Multi-ancestry Genome-Wide Association Study of Early Childhood Caries.

Early childhood caries (ECC) is the most common non-communicable childhood disease. It is an important health problem with known environmental and social/behavioral influences that lacks evidence for specific associated genetic risk loci. To address this knowledge gap, we conducted a genome-wide association study of ECC in a multi-ancestry population of U.S. preschool-age children (n=6,103) participating in a community-based epidemiologic study of early childhood oral health. Calibrated examiners used ICDAS criteria to measure ECC with the primary trait using the dmfs index with decay classified as macroscopic enamel loss (ICDAS ≥3). We estimated heritability, concordance rates, and conducted genome-wide association analyses to estimate overall genetic effects; the effects stratified by sex, household water fluoride, and dietary sugar; and leveraged the combined gene/gene-environment effects using the 2-degree-of-freedom (2df) joint test. The common genetic variants explained 24% of the phenotypic variance (heritability) of the primary ECC trait and the concordance rate was higher with a higher degree of relatedness. We identified 21 novel non-overlapping genome-wide significant loci for ECC. Two loci, namely RP11-856F16 . 2 (rs74606067) and SLC41A3 (rs71327750) showed evidence of association with dental caries in external cohorts, namely the GLIDE consortium adult cohort (n=∼487,000) and the GLIDE pediatric cohort (n=19,000), respectively. The gene-based tests identified TAAR6 as a genome-wide significant gene. Implicated genes have relevant biological functions including roles in tooth development and taste. These novel associations expand the genomics knowledge base for this common childhood disease and underscore the importance of accounting for sex and pertinent environmental exposures in genetic investigations of oral health.

Detection of Caries Around Amalgam Restorations on Approximal Surfaces.

To evaluate the in vitro performance of the International Caries Detection and Assessment System (ICDAS) visual examination, bitewing radiography (BW), and the DIAGNOdent 2190, a pen-type laser fluorescence device (LFpen), in detecting caries around amalgam restorations on approximal surfaces. Approximal surfaces (N=136) of permanent posterior teeth (N=110) with Class II amalgam restorations were assessed twice by two experienced examiners using ICDAS, BW, and LFpen. The occurrence of proximal overhangs was also evaluated. The teeth were histologically prepared and assessed for caries extension. Different cutoff limits for the LFpen were used. Intraexaminer and interexaminer reproducibility showed moderate to good agreement for all the methods (weighted κ/intraclass correlation coefficient=0.40 to 0.87). The specificities at D1 (all visible lesions affecting enamel) and D3 (lesions extended into dentin) were, respectively, 0.41 and 0.82 for ICDAS, 0.70 and 0.82 for BW, and 0.77-0.89 and 0.88-0.94 for LFpen. The sensitivities were 0.80 and 0.52 for ICDAS, 0.56 and 0.51 for BW, and 0.04-0.23 and 0.01-0.02 for LFpen at D1 and D3, respectively. At the D1/D3 thresholds, the accuracy and the area under the receiver operating characteristic curve (Az) values were similar and statistically higher for ICDAS (0.65/0.68 and 0.633/0.688) and BW (0.64/0.68 and 0.655/0.719), respectively; whereas, LFpen presented lower accuracy (0.37-0.44/0.49-0.52) and Az (0.390-0.454/0.345-0.395) values. The occurrence of overhangs (26.8%) was shown to be irrelevant in determining the presence of secondary caries. The ICDAS and BW methods presented the best performance in detecting caries lesions affecting enamel and dentin on approximal surfaces of amalgam restorations.

Risk factors of caries progression in a Hispanic school-aged population.

The purpose of this Institutional Review Board-approved study was to identify risk factors of caries lesion progression in children enrolled in rural schools in Puerto Rico. A convenience sample of 408 children (5-13 yrs old) was examined at baseline and at 12 and 24 mos with the International Caries Detection and Assessment System (ICDAS). A total of 395 caregivers completed a 25-item questionnaire including socio-demographic, dietary, protective factors, disease experience, and access to care. Caries progression was significant (89% and 91% at 12 and 24 mos, respectively). Multiple-variable models for predicting children with lesion progression and numbers of lesions progressing were calculated for 2 outcome variables (any-progression vs. progression-toward-cavitation). Models developed had areas under the receiver operating characteristic (ROC) curve ranging between 0.70 and 0.79 and were very similar regardless of the outcome (progression criteria), prediction time (12-24 mos), or inclusion (or not) of previous caries experience. Significant predictors of disease progression collected through a parent-completed questionnaire included questions related to caries experience in the child or caregiver, and the caregiver's rating of the child's oral health.

An in vitro comparison between laser fluorescence and visual examination for detection of demineralization in occlusal pits and fissures.

It has been demonstrated that when excited by laser light carious enamel appears dark compared to luminescent sound enamel. The aim of this study was to compare the sensitivity and specificity of visual exams (V), laser fluorescence (LF) and dye-enhanced LF (DELF) for detecting demineralization in occlusal pits and fissures. The actual presence of lesions was determined by subsequent confocal laser microscopy (CM), which was compared to histology (H). Independent clinical examiners visually graded three sites on occlusal surfaces of extracted, human premolars as sound or carious and also rated the color of each graded site as: 0 = same as surrounding enamel; 1 = white; 2 = light brown, or 3 = brown/dark brown. An argon laser was used to illuminate the teeth for LF and DELF; the images were captured with a CCD camera and then analyzed. DELF images were captured after the teeth had been exposed to 0.075% sodium fluorescein. Sections were then cut from each specimen and analyzed by CM and H for the presence or absence of caries. Results showed that DELF (0.72) was significantly more sensitive (p<0.05) than LF (0.49) and V (0.03) for detecting caries, but there were no significant differences among the methods in specificity (V 1.00; LF 0.67; DELF 0.60). When color was used as an indication of caries in V (VC, sensitivity 0.47; specificity 0.70), V exams were not different from LF. The area under the ROC curve, using H as the gold standard and CM as the test, was 0.78. Results indicated that DELF was the best diagnostic tool and that VC and LF were equally effective as diagnostic methods, when color of fissures was included as an indication of demineralization in the visual exam.

Laser fluorescence detection of demineralization in artificial occlusal fissures.

Laser fluorescence (LF) has been used previously to detect early smooth-surface lesions. Although its use for detection of occlusal demineralization has been implicated, it has not been demonstrated. The aim of this study was to determine whether LF could detect demineralization in the base of artificial fissures. To employ LF for detection of occlusal demineralization an apparatus was devised to direct laser light into fissures and simultaneously detect fluorescence from the base of the fissures. Three groups (n = 40/group) of differing fissure types were prepared (straight wall, converging and diverging wall) with either a sound or lesioned base. One half of each group was examined with LF and dye-enhanced LF (DELF); the other half was examined with LF, exposed to plaque, examined with LF and DELF, air-polished and examined with DELF. All images were scored twice as either (1) carious; (2) sound, or (3) undetermined, by a group of 3 examiners. For fissures without plaque, the average sensitivity was higher for DELF (0.76) than for LF (0.54) (p < 0.05). Likewise, the average specificity was higher for DELF (0.64) than for LF (0.29) (p < 0.05). In the presence of plaque, sensitivity was higher for DELF (0.91) compared to LF (0.43); however, specificity was lower for DELF (0.05) compared to LF (0.55). When the fissures were air-polished and then examined with DELF, sensitivity averaged 0.82 (p < 0.05), and specificity increased consistently (average 0.51, p < 0.05). It was concluded that, in the absence of plaque, DELF was a better diagnostic tool than LF for detection of demineralization in artificial fissures.

Effect of two fluorescent dyes on color of restorative materials.

PURPOSE: The objective of these two studies was to determine whether the use of dyes in vitro would affect the shade of tooth-colored restorative materials. MATERIALS AND METHODS: Two separate studies were performed. In Study 1, four groups (N = 36) of restorative materials; (1) resin composite (RSN), (2) resin-modified glass ionomer (RMGI), (3) conventional glass ionomer (CGI), and (4) porcelain (PCN), were subjected to a rinse in either tap water (control) or Pyrromethene 556 dye at a concentration of 0.01M for 30 seconds or 0.0015 M for 60 seconds. They were then rinsed with 25% ethanol as would be done in clinical usage. In Study 2, the same restorative materials (except PCN) were subjected to a rinse in either tap water (control) (N = 8) or 0.0018 M sodium fluorescein (N = 8) for 60 seconds followed by a 5-second rinse with tap water. For both studies the specimens were analyzed for overall color changes (delta E*) comparing baseline to immediately post dye exposure and then again after 1, 4, 24, and 48-hour storage under running tap water. In addition, an image of each specimen, illuminated under an argon laser light (HGM), was captured with a miniature charged-coupled device (CCD) color camera at the same time intervals when the color was measured. These images were analyzed for fluorescence using computer assisted methods. A software program then computed the mean and standard deviation of the fluorescence values for each image. The data were analyzed using repeated measures ANOVA. RESULTS: delta E* values from baseline on specimens treated with either Pyrromethene 556 or sodium fluorescein were not significantly different from water at any time. The only exceptions were CGI specimens exposed to 0.01 M Pyrromethene 556 and RMGI exposed to sodium fluorescein which had a higher delta E* immediately after treatment, however this effect was reversed after 1-hour rinse. Mean delta E* was less than 2.69 for specimens treated with Pyrromethene 556 and less than 3.20 for specimens treated with sodium fluorescein after 1-hour rinse in water. When the mean fluorescence level, as determined by the computer, was averaged there was no difference between the Pyrromethene 556 treatments and control for RMGI, RSN and PCN after 4-hour rinse in water and, as time in storage increased, the fluorescence level decreased. CGI treated with 0.01 M Pyrromethene 556 was significantly more fluorescent even after 48-hour rinse in water. All specimens treated with sodium fluorescein were also significantly more fluorescent than control even after 48-hour rinse in water. It was concluded that exposure of these restorative materials to Pyrromethene 556 at 0.01 M or 0.0015 M or to 0.0018 M sodium fluorescein will not lead to any prolonged visually detectable color changes.