Mirror, mirror? An evaluation of identical twin mirroring in tooth crown morphology.

It has been estimated that 25% of monozygotic ("identical") twin pairs exhibit reverse asymmetry (RA) or "mirroring" of minor anatomical features as a result of delayed zygote division. Here, we examine whether identical twin mirroring accounts for patterns of dental asymmetry in a sample of monozygotic and dizygotic ("fraternal") twins. We focus on crown morphology to approach the following question: is there an association between dental RA frequency and twin type suggestive of the presence of mirror image twins in our sample? Data were collected from 208 deciduous and 196 permanent dentitions of participants of the University of Adelaide Twin Study using Arizona State University Dental Anthropology System standards. RA frequencies were compared across morphological complexes (deciduous, permanent), twin types (monozygotic, dizygotic), and traits. Fisher's exact tests were performed to formally evaluate the association between twin type and dental RA. Across the entire dataset, RA rates failed to exceed 8% for any twin type. In monozygotic twins, deciduous mirroring totaled 5.3% of observed cases, while permanent mirroring totaled 7.8% of observed cases. We found no statistically significant association between RA and twin type for any morphological character (p-value range: 0.07-1.00). Our results suggest the timing of monozygotic twin division does not explain the structure of asymmetry for our morphology dataset and that published estimates of identical twin mirroring rates may be inflated or contingent upon phenotype. Instead, rates reported for this sample more closely align with the proposed etiology of this condition.

Genetic and Morphological Variation in Hypodontia of Maxillary Lateral Incisors.

(1) Background: Hypodontia has a multifactorial aetiology, in which genetic factors are a major component. Associated with this congenital absence, the formed teeth may show differences in size and shape, which may vary with the specific genetic variants and with the location of the missing teeth. The aims of the present study were to investigate a specific variant of MSX1, derive morphometric tooth measurements in a sample of patients with isolated maxillary lateral incisor agenesis and matched controls, and model the findings. (2) Methods: Genotyping of the MSX1 rs8670 genetic variant and morphometric measurements with a 2D image analysis method were performed for 26 hypodontia patients and 26 matched controls. (3) Results: The risk of upper lateral incisor agenesis was 6.9 times higher when the T allele was present. The morphometric parameters showed significant differences between hypodontia patients and controls and between the unilateral and bilateral agenesis cases. The most affected crown dimension in the hypodontia patients was the bucco-lingual dimension. In crown shape there was significant variation the Carabelli trait in upper first molars. (4) Conclusions: The MSX1 rs8670 variant was associated with variations in morphological outcomes. The new findings for compensatory interactions between the maxillary incisors indicate that epigenetic and environmental factors interact with this genetic variant. A single-level directional complex interactive network model incorporates the variations seen in this study.

Genetic Correlation, Pleiotropy, and Molar Morphology in a Longitudinal Sample of Australian Twins and Families.

This study aims to expand our understanding of the genetic architecture of crown morphology in the human diphyodont dentition. Here, we present bivariate genetic correlation estimates for deciduous and permanent molar traits and evaluate the patterns of pleiotropy within (e.g., m1-m2) and between (e.g., m2-M1) dentitions. Morphology was observed and scored from dental models representing participants of an Australian twin and family study (deciduous n = 290, permanent n = 339). Data collection followed Arizona State University Dental Anthropology System standards. Genetic correlation estimates were generated using maximum likelihood variance components analysis in SOLAR v.8.1.1. Approximately 23% of deciduous variance components models and 30% of permanent variance components models yielded significant genetic correlation estimates. By comparison, over half (56%) of deciduous-permanent homologues (e.g., m2 hypocone-M1 hypocone) were significantly genetically correlated. It is generally assumed that the deciduous and permanent molars represent members of a meristic molar field emerging from the primary dental lamina. However, stronger genetic integration among m2-M1/M2 homologues than among paired deciduous traits suggests the m2 represents the anterior-most member of a "true" molar field. The results indicate genetic factors act at distinct points throughout development to generate homologous molar form, starting with the m2, which is later replaced by a permanent premolariform crown.

Patterns of heritability across the human diphyodont dental complex: Crown morphology of Australian twins and families.

OBJECTIVES: This study generates a series of narrow-sense heritability estimates for crown morphology of the deciduous and permanent dentition with two overarching aims. The first is to test the hypothesis that deciduous teeth provide a more faithful reflection of genetic information than their permanent successors. The second is to use quantitative genetic methods to evaluate assumptions underlying common data collection and analysis practices in biodistance research. MATERIALS AND METHODS: Dental morphology data were collected from longitudinal dental casts representing Australian twins and families using Arizona State Dental Anthropology System standards. Polygenic models and estimates of narrow-sense heritability were generated using SOLAR v.8.1.1. Each model considered age, sex, and age/sex interaction as covariates. RESULTS: Heritability estimates significantly differed from zero for the majority of morphological crown characters. Most estimates fell within the 0.4-0.8 range typically observed for crown morphology. Mean heritability was stable across the dental complex, but for paired homologues, permanent traits often yielded higher estimates than their deciduous counterparts. Results indicate directional asymmetry in environmental influence for crown morphology and inform biodistance "best practices" related to data collection and treatment. CONCLUSIONS: Overall, results for this sample support the use of crown morphology as a proxy for genetic variation in evolutionary research. This includes the deciduous dentition, which justifies the expansion of efforts to incorporate subadults into reconstructions of past microevolutionary processes. Results do not indicate that deciduous phenotypes more closely approximate underlying genotype, at least for deciduous/permanent homologues.

Compensatory interactions between developing maxillary anterior teeth in a sample of twins.

OBJECTIVES: The objective of this study was to revisit the topic of compensatory interactions between maxillary anterior teeth during development advanced by Sofaer et al. (1971). We addressed the hypotheses listed by Sofaer and colleagues using data derived from our Australian twin sample to investigate whether final tooth size in permanent maxillary central incisors and canines showed evidence of developmental compensation for adjacent missing or microdont lateral incisors. Such compensation is one factor interacting in the complex system of dental development. MATERIALS AND METHODS: A 2D image analysis system was used to measure crown height from the labial view, labiopalatal crown width from the incisal view, and mesiodistal crown width from both the labial (MDl) and incisal (MDi) views of the permanent maxillary central incisors and canines on the dental study model of twins enrolled in a longitudinal study of dental development. RESULTS: Developmental variations of maxillary lateral incisors influence the morphogenesis of the adjacent teeth. For example, individuals with one missing lateral incisor and one lateral incisor of average dimensions, had significantly larger central incisors than the control group for the MDl and MDi dimensions (p < 0.05). Of the 7 monozygotic twin pairs, 6 were discordant in maxillary anterior hypodontia and microdontia, and 13 out of 14 dizygotic twin pairs were discordant. CONCLUSIONS: This study provides further evidence of developmental interactions in the maxillary anterior region, partially supporting Sofaer and colleagues' hypotheses. These interactions are part of a complex adaptive system involving genetic, epigenetic and environmental factors.

Mild hypodontia is associated with smaller tooth dimensions and cusp numbers than in controls.

OBJECTIVE: The associations seen clinically between variations in tooth number, size and shape reflect the repetitive genetic interactions occurring between the epithelium and mesenchyme during the initiation and morphogenetic stages of the Complex Adaptive System that is dental development. The aim of this study was to investigate the clinical relationship further by comparing multiple crown parameters, including cusp numbers, between patients with mild hypodontia and controls in a Romanian sample. DESIGN: Digital images of dental casts of the permanent dentition from 28 patients with mild hypodontia and 28 controls were used. Measurements from the vestibular and occlusal surfaces were performed using a 2D image analysis method and cusps, including the Carabelli trait, were counted. Two-way analysis of variance was performed. RESULTS: The dimensions of the mild hypodontia group had smaller values than the controls, with many measurements being significantly different (significance values varied from p=0.049 to p=0.001). The most affected regions were the upper and lower anterior region in both sexes. Mesio-distal, bucco-lingual and occlusal area and perimeter dimensions were affected. Females from the hypodontia group had significantly less tricuspidated lower premolars when compared with the control group. Carabelli cusps were present in the hypodontia group less frequently, the difference being highly significant (p=0.0002) in women. CONCLUSIONS: The hypodontia patients presented with reduced crown dimensions and shape compared with controls. This is the first published study to demonstrate smaller cusp numbers in patients with hypodontia than in controls. The findings are compatible with a model of dental development as a Complex Adaptive System incorporating associations between tooth number, size and shape.

Diagnosis of Helicobacter pylori by carbon-13 urea breath test using a portable mass spectrometer.

CONTEXT: In the non-invasive detection of markers of disease, mass spectrometry is able to detect small quantities of volatile markers in exhaled air. However, the problem of size, expense and immobility of conventional mass spectrometry equipment has restricted its use. Now, a smaller, less expensive, portable quadrupole mass spectrometer system has been developed. Helicobacter pylori has been implicated in the development of chronic gastritis, gastric and duodenal ulcers and gastric cancer. OBJECTIVES: To compare the results obtained from the presence of H. pylori by a carbon-13 urea test using a portable quadrupole mass spectrometer system with those from a fixed mass spectrometer in a hospital-based clinical trial. METHODS: Following ethical approval, 45 patients attending a gastroenterology clinic at the Royal Liverpool University Hospital exhaled a breath sample into a Tedlar gas sampling bag. They then drank an orange juice containing urea radiolabelled with carbon and 30 min later gave a second breath sample. The carbon-13 content of both samples was measured using both quadrupole mass spectrometer systems. If the post-drink level exceeded the pre-drink level by 3% or more, a positive diagnosis for the presence of H. pylori was made. RESULTS: The findings were compared to the results using conventional isotope ratio mass spectrometry using a laboratory-based magnetic sector instrument off-site. The results showed agreement in 39 of the 45 patients. CONCLUSIONS: This study suggests that a portable quadrupole mass spectrometer is a potential alternative to the conventional centralised testing equipment. Future development of the portable quadrupole mass spectrometer to reduce further its size and cost is indicated, together with further work to validate this new equipment and to enhance its use in mass spectrometry diagnosis of other medical conditions.

Three-dimensional profilometric assessment of early enamel erosion simulating gastric regurgitation.

OBJECTIVES: A priority research area in minimal intervention dentistry is the characterization of the early stages of dental erosion. The aim of this in vitro study was to assess the effect of short, repetitive erosive challenges to human enamel over 2 min at pH 1.5 and 3.0 under conditions simulating gastric regurgitation. METHODS: Enamel surfaces were subjected to erosive challenges at pH 1.5 (Group 1, n=10) and pH 3.0 (Group 2, n=9) for periods of 30s (stage 1), 60s (stage 2) and 120 s (stage 3). Quantitative changes were assessed longitudinally by measuring the 3D average surface roughness (Sa) values using 3D confocal microscopy. Qualitative micrographic assessment of surface changes was also conducted by using environmental scanning electron microscopy. RESULTS: Linear mixed model analysis showed significant effects of the pH values (p<0.001) and the stages (p<0.001) on the observed Sa values. Post hoc tests showed significant increases in the Sa values between baseline and other stages in both groups (p<0.01). The mean Sa values also increased significantly from stage 1 to stage 2 in Group 1 (p<0.05). Micrographic analysis displayed severely etched enamel rods in Group 1, but only subtle changes in Group 2. CONCLUSIONS: The complexity of the enamel surface is influenced by both acid concentration (pH value) and duration of acid exposure during early stages of erosion. Erosion occurring under conditions simulating GORD can be detected in its initial stages, opening up the possibilities of early diagnosis and management of this condition. CLINICAL SIGNIFICANCE: Erosive tooth wear occurs progressively and insidiously, often creating complex treatment challenges. This emphasizes the need for early diagnosis and management in accordance with minimal intervention philosophy. Our findings provide a foundation for further research that could lead to the development of highly-sensitive clinical diagnostic tools and preventive strategies.

A large X-chromosomal deletion is associated with microphthalmia with linear skin defects (MLS) and amelogenesis imperfecta (XAI).

A female patient is described with clinical symptoms of both microphthalmia with linear skin defects (MLS or MIDAS) and dental enamel defects, having an appearance compatible with X-linked amelogenesis imperfecta (XAI). Genomic DNA was purified from the patient's blood and semiquantitative multiplex PCR revealed a deletion encompassing the amelogenin gene (AMELX). Because MLS is also localized to Xp22, genomic DNA was subjected to array comparative genomic hybridization, and a large heterozygous deletion was identified. Histopathology of one primary and one permanent molar tooth showed abnormalities in the dental enamel layer, and a third tooth had unusually high microhardness measurements, possibly due to its ultrastructural anomalies as seen by scanning electron microscopy. This is the first report of a patient with both of these rare conditions, and the first description of the phenotype resulting from a deletion encompassing the entire AMELX gene. More than 50 additional genes were monosomic in this patient.

The in vitro and in vivo validation of a mobile non-contact camera-based digital imaging system for tooth colour measurement.

OBJECTIVE: To assess the reproducibility of a mobile non-contact camera-based digital imaging system (DIS) for measuring tooth colour under in vitro and in vivo conditions. METHODS: One in vitro and two in vivo studies were performed using a mobile non-contact camera-based digital imaging system. In vitro study: two operators used the DIS to image 10 dry tooth specimens in a randomised order on three occasions. In vivo study 1:25 subjects with two natural, normally aligned, upper central incisors had their teeth imaged using the DIS on four consecutive days by one operator to measure day-to-day variability. On one of the four test days, duplicate images were collected by three different operators to measure inter- and intra-operator variability. In vivo study 2:11 subjects with two natural, normally aligned, upper central incisors had their teeth imaged using the DIS twice daily over three days within the same week to assess day-to-day variability. Three operators collected images from subjects in a randomised order to measure inter- and intra-operator variability. RESULTS: Subject-to-subject variability was the largest source of variation within the data. Pairwise correlations and concordance coefficients were > 0.7 for each operator, demonstrating good precision and excellent operator agreement in each of the studies. Intraclass correlation coefficients (ICCs) for each operator indicate that day-to-day reliability was good to excellent, where all ICC's where > 0.75 for each operator. CONCLUSION: The mobile non-contact camera-based digital imaging system was shown to be a reproducible means of measuring tooth colour in both in vitro and in vivo experiments.

The new Enamel Defects Index: testing and expansion.

The Enamel Defects Index (EDI) was created based on three innovative principles: (i) a basic level of the three major categories of defects; (ii) more detailed subcategories of each major category; and (iii) each category scored independently as present [1] or absent [0], simplifying decision making. The aim of this investigation was to further test the index in a number of applications and to expand it to record defect subtype and treatment need. Testing was undertaken by operators with different levels of clinical experience. A computer-assisted learning (CAL) package was developed for operator training and calibration. The index was also used on clinical photographs and high-resolution digital images of exfoliated and extracted teeth. Scoring of photographs revealed substantial intra-operator agreement. Training using the CAL package resulted in significant improvement in index use. Intra-operator reproducibility was good to excellent, and interoperator reproducibility was good for buccal surfaces on digital images. Index expansion allowed information on defect subtype, location, and treatment need to be gathered readily. The EDI has high reproducibility and allows more rapid and accurate data collection from clinical and in vitro studies than the Fédération Dentaire Internationale Developmental Defects of Enamel index.

Comparison of methods for measurement of hypoplastic lesions.

Enamel hypoplasia is a quantitative defect of enamel thickness. Methods previously used for its measurement have limitations in clinical studies. The aim of this study was to investigate new methods of measurement using image analysis. Lesions on 8 teeth affected by enamel hypoplasia were quantified from study models and impression surfaces using an image-analysis system. The measurements made included lesion area and tooth surface area; from these the proportion of tooth surface area affected was calculated. For comparison, manual measurement was performed on impression surfaces and study models, using digital callipers. Images were also acquired of lesions on 12 exfoliated teeth, and the lesion area and total tooth area were calculated. For assessment of intra-operator reliability, the +/-repeatability coefficient was calculated. Measurement of the surface of lesions direct from the exfoliated teeth gave the best results overall, followed by direct image analysis of the silicone impression.

Hypoplastic enamel defects and environmental stress in a homogeneous Romano-British population.

This study investigated hypoplastic enamel defects in a well-defined sample of Romano-Britons from the 3rd to the 5th century AD and compared the findings with a modern British sample investigated by the same authors. All 178 excavated skulls with intact dentitions were examined for hypoplastic defects using the Federation Dentaire International (FDI) Developmental Defects of Enamel Index criteria. Histopathological and microradiographic sections were prepared of 5 teeth. Hypoplastic defects were found in the teeth of 37% of skulls, with 25% having 4 or more teeth affected. The teeth most frequently involved were canines. Of the defects, 75% were horizontal grooves, 12.7% were pitting, and 7.1% were areas of missing enamel. The location of defects was 82% buccal, 16.5% lingual, and 1.2% occlusal. The reproducibility of diagnosis was 84%. Microscopic and microradiographic investigations showed areas of hypomineralization of enamel and wide zones of interglobular dentine related to the hypoplastic grooves. There was higher frequency, different morphology, and greater severity of hypoplastic enamel defects compared with the modern British sample. The defects may be related to repeated environmental stresses between the ages of 2 and 6 yr. Identified environmental stresses in these Romano-Britons, including high lead ingestion, poor nutrition, and recurrent infections, may be important etiological factors for the enamel defects.

A randomized controlled trial of outreach placement's effect on dental students' clinical confidence.

This randomized controlled trial compared the effects of outreach placement with traditional, exclusively dental school-based clinical experience on students' confidence in providing treatment for patients presenting with common dental problems. Senior dental students (n=49) were allocated at random to existing dental school-based clinics or placement in primary care clinics to work supervised by local dentists. At baseline, students self-rated their global confidence on a five-point scale. These measures were repeated at follow-up, augmented by a transition judgment and a then-test of confidence (asking students to look back and re-rate their pre-placement confidence). Groups were comparable at baseline. Follow-up scores for global confidence were similar between groups. The outreach group scored higher in the transition judgment (mean 3.7, SD 0.9) than the control group (mean 3.1, SD 1.1, P=0.05). In the then-test, on reflection, the outreach group considered their baseline ratings of confidence were overoptimistic (mean then-test scores 3.2, SD 0.9 and baseline 3.7, SD 0.5) while the control group thought theirs were accurate (mean then-test scores 3.8, SD 0.7 and baseline 3.6, SD 0.8, P=0.01). The findings suggest dental outreach training in primary care settings is more effective than dental school training alone in improving students' confidence in tackling clinical situations. The measurement of change in confidence is complicated by shifts in students' internal scales arising from insights gained on outreach.

Fluctuating dental asymmetry of multiple crown variables measured by an image analysis system.

OBJECTIVE: Fluctuating dental asymmetry as an indication of the effect of environmental insults during tooth development requires further investigation. The aim of this study was to assess asymmetry in tooth crown dimension for the increased range of parameters possible with image analysis compared to previous manual measurements. METHODS: Study models of 25 male and 25 female white Caucasian subjects were measured from buccal and occlusal views to determine mesio-distal, bucco-lingual, occluso-gingival and tooth surface area dimensions using image analysis. Method errors were assessed by the repeatability coefficient RC and asymmetry assessed calculating intra-class correlation coefficients between pairs of antimere teeth. RESULTS: The method errors calculated using RC were small. There was a high degree of symmetry with correlation coefficients with a range of 0.67-0.96 for linear measurements and 0.80-0.97 for tooth surface areas. No significant differences were found between the genders or between the upper and lower jaws. Asymmetry tended to follow a pattern in morphogenic tooth fields with the mesial tooth showing greater symmetry than the more distal. There were differences in the degree of symmetry among the variables measured with the mesio-distal dimension showing the greatest symmetry. CONCLUSIONS: This new technique was reliable and readily applicable, providing also a greater range of measurements. While the asymmetry generally followed a similar pattern to earlier manual studies, there were variations in the degree of symmetry between different parameters.

The patterning of hypodontia in a group of young adults in Sheffield, UK.

OBJECTIVE: Identifying the patterning of congenitally absent teeth will contribute to understanding the aetiology of hypodontia. The aim of this study was to investigate patterns of hypodontia in a group of young adults in Sheffield, UK involving specific tooth types, gender differences and maxilla:mandible and left:right correlations. METHODS: The patients were 198 unrelated Caucasian young adults (86 males and 112 females) with non-syndromic hypodontia. Statistical methods included the multivariate techniques of cluster analysis and principal components analysis and associated display devices of circular dotplots and biplots. Autologistic regression was used to model the relationship of the presence or absence of a particular tooth to the status of other teeth. RESULTS: The results revealed distinct patterns of dependence. Common patterns included either third molars and second premolars missing in all quadrants and/or combinations of these two tooth types and the incisors. The gender of the patient and the severity of hypodontia did not have a significant effect on patterns of tooth type absence. If a tooth was missing, the odds ratios that the same tooth type in the horizontally or vertically opposite quadrant was also missing were some 20 times higher than if the tooth was present. For diagonally opposite quadrants the dependence was much weaker. CONCLUSIONS: These statistical techniques are valuable for increasing understanding of hypodontia and identifying groups of patients for genetic studies.

Incisor crown shape and crowding.

The aim of this study was to investigate correlations between the shape of mandibular incisor crowns and crowding. Study models of 50 untreated white subjects (25 men and 25 women) with varying degrees of crowding were studied. The lower incisors were sectioned and imaged at the contact point and midpoint levels, and the mesiodistal width was measured. Crowding was quantified by using both Little's irregularity index and anterior tooth size-arch length discrepancy. The ratio of the 2 mesiodistal widths was correlated with Little's irregularity index and anterior tooth size-arch length discrepancy. In the women, the ratio showed correlations of 0.52 (P =.008) and 0.55 (P =.005) for central and lateral incisors, respectively, with anterior tooth size-arch length discrepancy. No predictors of lower incisor crowding could be established from mandibular incisor crown shape in this study.

Novel missense mutations and a 288-bp exonic insertion in PAX9 in families with autosomal dominant hypodontia.

We describe the molecular analysis of three families with hypodontia involving primarily molar teeth and report two novel mutational mechanisms. Linkage analysis of two large families revealed that the hypodontia was linked to the PAX9 locus. These two families revealed missense mutations consisting of a glutamic acid substitution for lysine and a proline substitution for leucine within the paired domain of PAX9. A pair of identical twins affected with hypodontia in a third family demonstrated a 288-bp insertion within exon 2 that resulted in a putative frameshift mutation and a premature stop codon. The insertion was associated with the loss of 7-bp from exon 2. A block of 256-bp of sequence within the insertion was completely identical to downstream sequence from the second intron of the PAX9 gene. These studies extend the spectrum of mutations in PAX9 associated with hypodontia to include heretofore undescribed categories, including missense mutations.