Genetic polymorphisms associated with developmental defects of enamel: A systematic review.

BACKGROUND: Polymorphisms in genes related to enamel formation and mineralization may increase the risk of developmental defects of enamel (DDE). AIM: To evaluate the existing literature on genetic polymorphisms associated with DDE. DESIGN: This systematic review was registered in the PROSPERO (CRD42018115270). The literature search was performed in PubMed, Scopus, Web of Science, LILACS, BBO, Cochrane Library, and in the gray literature. Observational studies assessing the association between DDE and genetic polymorphism were included. The Newcastle-Ottawa Scale was used to assess the risk of bias. RESULTS: One thousand one hundred and forty-six articles were identified, and 28 met the inclusion criteria. Five studies presented a low risk of bias. Ninety-two genes related to enamel development, craniofacial patterning morphogenesis, immune response, and hormone transcription/reception were included. Molar-incisor hypomineralization (MIH) and/or hypomineralization of primary second molars (HPSM) were associated with 80 polymorphisms of genes responsible for enamel development, immune response, morphogenesis, and xenobiotic detoxication. A significant association was found between the different clinical manifestations of dental fluorosis (DF) with nine polymorphisms of genes responsible for enamel development, craniofacial development, hormonal transcription/reception, and oxidative stress. Hypoplasia was associated with polymorphisms located in intronic regions. CONCLUSION: MIH, HPSM, DF, and hypoplasia reported as having a complex etiology are significantly associated with genetic polymorphisms of several genes.

Epigallocatechin-3-gallate attenuates fluoride induced apoptosis via PI3K/FoxO1 pathway in ameloblast-like cells.

Fluoride is a double-edged sword. It was widely used for early caries prevention while excessive intake caused a toxicology effect, affected enamel development, and resulted in dental fluorosis. The study aimed to evaluate the protective effect and mechanism of Epigallocatechin-3-gallate (EGCG) on the apoptosis induced by fluoride in ameloblast-like cells. We observed that NaF triggered apoptotic alterations in cell morphology, excessive NaF arrested cell cycle at the G1, and induced apoptosis by up-regulating Bax and down-regulating Bcl-2. NaF activated the insulin-like growth factor receptor (IGFR), and phosphatidylinositol-3-hydroxylase (p-PI3K), while dose-dependently down-regulating the expression of Forkhead box O1 (FoxO1). EGCG supplements reversed the changes in LS8 morphology, the cell cycle, and apoptosis induced by fluoride. These results indicated that EGCG possesses a protective effect against fluoride toxicity. Furthermore, EGCG suppressed the activation of p-PI3K and the down-regulation of FoxO1 caused by fluoride. Collectively, our findings suggested that EGCG attenuated fluoride-induced apoptosis by inhibiting the PI3K/FoxO1 signaling pathway. EGCG may serve as a new alternative method for dental fluorosis prevention, control, and treatment.

Comparison of case-based and lecture-based learning in dental fluorosis diagnostic ability with visual analog scale assessment.

OBJECTIVE: This study aimed to compare the impact of case-based learning (CBL) versus lecture-based learning (LBL) on dental students' clinical decision-making regarding DF severity using Visual Analog Scale (VAS) scoring. METHODS: Eighty first-year graduate dental students were randomly assigned to either the CBL (n = 38) or LBL (n = 42) groups. Both groups received instruction on DF diagnosis, with CBL involving small group sessions analyzing real cases and LBL involving traditional lectures. Effectiveness was assessed by presenting 32 dental fluorosis cases with Thylstrup-Fejerskov Index (TSIF) scores ranging from 0 to 7 through slide presentations to both groups for VAS assessment. Five evaluators of each group randomly selected were asked to repeat the rating 2 weeks later. Statistical analysis included two-way ANOVA for group and gender differences, intra-class correlation coefficient (ICC) for reliability, and Spearman correlation coefficients for validity. RESULTS: Variations in VAS scores were observed between CBL and LBL groups, with no significant gender impact. Excellent inter- and intra-evaluator agreement was found for VAS scoring in both groups, indicating its reliability. Validation against established indices (such as DI and TSIF) demonstrated strong correlations, with CBL students exhibiting higher correlations. CONCLUSIONS: CBL enhances students' clinical decision-making and proficiency in DF diagnosis, as evidenced by more consistent and accurate VAS scoring compared to LBL. These findings highlight the importance of innovative educational strategies in dental curricula, with implications for improving training quality and clinical outcomes. TRIAL REGISTRATION: The study was registered at the Clinical Research Center, Hospital of Stomatology, Wuhan University (Registration code: HGGC-036).

A review of oral health promotion programmes in Eastern Mediterranean Region.

Background: Oral health conditions, such as dental caries, periodontal disease, tooth loss, dental fluorosis, dental trauma, and oral cancer, are prevalent in the WHO Eastern Mediterranean Region. However, there has been no systematic review of oral health promotion interventions in the region. Aims: To review existing literature on oral health promotion programmes in the Eastern Mediterranean Region and recommend improvements for the future. Method: We reviewed on PubMed and Google Scholar 61 articles published in the Eastern Mediterranean Region between 2010 and 2023. Quality assessment of included studies was performed using established criteria. We used the content analysis approach to create appropriate themes from the studies and to document meaningful conclusions about oral health promotion. Results: Majority of the studies were cross-sectional, a few were randomized controlled, quasi-experimental, longitudinal studies, or reviews. Oral health problems identified included poor oral health knowledge, dental caries, periodontal disease, tooth loss, dental fluorosis, and oral cancer. Although oral disorders were common in most of the countries, very few have implemented oral health promotion programmes. Conclusion: We recommend prioritization of oral health promotion programmes in the Eastern Mediterranean Region to tackle the diverse oral health challenges. To be effective, such programmes should be region- and context-specific. More studies on oral health promotion are needed in the region.

Association between fluoride intake from drinking water and severity of dental fluorosis in Northern and Western Mexico: systematic review and meta-analysis.

BACKGROUND: Dental fluorosis (DF) is caused by excessive exposure to fluoride during odontogenesis and leads to various changes in the development of tooth enamel. Some regions in Mexico are considered endemic fluorosis zones due to the high fluoride content in drinking water. The objective of this study was to perform a systematic review and meta-analysis to identify the association between the concentration of fluoride in drinking water and the severity of dental fluorosis in northern and western Mexico. METHODS: This protocol was registered in the PROSPERO database (ID: CRD42023401519). The search for information was carried out in the PubMed/Medline, Scopus, SpringerLink, and Google Scholar databases between January 2015 and October 2023. The overall relative risk was calculated using the inverse of variance approach with the random effects method. The RoB 2.0 tool was used to construct risk plots. RESULTS: Eleven articles were analyzed qualitatively, and most of the included studies presented at least one level of DF severity; six articles were analyzed quantitatively, dividing them into two regions. In North region it was observed a higher prevalence of severe TF cases, corresponding to ≥ TF 5 category (4.78) [3.55, 6.42]. In the West region, most of the included studies presented a higher prevalence of less severe cases, corresponding to ≤ TF 4, in comparison with the North region (0.01) [0.00, 0.52], interpreted as a protective effect. CONCLUSION: The concentrations of fluorides in drinking water are reportedly high in these regions and are directly related to the severity of dental fluorosis experienced by the inhabitants. In the Northern region exists a major concentration of fluoride in drinking water compared with the Western region as well as a prevalence of higher severity cases of dental fluorosis.

Tooth whitening: current status and prospects.

As a safe, effective, economical, and convenient technique, tooth whitening is one of the most popular treatments for improving tooth discoloration. This review summarizes the theoretical and recent research developments in the classification and mechanisms of tooth discoloration, as well as the principles, agents, effects, and side effects of tooth whitening techniques. The aim is to provide a basis for the clinical treatment of tooth whitening techniques and to suggest possible new ideas for further research. The accepted mechanism of whitening is the redox reaction of oxides in the whitening reagent, and the whitening effect is remarkable. However, side effects such as tooth sensitivity and irritation of gum and other oral soft tissues can still occur. It is recommended that more monitoring be carried out in the clinic to monitor these side effects, and care should be taken to protect the soft tissues in the mouth during office whitening procedures. Furthermore, there is a need to develop new additives or natural whitening products to reduce the occurrence of side effects.

Is Italian Dentists' Knowledge of Enamel Development Defects Adequate? A Nationwide Survey.

OBJECTIVES: Correct identification and management of Developmental Defects of Enamel (DDEs) are essential to provide the best possible treatment. The present survey aims to investigate Italian dentists' knowledge of DDEs, their ability to recognise the different clinical pictures, and to choose the most appropriate clinical approach. METHODS: A cross-sectional survey was planned based on a questionnaire including 27 closed-ended questions, and that proposed 4 clinical pictures, molar incisor hypomineralisation (MIH), amelogenesis imperfecta (AI), dental fluorosis (DF), and an initial caries lesion (ICL). It was distributed by e-mail to all Italian dentists (N = 63,883) through the Italian Federation of Doctors and Dentists. Discrete variables were expressed as absolute and relative frequencies (%). A multivariate analysis assessed whether socio-demographic variables correlated with the answers' truthfulness. RESULTS: About 5017 questionnaires were included and analysed. Although 90.19% of the sample stated that they had received information on DDEs, a significant percentage did not recognise MIH (36.36%), AI (48.34%), DF (71.50%), and ICL (46.62%). Only 57.07% correctly classified enamel hypomineralisation as a qualitative defect, and even fewer, 54.45%, classified enamel hypoplasia as a quantitative defect. According to the logistic regressions, female dentists, dentists who treat mainly children and received information about DDEs, were more likely to recognise the 4 clinical pictures (P < .01). CONCLUSIONS: Italian dentists showed many knowledge gaps on DDEs that need to be filled; those who received formal training were more capable of correctly identifying the defects and were more likely to prescribe an appropriate management approach for the defects. CLINICAL SIGNIFICANCE: Increasing university courses and continuing education on diagnosing and managing DDEs seems reasonable to fill the knowledge gap on DDEs.

Effect of Calcium Supplementation and TMEM16A Inhibition on Endoplasmic Reticulum Stress Induced by Dental Fluorosis in Mice.

BACKGROUND: Dental fluorosis is a discoloration of the teeth caused by the excessive consumption of fluoride. It represents a distinct manifestation of chronic fluorosis in dental tissues, exerting adverse effects on the human body, particularly on teeth. The transmembrane protein 16a (TMEM16A) is expressed at the junction of the endoplasmic reticulum and the plasma membrane. Alterations in its channel activity can disrupt endoplasmic reticulum calcium homeostasis and intracellular calcium ion concentration, thereby inducing endoplasmic reticulum stress (ERS). This study aims to investigate the influence of calcium supplements and TMEM16A on ERS in dental fluorosis. METHODS: C57BL/6 mice exhibiting dental fluorosis were subjected to an eight-week treatment with varying calcium concentrations: low (0.071%), medium (0.79%), and high (6.61%). Various assays, including Hematoxylin and Eosin (HE) staining, immunohistochemistry, real-time fluorescence quantitative polymerase chain reaction (qPCR), and Western blot, were employed to assess the impact of calcium supplements on fluoride content, ameloblast morphology, TMEM16A expression, and endoplasmic reticulum stress-related proteins (calreticulin (CRT), glucose-regulated protein 78 (GRP78), inositol requiring kinase 1α (IRE1α), PKR-like ER kinase (PERK), activating transcription factor 6 (ATF6)) in the incisors of mice affected by dental fluorosis. Furthermore, mice with dental fluorosis were treated with the TMEM16A inhibitor T16Ainh-A01 along with a medium-dose calcium to investigate the influence of TMEM16A on fluoride content, ameloblast morphology, and endoplasmic reticulum stress-related proteins in the context of mouse incisor fluorosis. RESULTS: In comparison to the model mice, the fluoride content in incisors significantly decreased following calcium supplements (p < 0.01). Moreover, the expression of TMEM16A, CRT, GRP78, IRE1α, PERK, and ATF6 were also exhibited a substantial reduction (p < 0.01), with the most pronounced effect observed in the medium-dose calcium group. Additionally, the fluoride content (p < 0.05) and the expression of CRT, GRP78, IRE1α, PERK, and ATF6 (p < 0.01) were further diminished following concurrent treatment with the TMEM16A inhibitor T16Ainh-A01 and a medium dose of calcium. CONCLUSIONS: The supplementation of calcium or the inhibition of TMEM16A expression appears to mitigate the detrimental effects of fluorosis by suppressing endoplasmic reticulum stress. These findings hold implications for identifying potential therapeutic targets in addressing dental fluorosis.

Sociodemographic, biological, and timing characteristics of dental caries and fluorosis using mixed-type cluster analysis on 12-year-olds in Ho Chi Minh city, 1989-2019.

Objectives: In 1990, Ho Chi Minh City started Community water fluoridation (CWF) at 0.7 ppm F, and in 2000, it was adjusted to 0.5 ppm F. Here, we analyzed dental caries and fluorosis data in Ho Chi Minh City to explore commonalities associated with CWF among 12-year-old children. Methods: Dental caries and fluorosis data were collected in 1989, 2003, 2012, and 2019 (N = 4773). Trained dentists scored dental caries using the WHO detection criteria and fluorosis using Dean's Fluorosis Index. We used these data and the k-prototypes method by the R package to identify clusters of participants with shared clinical and water fluoride levels. Results: We used datasets 1 (4773 participants) and 2 (4194 participants, missing fluorosis data in 1989). K-prototypes analysis identified three clusters in each dataset. Cluster 1, with 80 % of the sample at 0.5 ppm F area characterized by low caries and fluorosis scores. Cluster 2 with 60 % of the sample non-fluoridated area had high caries and low fluorosis scores. Cluster 3, with 75 % of the sample in 0.7 ppm area, had low caries but borderline high fluorosis scores. Conclusion: Identifying three clusters based on clinical and environmental scores supports the decision to fluoridate the water to prevent caries (0-0.7 ppm) and the shift from 0.7 to 0.5 ppm to keep the caries preventive effect while reducing the risk of fluorosis. Clinical significance: Our results support the effectiveness of CWF in preventing dental caries and the appropriateness of changing the F concentration to reduce the risk of fluorosis while maintaining its effectiveness.

Systematic review of epidemiological and toxicological evidence on health effects of fluoride in drinking water.

INTRODUCTION: Fluoride is a naturally occurring substance that is also added to drinking water, dental hygiene products, and food supplements for preventing dental caries. Concerns have been raised about several other potential health risks of fluoride. OBJECTIVE: To conduct a robust synthesis of evidence regarding human health risks due to exposure to fluoride in drinking water, and to develop a point of departure (POD) for setting a health-based value (HBV) for fluoride in drinking water. METHODS: A systematic review of evidence published since recent reviews of human, animal, and in vitro data was carried out. Bradford Hill considerations were used to weigh the evidence for causality. Several key studies were considered for deriving PODs. RESULTS: The current review identified 89 human studies, 199 animal studies, and 10 major in vitro reviews. The weight of evidence on 39 health endpoints was presented. In addition to dental fluorosis, evidence was considered strong for reduction in IQ scores in children, moderate for thyroid dysfunction, weak for kidney dysfunction, and limited for sex hormone disruptions. CONCLUSION: The current review identified moderate dental fluorosis and reduction in IQ scores in children as the most relevant endpoints for establishing an HBV for fluoride in drinking water. PODs were derived for these two endpoints, although there is still some uncertainty in the causal weight of evidence for causality for reducing IQ scores in children and considerable uncertainty in the derivation of its POD. Given our evaluation of the overall weight of evidence, moderate dental fluorosis is suggested as the key endpoint until more evidence is accumulated on possible reduction of IQ scores effects. A POD of 1.56 mg fluoride/L for moderate dental fluorosis may be preferred as a starting point for setting an HBV for fluoride in drinking water to protect against moderate and severe dental fluorosis. Although outside the scope of the current review, precautionary concerns for potential neurodevelopmental cognitive effects may warrant special consideration in the derivation of the HBV for fluoride in drinking water.

Identifying the oral microbiome of adolescents with and without dental fluorosis based on full-length 16S rRNA gene sequencing.

Dental fluorosis, resulting from long-term environmental exposure to fluoride, is prevalent among diverse populations worldwide. Severe fluorosis not only compromises the aesthetic appeal of teeth but also impairs their functionality. This study aims to investigate the oral microbiome in dental fluorosis and the health individuals of adolescents living in the endemic fluorosis area of Guizhou, China through full-length 16S rDNA sequencing. Fourty-six individuals meet the sampling criteria, and we divided these samples into the following groups: a healthy group (H = 23) and a dental fluorosis group (F = 23), and two subgroups of Miao ethnicity: a healthy Miao group (Hm = 13) and a dental fluorosis Miao group (Fm = 15). A total of 660,389 high-quality sequences were obtained, and 12,007 Amplicon Sequence Variants (ASVs) were identified, revealing significant variations in oral microbiome between Fm and Hm groups. The composition of oral microbiota was similar between the H and F groups. At the genus level, Pseudopropionibacterium and at the species level, Streptococcus oralis_subsp.dentisani_clade_058 were less abundant in group F than in group H (P < 0.05). Further analysis revealed that the abundance of Capnocytophaga gingivalis and Kingella denitrificans was significantly lower in Fm fluorosis patients than in the Hm group (P < 0.05). Based on the LEfSe analysis, the potential core biomarkers in the oral of Fm fluorosis patients were identified at different taxonomic levels, ranging from phylum to species. These include Gammaproteobacteria, Prevotella sp_HMT_304, Gemella sanguinis, and Gracilibacteria_(GN02). Network analysis revealed that the microbiota in the fluorosis group exhibited more complex interactions with each other than the healthy group. Notably, within the Hm group, the potential biomarkers Capnocytophaga gingivalis and Kingella denitrificans exhibited a positive correlation. Finally, we employed PICRUSt2 analysis to explore the abundance clustering of the top 30 functional units in each sample, and we found that the metabolic pathway compositions of the four groups were similar. In summary, our findings suggest that the microbial composition of plaque in Hm patients with dental fluorosis is significantly altered, and we identified the potential marker microorganisms that contribute to these changes.

Spatial-Temporal Analysis of Drinking Water Type of Endemic Fluorosis - China, 2009-2022.

What is already known about this topic?: Endemic fluorosis, caused by high fluoride levels in drinking water, has been a significant health issue in rural areas of China for many decades. What is added by this report?: There has been a notable decline in the detection rate of dental fluorosis in children aged 8-12 years in drinking water fluorosis areas across the country from 2009 to 2022. While 14 provincial-level administrative divisions are classified as low-probability clusters, Tianjin remains classified as a high-probability cluster. What are the implications for public health practice?: The current policy for preventing and controlling endemic fluorosis in China needs adjustment. Rather than focusing solely on regions with high incidence, there should be a shift towards monitoring and early warning of fluoride exposure. Additionally, local containment measures should be intensified.

Potential risk of dental fluorosis associated with different baby formulas and water brands marketed in Spain.

Despite efforts to promote breastfeeding, many babies aged <6 months receive only baby formula reconstituted with bottled water. The intake of high levels of fluoride during amelogenesis has been associated with hypomineralization of the tooth enamel, with aesthetic and mechanical repercussions. The objective of this study was to determine the potential risk of dental fluorosis associated with baby formulas marketed in Spain. We measured 26 baby formulas frequently consumed in Spain; 17 brands for babies aged <6 months, 5 for those aged >6 months, and 4 ready-to-use brands. They were prepared with 4 types of water: distilled water and three brands of bottled water with different levels of fluoride. The fluoride concentration (mg/L or ppm F) was measured with an ion-specific electrode coupled to an ion analyzer. Each sample was prepared according to the manufacturer's instructions and analyzed in triplicate. A descriptive analysis was carried out. The minimum fluoride level found was 0.04 mg/L and the maximum was 1.02 mg/L. Considering the daily intake of these formulas, none exceeded the clinically-acceptable daily dose limit risk for fluorosis (0.07 mg F/day/kg body weight) when mixed with bottled water with a low level of fluoride (0.1 mg/L). However, when the same brands of formula were reconstituted with bottled water with a higher fluoride content (0.99 mg/L), they all exceeded the daily dose limit for the risk of fluorosis. As the potential risk of dental fluorosis associated with the formulas tested depends exclusively on the fluoride concentration of the waters used for reconstitution, formula packaging should contain a warning.

ERS Mediated by GRP-78/PERK/CHOP Signaling Is Involved in Fluoride-Induced Ameloblast Apoptosis.

Fluoride can be widely ingested from the environment, and its excessive intake could result in adverse effects. Dental fluorosis is an early sign of fluoride toxicity which can cause esthetic and functional problems. Though apoptosis in ameloblasts is one of the potential mechanisms, the specific signal cascade is in-conclusive. High-throughput sequencing and molecular biological techniques were used in this study to explore the underlying pathogenesis of dental fluorosis, for its prevention and treatment. A fluorosis cell model was established. Viability and apoptosis rate of mouse ameloblast-derived cell line (LS8 cells) was measured using cell counting kit-8 (CCK-8) assay and flow cytometry analysis. Cells were harvested with or without 2-mM sodium fluoride (NaF) stimulation for high-throughput sequencing. Based on the sequencing data, subcellular structures, endoplasmic reticulum stress (ERS), and apoptosis related biomarkers were verified using transmission electron microscopy, quantitative real-time polymerase chain reaction, and Western blotting techniques. Expression of ERS markers, apoptosis related proteins, and enamel formation enzymes were detected using Western blotting after addition of 4-phenylbutyrate (4-PBA). NaF-inhibited LS8 cells displayed time- and dose- dependent viability. Additionally, apoptosis and morphological changes were observed. RNA-sequencing data showed that protein processing in endoplasmic reticulum was obviously affected. ERS and apoptosis were induced by excessive NaF. Downregulation of kallikrein-related peptidase 4 (KLK4) was also observed. Inhibition of ERS by 4-PBA rescued the apoptotic and functional protein changes in cells. Excessive fluoride induces apoptosis by activating ERS, which is mediated by GRP-78/PERK/CHOP signaling. Key proteinase is present in maturation-stage enamel; KLK4 was also affected by fluoride, but rescued by 4-PBA. This study presents a possibility for therapeutic strategies for dental fluorosis, while further exploration is required.

Health risk assessment in an area of dental fluorosis disease from high fluoride drinking water: a case study from southeastern Türkiye.

This study focuses on identifying fluoride (F‒) concentrations and its health risk assessment (HRA) in drinking water sources in south-eastern Türkiye. Groundwater quality was assessed using some graphical approaches such as Schoeller and Piper diagrams and GIS mapping. Average daily exposure dosages through oral and dermal contact exposure routes were considered to determine the potential health risk of F‒ in groundwater. Groundwater samples were taken from 53 points in spring, summer, autumn, and winter seasons. The results showed that the average annual F‒ concentrations in water resources in the study area were 0.26‒3.62 mg/L. According to the HRA results, the highest F‒ health risk in this region was observed in children, followed by teenagers and adults. This study indicated that there is a strong relationship between the high health risk (4.28 > 3.5) in children and dental fluorosis caused by high F‒ concentration in groundwater.

TGF-ß1/Smad3 Signaling Is Required to Alleviate Fluoride-Induced Enamel Hypomineralization.

Excessive fluoride intake during enamel development can affect enamel mineralization, leading to dental fluorosis. However, its potential mechanisms remain largely unexplored. In the present study, we aimed to investigate the impact of fluoride on the expressions of RUNX2 and ALPL during mineralization and the effect of TGF-ß1 administration on fluoride treatment. A dental fluorosis model of newborn mice and an ameloblast cell line ALC were both used in the present study. The mice of the NaF group, including the mothers and newborns, were fed with water containing 150 ppm NaF after delivery to induce dental fluorosis. The mandibular incisors and molars showed significant abrasion in the NaF group. Immunostaining, qRT-PCR, and Western blotting analysis indicated that exposure to fluoride markedly down-regulated RUNX2 and ALPL in mouse ameloblasts and ALCs. Besides, fluoride treatment significantly decreased the mineralization level detected by ALP staining. Furthermore, exogenous TGF-ß1 up-regulated RUNX2 and ALPL and promoted mineralization, while the addition of SIS3 could block such TGF-ß1-induced up-regulation. In TGF-ß1 conditional knockout mice, the immunostaining of RUNX2 and ALPL was weaker compared with wild-type mice. Exposure to fluoride inhibited the expressions of TGF-ß1 and Smad3. Co-treatment of TGF-ß1 and fluoride up-regulated RUNX2 and ALPL compared with the fluoride alone treatment, promoting mineralization. Collectively, our data indicated that TGF-ß1/Smad3 signaling pathway was necessary for the regulatory effects of fluoride on RUNX2 and ALPL, and the fluoride-induced suppression of ameloblast mineralization was mitigated by activating TGF-ß1/Smad3 signaling pathway.

Effect of Long Non-coding RNA and DNA Methylation on Gene Expression in Dental Fluorosis.

In the process of tooth development, the interaction between genetic information, epigenetic inheritance, and environment jointly affects the teeth formation. At present, the mechanism of dental fluorosis is rarely studied from transcriptomics, and there is no report on epigenetic perspective. In the study, SD rats were randomly divided into dental fluorosis group and control group fed with NaF (150 mg/L) or distilled water for 8 weeks. After 3.5 days of birth, the RNAs or DNA of rat mandibular molars were detected by RNA-seq or MethylTarget, respectively. The results demonstrated that a total of 1723 differentially expressed genes (DEGs) and 2511 differential expression lncRNAs (DE-lncRNAs) were mainly involved in the ion channels, calcium ion transport, and immunomodulatory signaling pathways. ATP2C1 and Nr1d1, which were related to Ca2+ transport, cellular calcium homeostasis, endoplasmic reticulum stress and immunity, may be the key genes in the formation of dental fluorosis. Notably, we also found that the immune response plays an important role in the formation of dental fluorosis, and a large amount of DEGs was enriched in immune regulation and NF-κB signaling pathways. Furthermore, the methylation levels of 13 sites were increased in Ago4, Atf3, Atp2c1, Dusp1, Habp4, and Mycl, while methylation levels of 5 CpG sites decreased in Ago4, Atp2c1, Habp4, and Traf6, and conformably, the expression of these genes have been significantly changed. This study comprehensively analyzed the occurrence mechanism of dental fluorosis from transcriptomics and epigenetics, so as to provide theoretical reference for further research.

Prevalence and risk factors of dental fluorosis among children aged 8-12 years in Shandong province of China.

This study was to investigate the prevalence and severity of children's dental fluorosis (DF) in Shandong and identified the potential risk factors for DF. A total of 87 villages in Shandong were investigated to calculate the prevalence of DF and Community Fluorosis Index (CFI) in 2018-2019. Six hundred and seventy children were enrolled to identify the potential risk factors using univariate and multivariate logistic regressions. Goodman-Kruskal Gamma was used to explore the factors related to the severity of DF. In 87 villages, 1249 of 8700 (14.36%) children still have DF. The prevalence of DF in most villages was below 40% in 2018-2019. Water fluorine concentration when selected for the study and urinary fluorine concentration were related to the risk of DF (P < 0.001). Some eating habits, like lower frequency of eating fresh vegetables, eggs, and beans, were associated with the risk of DF (P < 0.001). The high water fluorine concentration, and lower frequency of eating fresh vegetables, eggs, and beans were also related to the severity of DF (P < 0.001). DF in children in Shandong province is still a common endemic disease. This study tries to provide a useful guide for the prevention and control of DF.

Epidemiologia de la Fluorosis Dental en adolescentes de 12 años: Revisión sistemática

La prevalencia de fluorosis dental presenta una gran variabilidad a nivel mundial. Es necesario su análisis como parte de la vigilancia epidemiológica. Objetivo: Conocer la literatura disponible sobre prevalencia de fluorosis dental a la edad de 12 años en relación con el método de fluoruración comunitario utilizado. Metodología: Dos investigadoras realizaron una revisión sistemática de la literatura sin límites temporales siguiendo las pautas PRISMA, utilizando las bases de datos Pubmed, Cochrane, Scopus, BVS y Google Schoolar en idioma inglés, español, portugués e italiano. Resultados: Fueron incluidos 19 artículos de diseño transversal, 16 pertenecientes a comunidades que utilizan agua fluorurada, uno que utiliza sal fluorurada y 2 que comparan resultados entre comunidades que utilizan agua o sal fluorurada. Conclusiones: existe gran variabilidad en los reportes de prevalencia de fluorosis dental. Independientemente del método de fluoruración comunitario utilizado las lesiones de fluorosis de severidad leve son las más prevalentes.

Os relatos sobre a prevalência de fluorose dentária aos 12 anos apresentam grande variabilidade, não havendo unificação quanto ao uso dos índices. Independentemente do meio comunitário de fluoretação e do índice utilizado, a fluorose dentária leve é ​​a mais prevalente. Objetivo: Conhecer a literatura disponível sobre prevalência de fluorose dentária aos 12 anos em relação ao método comunitário de fluoretação utilizado. Metodologia: Dois pesquisadores realizaram uma revisão sistemática da literatura sem limites de tempo seguindo as diretrizes PRISMA, utilizando as bases de dados Pubmed, Cochrane, Scopus, BVS e Google Schoolar em inglês, espanhol, português e italiano. Resultados: Foram incluídos 19 artigos transversais, sendo 16 pertencentes a comunidades que utilizam água fluoretada, un sal fluoretada e 2 que comparam resultados entre comunidades que utilizam água fluoretada ou salgada. Conclusões: Há grande variabilidade nos relatos de prevalência de fluorose dentária. Independentemente do método de fluoretação comunitária utilizado, as lesões de fluorose de gravidade leve são as mais prevalentes.

The prevalence of dental fluorosis presents great variability worldwide. Its analysis is necessary as part of epidemiological surveillance Objective: To know the available literature on the prevalence of dental fluorosis among 12 years-old in relation to the community fluoridation method used. Methodology: Two researchers carried out a systematic review of the literature without time limits following the PRISMA guidelines, using the Pubmed, Cochrane, Scopus, BVS and Google Schoolar databases in English, Spanish, Portuguese and Italian. Results: 19 cross-sectional articles were included, 16 belonging to communities that use fluoridated water, one that use fluoridated salt and 2 that compare results between communities that use fluoridated water or salt. Conclusions: there is great variability in the reports of prevalence of dental fluorosis. Regardless of the community fluoridation method used, fluorosis lesions of mild severity are the most prevalent.

Relationship between dental fluorosis, dental caries and salivary levels of Streptococcus mutans.

Background and Objective: Worldwide, dental caries is an endemic infection and a significant public health problem. Fluoride reduces caries by helping to prevent demineralisation, by remineralising early carious lesions, and by decreasing the activity and growth of Streptococcus mutans (S. mutans). Fluoride intake above the optimal levels leads to a condition known as dental fluorosis, which causes hypo-mineralisation of the tooth. Some studies have revealed that the severity of fluorosis is directly proportional to caries, but some showed opposite results. Hence, with these contradicting results, this study was undertaken to identify the relationship between different grades of dental fluorosis, dental caries and the most common cariogenic bacteria S. mutans in saliva. Materials and Methods: A total of 90 subjects within 14- to 16-year age range were selected and categorised depending on the severity of fluorosis into three groups: group I (mild), group II (moderate) and group III (severe) based on modified Dean's fluorosis index criteria (1942). Unstimulated whole saliva samples were collected using the oral rinse technique and processed for quantification of S. mutans using Mitis Salivarius Bacitracin Agar medium. The number of colony-forming units (CFUs) was determined using a colony counter and expressed as ≥102, ≥103, ≥104 and ≥105 CFUs per ml of the sample, in accordance with the scale recommended by the manufacturer (HiMedia Laboratories). The severity of the caries was measured using decayed, missing or filled teeth (DMFT). Results: Observations revealed that the overall DMFT was directly proportional to the level of S. mutans titres. It was observed that 67 to 73% of participants showed fewer colony counts (102 and 103) with minimal DMFT scores and 27 to 33% showed higher counts (104 and 105) with higher DMFT scores in all the groups. 105 CFUs of group III showed the highest mean DMFT scores (2.9) with an increased number of involved subjects than group II (2.3) and group I (1.5). In all, male participants had higher bacterial titres and DMFT scores than females. Conclusion: Fluoride in the drinking water served as anticariogenic agent regardless of the severity of fluorosis. Severe fluorosis with a higher incidence of caries and increased CFUs of S. mutans clearly indicates the importance of preventive measures and early treatment to reduce the severity of fluorosis and prevalence of dental caries.