Multi-scale characterization of Developmental Defects of Enamel and their clinical significance for diagnosis and treatment.

Developmental Defects of Enamel (DDE) such as Dental Fluorosis (DF) and Molar Incisor Hypomineralization (MIH) are a major public health problem. Their clinical aspects are extremely variable, challenging their early and specific diagnosis and hindering progresses in restorative treatments. Here, a combination of macro-, micro- and nano-scale structural and chemical methods, including, among others, Atom Probe Tomography recently applied on tooth enamel, were used to study and compare MIH, DF and healthy teeth from 89 patients. Globally, we show that DF is characterized by an homogenous loss of mineral content and crystallinity mainly disrupting outside layer of enamel, whereas MIH is associated with localized defects in the depth of enamel where crystalline mineral particles are embedded in an organic phase. Only minor differences in elemental composition of the mineral phase could be detected at the nanoscale such as increased F and Fe content in both severe DDE. We demonstrate that an improved digital color measurement of clinical relevance can discriminate between DF and MIH lesions, both in mild and severe forms. Such discriminating ability was discussed in the light of enamel composition and structure, especially its microstructure, organics presence and metal content (Fe, Zn). Our results offer additional insights on DDE characterization and pathogenesis, highlight the potentiality of colorimetric measurements in their clinical diagnosis and provide leads to improve the performance of minimally invasive restorative strategies. STATEMENT OF SIGNIFICANCE: Developmental Defects of Enamel (DDE) are associated to caries and tooth loose affecting billions of people worldwide. Their precise characterization for adapted minimally invasive care with optimized materials is highly expected. Here In this study, first we propose the use of color parameters measured by a spectrophotometer as a means of differential clinical diagnosis. Second, we have used state-of-the-art techniques to systematically characterize the structure, chemical composition and mechanical optical properties of dental enamel teeth affected by two major DDE, Dental Fluorosis (DF) or Molar Incisor Hypomineralization (MIH). We evidence specific enamel structural and optical features for DF and MIH while chemical modifications of the mineral nanocrystals were mostly correlated with lesion severity. Our results pave the way of the concept of personalized dentistry. In the light of our results, we propose a new means of clinical diagnosis for an adapted and improved restoration protocol for these patients.

Chronic Exposure to Bisphenol A Exacerbates Dental Fluorosis in Growing Rats.

Enamel defects resulting from environmental conditions and way of life are public health concerns because of their high prevalence. Because their etiology is unclear, the aim of this study was to analyze the various forms of enamel hypomineralization, and to characterize the genes involved in this process to determine the mechanisms involved in disruptions of amelogenesis. We used bisphenol A (BPA) and fluoride as models; both are commonly encountered in human populations and utilized in dentistry. Wistar rats were chronically exposed to 5 µg/kg/day BPA from day 1 of gestation to day 65 after birth (P65) and 5 mM fluoride from P21 to P65. Resulting enamel defects were comparable to the human enamel pathologies molar incisor hypomineralization (MIH) and dental fluorosis (DF) respectively, and were more severe in rats exposed to both agents than to each agent alone. Large-scale transcriptomic analysis of dental epithelium showed a small group of genes the expression of which was affected by exposure to BPA or NaF. Among the most modulated, many are directly involved in amelogenesis (Amelx, Enam, Klk4, Mmp12, Slc26a4, and Slc5a8), and can be regrouped as forming the "hypomineralization enameloma." Each of these gene expression perturbations may contribute to enamel defects. Exposure to BPA weakens enamel, making it more prone to generate frequent mineralization defects MIH and DF. Our study identifies hypomineralization genes that may enable the use of dental enamel as an early marker of exposure to environmental toxicants because of its unique ability to retrospectively record ameloblast pathophysiology. © 2016 American Society for Bone and Mineral Research.

Marine steroids as potential anticancer drug candidates: In silico investigation in search of inhibitors of Bcl-2 and CDK-4/Cyclin D1.

Star fishes (Asteroidea) are rich in polar steroids with diverse structural characteristics. The structural modifications of star fish steroids occur at 3ß, 4ß, 5α, 6α (or ß), 7α (or ß), 8, 15α (or ß) and 16ß positions of the steroidal nucleus and in the side chain. Widely found polar steroids in starfishes include polyhydroxysteroids, steroidal sulfates, glycosides, steroid oligoglycosides etc. Bioactivity of these steroids is less studied; only a few reports like antibacterial, cytotoxic activity etc. are available. In continuation of our search for bioactive molecules from natural sources, we undertook in silico screening of steroids from star fishes against Bcl-2 and CDK-4/Cyclin D1 - two important targets of progression and proliferation of cancer cells. We have screened 182 natural steroids from star fishes occurring in different parts of the world and their 282 soft-derivatives by in silico methods. Their physico-chemical properties, drug-likeliness, binding potential with the selected targets, ADMET (absorption, distribution, metabolism, toxicity) were predicted. Further, the results were compared with those of existing steroidal and non steroidal drugs and inhibitors of Bcl-2 and CDK-4/Cyclin D1. The results are promising and unveil that some of these steroids can be potent leads for cancer treatments.

Effect of strontium in combination with fluoride on enamel remineralization in vitro.

Previous studies showed that strontium (Sr) as well as fluoride (F) can enhance enamel remineralization. The aim of this study was to evaluate the effects of Sr in combination with F on enamel remineralization in vitro. Sixty enamel specimens obtained from caries free human premolars were demineralised to produce caries-like lesions. Half of each lesion was covered with nail varnish as an untreated control. The specimens were then randomly divided into F and Sr+F treatment groups. The F group was exposed to remineralizing solutions (1.5mM CaCl(2), 0.9 mM KH(2)PO(4)) containing 1 ppm, 0.1 ppm or 0.05 ppm F. The Sr+F treatment group was exposed to the same solutions including 10 ppm Sr. After 2 weeks, lesion depth, mineral loss and percentage enamel remineralization were determined using transversal microradiography. There was a significant decrease in mineral loss in all groups (p<0.001). Lesion depth was significantly reduced for all groups (p<0.05) with the exception of group F. Remineralization was significantly affected by F concentration (p=0.000). The participation of Sr resulted in a significant enhancement of remineralization (p<0.001) with a synergistic effect of the Sr+F combination (p<0.01). It was concluded that while the remineralizing process was affected by the concentration of F, there was also an interaction between F and Sr when they were used in conjunction.

The effect of fluoridation and its discontinuation on fluoride profiles in the alveolar bone of rat.

We investigated the effect of fluoridation and its discontinuation on fluoride content in the alveolar portion of the mandible in rats. Drinking water with three different fluoride contents (0, 50, 100 ppmF) was given to rats for three different periods (4, 13 and 25 weeks). Fluoride concentrations were measured in the crest, the middle, and the apical parts of the alveolar bone and in the body of the mandible. Furthermore, after fluoridated drinking water was given to rats for 4 or 13 weeks, distilled water was given to them for 21 or 12 weeks respectively; and the effect of the discontinuation on fluoride profiles was investigated. Layer samples were analyzed by abrasive microsampling. Fluoride and phosphorus concentrations were determined by ion-specific electrode and colorimetric procedures, respectively. There was an increase in fluoride concentrations in the mandible in proportion to the fluoride content in the drinking water and the duration of fluoridation. After fluoridation was discontinued, fluoride concentrations in the surface layers of the mandible presented a decrease. Among the four different parts of the mandible, the upper part of the alveolar bone and the alveolar crest part presented the highest rates of reduction. The relative reduction rate of fluoride concentration was closely related to the duration of discontinuation. The alveolar crest was affected most by the discontinuation of fluoridation, presenting the greatest reduction.

Fluoride profiles in premolars after different durations of water fluoridation in Ho Chi Minh City, Vietnam.

In order to evaluate the uptake of fluoride (F) into tooth structures following water fluoridation in Ho Chi Minh City, F concentrations were measured from the enamel surface through the enamel-dentine junction (EDJ) to dentine close to the pulp in premolars. Calcification had been completed in all the teeth before fluoridation. Samples were obtained from fluoridated areas of the City ([F]: 0.7 parts/10(6)) after 3 years (eight samples), 6 years (eight samples) and 8 years (nine samples). Samples were also taken from regions outside the fluoridated water supply of the City in 1998 after 8 years of water fluoridation. An abrasive microsampling method was used to determine the profiles of fluoride and phosphorus concentrations. In enamel surfaces, F concentrations tended to increase with increasing periods of fluoridation. In dentine close to the pulp, F concentrations also increased statistically significantly with the increasing duration of fluoridation of water (between 6 and 3 years: P=0.006; between 8 and 3 years: P=0.001; between 8 years and 8 years without F: P=0.0001). It was concluded that F concentrations in enamel and dentine had tended to increase gradually with the duration of water fluoridation in Ho Chi Minh City. The increase was most obvious in dentine near the junction with the pulp.