28 days, complete dentin bridging and tubular dentin formation were observed more frequently and closer to the test substance containing rhIGF-I. The reparative dentin response to capping with rhIGF-I was similar to that after the use of Dycal. In conclusion, microscopic control of membrane sealing in situ gives valid information on the more subtle pulp effects of growth factors. The observations suggest that pulp-capping of rat molars by means of rhIGF-I enhances reparative dentinogenesis in comparison with vehicle controls.

The Effect of Sucrose on Plaque pH in the Primary and Permanent Dentition of Caries-inactive and -active Kenyan Children

The hypothesis that the Stephan pH responses of dental plaque would be different in caries-active and -inactive individuals was tested in 20 seven-year-old and 19 14-year-old Kenyan children. In each age group, half the children had ≥ 2 dentin cavities; the other half had no such lesions. With a palladium-touch microelectrode, interdental plaque pH was monitored between m1/m2 in each quadrant in the primary dentition and in the four molar/premolar regions in the permanent dentition. pH was also monitored in caries cavities in the occlusal surfaces of lower first molars and on the tongue. pH was measured before and up to 60 min after the children rinsed with 10 mL of 10% sucrose. Caries status of the individual was unrelated to plaque pH in comparable non-carious sites in both of the age groups. The pH minimum in the maxilla was about 0.5 pH units lower than that in the mandible. Active occlusal caries lesions had a resting pH value of about 5.5, about 1 pH unit lower than that of sound surfaces. The pH dropped to about 4.5 in caries lesions and recovered slowly. In sound occlusal sites, a pH drop to about 6.0 was followed by a relatively rapid return to the resting value. Thus, when the mean values were considered, the classic Stephan curve response was evident. However, when the pH changes at single sites were considered at various time intervals, a substantial, erratic fluctuation was observed. The tongue had a resting pH value of 7.0, which dropped to about pH 5.5 and remained low for more than 60 min.

Arrest of Root Surface Caries in situ

This study tests the hypothesis that daily oral hygiene combined with topical fluoride arrests active root-surface caries lesions without changing the mineral content of the lesions. Therefore, changes in mineral content and distribution were studied in root surfaces during caries lesion development and subsequent arrest of lesion progression in situ. In 18 subjects, lesions were developed during 3 months in sound root-surface specimens inserted into lower partial dentures. After 3 months, ground sections were prepared from each lesion prior to re-insertion of the specimens into the dentures. In addition, one sound root specimen was added per subject. During the following 3 months, half of the subjects cleaned both sound and carious specimens once a day with an 1100-ppm fluoride toothpaste, and the specimens were treated twice with 2% NaF for 2 min in situ. The other half of the subjects continued the experiment without cleaning. During the initial three-month period, all specimens developed subsurface lesions extending 187 to 583 μm into the dentin. Lesion depth increased somewhat in both experimental groups during the following 3 months (P ≥ 0.1). There was a non-significant increase in mineral loss in the plaque-covered specimens (P = 0.08). However, the total mineral content of specimens subjected to plaque removal and topical fluoride did not change. This treatment resulted in an increased mineral content in the surface layer (P < 0.01) and formation of a zone of higher mineral content within the body of the lesion. The sound root surfaces which had been cleaned for a three-month period showed mineral uptake in the surface layer, occasionally associated with subsurface demineralization extending 20 to 70 μm into the tissue. The mineral loss of these specimens was significantly smaller than that of plaque-covered surfaces (P < 0.001). It is concluded that daily plaque removal and topical fluoride use influence the distribution of mineral in sound and carious root surfaces and may arrest lesion progression without affecting the total mineral content.