Primary tooth size asymmetry in twins and singletons.

OBJECTIVES: To explore asymmetry values of antimeric deciduous tooth crown dimensions in three types of twins: monozygotic (MZ), dizygotic same-sex (DZ) and opposite-sex (OS) vs. single-born controls. SETTING AND SAMPLE POPULATION: Mesiodistal and labio-lingual crown dimensions of second deciduous molars and mesiodistal canine and first molar crown dimensions of 2159 children at 6-12 years of age were evaluated, originating from the US cross-sectional Collaborative Perinatal Study from the 1970s, including altogether MZ (n = 28), DZ same-sex (n = 33) and OS (n = 39) pairs. Single born (n = 1959) were used as controls. MATERIAL AND METHODS: Dental casts were measured for comparison of variance relationships calculated from antimeric teeth, exhibiting fluctuating (FA), and directional (DA) asymmetry using anova. RESULTS: Significant differences appeared in MZ and OS girls in DA of deciduous canines, which gain size in the first and second trimester, and deciduous second molars, which finally stop crown growth during the early post-natal period. Significantly, increased FA values appeared for lower deciduous canines and second molars, indicating greatest environmental stress in OS girls, MZ girls and DZ boys. Twin girls had more fluctuating and directional crown asymmetry than twin boys, but in some dimensions, the twins were more symmetric than controls. CONCLUSIONS: Transmembrane hormonal influence between opposite-sex twins, and late gestational stress factors, caused by placental malfunction and/or monochorionicity, may be involved in asymmetric growth of antimers, during critical periods of crown size gain.

Construction of a cDNA library for miniature pig mandibular deciduous molars.

BACKGROUND: The miniature pig provides an excellent experimental model for tooth morphogenesis because its diphyodont and heterodont dentition resembles that of humans. However, little information is available on the process of tooth development or the exact molecular mechanisms controlling tooth development in miniature pigs or humans. Thus, the analysis of gene expression related to each stage of tooth development is very important. RESULTS: In our study, after serial sections were made, the development of the crown of the miniature pigs' mandibular deciduous molar could be divided into five main phases: dental lamina stage (E33-E35), bud stage (E35-E40), cap stage (E40-E50), early bell stage (E50-E60), and late bell stage (E60-E65). Total RNA was isolated from the tooth germ of miniature pig embryos at E35, E45, E50, and E60, and a cDNA library was constructed. Then, we identified cDNA sequences on a large scale screen for cDNA profiles in the developing mandibular deciduous molars (E35, E45, E50, and E60) of miniature pigs using Illumina Solexa deep sequencing. Microarray assay was used to detect the expression of genes. Lastly, through Unigene sequence analysis and cDNA expression pattern analysis at E45 and E60, we found that 12 up-regulated and 15 down-regulated genes during the four periods are highly conserved genes homologous with known Homo sapiens genes. Furthermore, there were 6 down-regulated and 2 up-regulated genes in the miniature pig that were highly homologous to Homo sapiens genes compared with those in the mouse. CONCLUSION: Our results not only identify the specific transcriptome and cDNA profile in developing mandibular deciduous molars of the miniature pig, but also provide useful information for investigating the molecular mechanism of tooth development in the miniature pig.

Precise chronology of differentiation of developing human primary dentition.

While correlation of developmental stage with embryonic age of the human primary dentition has been well documented, the available information regarding the differentiation timing of the primary teeth was largely based on the observation of initial mineralization and varies significantly. In this study, we aimed to document precise differentiation timing of the developing human primary dentition. We systematically examined the expression of odontogenic differentiation markers along with the formation of mineralized tissue in each developing maxillary and mandibular teeth from human embryos with well-defined embryonic age. We show that, despite that all primary teeth initiate development at the same time, odontogenic differentiation begins in the maxillary incisors at the 15th week and in the mandibular incisors at the 16th week of gestation, followed by the canine, the first primary premolar, and the second primary premolar at a week interval sequentially. Despite that the mandibular primary incisors erupt earlier than the maxillary incisors, this distal to proximal sequential differentiation of the human primary dentition coincides in general with the sequence of tooth eruption. Our results provide an accurate chronology of odontogenic differentiation of the developing human primary dentition, which could be used as reference for future studies of human tooth development.

Fetal age estimation using MSCT scans of deciduous tooth germs.

Evaluation of fetal age is an essential element in many fields such as anthropology, odontology, paleopathology, and forensic sciences. This study examines the correlation between fetal age, femoral diaphyseal length (considered as the gold standard), and deciduous tooth germs of fetuses aged 22 to 40 weeks amenorrhea (WA) based on computed tomography (MSCT) reconstructions. Qualitative and quantitative studies of femoral and deciduous tooth germ lengths were performed on 81 fetuses (39 females and 42 males). R software was used for statistical analyses. Intra-observer and inter-observer variabilities and the interclass correlation coefficient (ICC) were calculated. Correlation coefficients (R (2)) and linear regression equations were calculated. Intra- and inter-observer variabilities were very satisfactory (intra-observer ICC ≥ 0.96, inter-observer ICC ≥ 0.95). Femoral length was significantly correlated with age (R (2) = 0.9). The correlation coefficient between age and height, width, and dental volume was R (2) ≥ 0.73. Tooth germs were good indicators of fetal age. Our method appears to be reliable and reproducible, and the results of this study agreed with those of the literature. The dental formula provided a precise estimation of fetal age between 25 and 32 WA. Tooth germs were reliable indicators of fetal age, and multislice computed tomography was shown to be an innovative and reliable technology for this purpose.

Advances in the identification of deciduous molar tooth germs.

In forensic anthropology, correct identification of human deciduous teeth is of paramount importance for age-at-death estimation and relies on detailed anatomical descriptions. Yet literature is scarce on indications: details on the morphology of molar tooth germs of fetuses and newborns, developing from multiple mineralized centers that will eventually coalesce, are scant. This paper presents new anatomical elements for practitioners to identify human molar tooth germs at early developmental stages. 126 deciduous molars from 22 modern skeletons of fetuses and newborns (with a known age-at-death ranging between 0 days and 2 months and 21 days postnatal), without reported or observed dental pathological signs, were selected from the Collezione Antropologica LABANOF (CAL) documented skeletal collection. Gross anatomical descriptions of the morphology and configuration of the centers were provided, considering the number of mineralized centers, the shape and the outline of the occlusal plane at different stages. Three different developmental stages were observed in the maxillary first and second molar and the mandibular first molar, whereas in the mandibular second molar four stages were observed. For each stage, we provide additional detailed morphological descriptions, sketches outlining the shape of the tooth germ, and a picture of the tooth; also, indications for siding the teeth are presented. This information can be used by forensic anthropologists and odontologists for a proper identification when tooth germs are not found in anatomical connection within the dental sockets. Further analyses that encompass more age groups on a larger sample would allow to map the entire crown development of deciduous molars.

The Spy VI child: a newly discovered Neandertal infant.

Spy cave (Jemeppe-sur-Sambre, Belgium) is reputed for the two adult Neandertal individuals discovered in situ in 1886. Recent reassessment of the Spy collections has allowed direct radiocarbon dating of these individuals. The sorting of all of the faunal collections has also led to the discovery of the remains of a Neandertal child, Spy VI. This individual is represented by two mandibular corpus fragments. The left fragment is the most complete and both sides preserve the mental foramen. Four deciduous teeth are associated with these mandibular remains: three incisors and one canine. The lower left canine (Spy 645a) conjoins with the corresponding alveolar socket in the left part of the mandible. Following extant standards, the developmental stage of the preserved teeth indicate an age at death of about one and a half years. In addition to performing a classical morphometric comparative study of the mandible and teeth, we have evaluated the dental tissue proportions using high-resolution microtomographic techniques. Our results show that Spy VI generally falls within the Neandertal range of variation. However, this specimen also exhibits particular traits, notably in the dental internal structural organization, which reveals that variation in the immature Neandertal variation is larger than what was variation currently represented by the available fossil record. These observations demonstrate the need for investigating the frequency and expression of immature Neandertal traits in fossil anterior teeth, as well as their temporal and geographic variation. Direct radiocarbon dating of the Spy VI specimen has been conducted in two different laboratories. The results of Spy VI confirm the age previously determined for the two adults, making the Spy Neandertal remains the youngest ever directly dated in northwest Europe.

Early morphogenesis of heterodont dentition in minipigs.

The minipig provides an excellent experimental model for tooth morphogenesis because its diphyodont and heterodont dentition resemble that of humans. However, little information is available on the processes of tooth development in the pig. The purpose of this study was to classify the early stages of odontogenesis in minipigs from the initiation of deciduous dentition to the late bell stage when the successional dental lamina begins to develop. To analyze the initiation of teeth anlagens and the structural changes of dental lamina, a three-dimensional (3D) analysis was performed. At the earliest stage, 3D reconstruction revealed a continuous dental lamina along the length of the jaw. Later, the dental lamina exhibited remarkable differences in depth, and the interdental lamina was shorter. The dental lamina grew into the mesenchyme in the lingual direction, and its inclined growth was underlined by asymmetrical cell proliferation. After the primary tooth germ reached the late bell stage, the dental lamina began to disintegrate and fragmentize. Some cells disappeared during the process of lamina degradation, while others remained in small islands known as epithelial pearls. The minipig can therefore, inter alia, be used as a model organism to study the fate of epithelial pearls from their initiation to their contribution to pathological structures, primarily because of the clinical significance of these epithelial rests.

Quantitative proteomic analysis of deciduous molars during cap to bell transition in miniature pig.

Taking advantage of genetic manipulation tools and accessibility, almost all molecular knowledge on vertebrate tooth development was obtained from rodent models that only have one dentition in their entire lives. Whether the tooth development in other vertebrates such as swine or human follows the same rules remains elusive. Rodent dentitions differ considerably from human dentitions, therefore limiting the application of knowledge from rodent tooth to human tooth. Signal-mediated communication between cells and complex gene and protein regulatory networks are key components of tooth development. By combining isobaric tandem mass tag (TMT) labeling with liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) technology, we constructed the proteomic profile of deciduous molars at embryonic days 40 and 50 in miniature pig (Sus scrofa). During the ten days of prenatal development of the miniature pig, the morphology of the lower deciduous molar moves from the early cap to the bell stage. Thus, we identified proteins that are associated with these developing stages and identified differentially regulated proteins (DRPs) that are potential or novel drivers of tooth morphogenesis. Three candidate proteins were validated via qRT-PCR, western blotting analysis, and the location of those proteins in tooth germ were observed by immunohistochemical staining. Multiple signaling pathways and protein interaction network revealed potential mechanisms of early tooth programming in a large mammal. Bioinformatic analysis also showed that cross interaction of Wnt and Sonic hedgehog pathways may play a key role in deciduous development during cap to bell transition in miniature pig. SIGNIFICANCE: We performed the most comprehensive study of the whole tooth germ proteome in mammals to date. The high-throughput proteomic analysis identifies differentially regulated proteins and pathways that will help elucidate the mechanisms of tooth development.

Prenatal growth acceleration in maxillary deciduous canines of children with Down syndrome: histological and chemical composition study.

Previous studies have reported that the abnormal development of the second deciduous molar in Down syndrome and cerebral palsy begins before birth. In view of these results we have turned our attention to the earlier stages of dental development in utero, represented by the primary canine, in order to see if we can identify more precisely the origin and timing of developmental insults in these conditions. The study was carried out on exfoliated or extracted maxillary primary canines of children with Down syndrome (DS) and cerebral palsy (CP) and they were compared to a control group of children with no adverse medical history. Thin sections were made through the mid-sagittal bucco-palatinal axis. Using a light microscope, the width of prenatal enamel and postnatal enamel, defined by the neonatal line was measured on each section at a standardized location. The chemical composition of the enamel was then measured at three different locations using an energy dispersive spectrophotometer (ESR) in a high vacuum mode. The total enamel width in DS and controls was similar and greater than that of CP canines. Significantly more enamel was laid down prenatally in DS teeth than in controls or CP and it was more highly mineralized. These results for DS canines differ from those previously published for the later developing second primary molars. They support the hypothesis of accelerated growth in the early stages of intra-uterine development, prior to the establishment of reduced growth trajectories in the later stages. The results for CP teeth showed that more prenatal enamel was laid down prenatally than in controls. Mineralization in CP was poor during the first two trimesters and improved significantly during the last trimester. While this approach is retrospective, we propose that it may aid in identifying the onset of developmental anomalies of unknown etiology that are expressed in later life.

Remarks on the morphology of the human temporomandibular joint in the fetal period.

Skeletons of human fetuses of different ages allow the study of the considerable transformations of the mandibular joint and the mandible in this relative short stage of life. The condyle is anchored in the mandible by a conical process. The tip of this cone extends to the anLage of the 2nd milk molar. The conical process can be recognized macroscopically up to newborn age. It can also be identified by modern imaging methods. The cone can be well distinguished from the surrounding bone of the mandible. The mandibular joints of fetuses in the 31st, the 32nd, the 39th week and of a newborn were dissected, removed, and histologically investigated. The conical process of the condyle was clearly observed. The cartilage on the condyle is characterised by a layered structure which is typical for a center of growth. In the cartilage of young fetuses, blood vessels were found, reaching from the trabecular bone to the articular space. At this stage of life, blood vessels are also present in the central part of the discus articularis.

Insular dentin formation pattern in human odontogenesis in relation to the scalloped dentino-enamel junction.

This study is a first report on the modality of early dentin formation in respect to the scalloped pattern of the dentino-enamel junction (DEJ). We applied scanning electron microscopy (SEM), transmission electron microscopy (TEM), histological serial sections, and three-dimensional (3D) reconstructions. TEM and SEM showed scallops and secondary scallops on the DEJ of deciduous dental primordia and on deciduous teeth with the enamel cap removed. This peculiar outline of the DEJ requires a specific dentin formation pattern; histological sections showed that dentin formation began at the brims of the scallops, seen as triangular spikes in serial sections. The dentin formation front was not uniform; instead, it was characterized by multiple, insular forming centers, as revealed by our 3D reconstructions. As thicker dentin layers formed, the islands became confluent. Factors are discussed, which may lead to crimpling of the inner enamel epithelium, and maintained as the scalloped pattern of the DEJ develops. Signaling patterns in accordance with the insular dentin formation are unknown so far.

New regression formula to estimate the prenatal crown formation time of human deciduous central incisors derived from a Roman Imperial sample (Velia, Salerno, Italy, I-II cent. CE).

The characterization and quantification of human dental enamel microstructure, in both permanent and deciduous teeth, allows us to document crucial growth parameters and to identify stressful events, thus contributing to the reconstruction of the past life history of an individual. Most studies to date have focused on the more accessible post-natal portion of the deciduous dental enamel, even though the analysis of prenatal enamel is pivotal in understanding fetal growth, and reveals information about the mother's health status during pregnancy. This contribution reports new data describing the prenatal enamel development of 18 central deciduous incisors from the Imperial Roman necropolis of Velia (I-II century CE, Salerno, Italy). Histomorphometrical analysis was performed to collect data on prenatal crown formation times, daily secretion rates and enamel extension rates. Results for the Velia sample allowed us to derive a new regression formula, using a robust statistical approach, that describes the average rates of deciduous enamel formation. This can now be used as a reference for pre-industrial populations. The same regression formula, even when daily incremental markings are difficult to visualize, may provide a clue to predicting the proportion of infants born full term and pre-term in an archaeological series.

Prevalence of dental anomalies in deciduous dentition and its association with succedaneous dentition: A cross-sectional study of 4180 South Indian children.

OBJECTIVE: The objective of this study was to estimate the prevalence of dental anomalies in primary dentition of Indian population. MATERIALS AND METHODS: This cross-sectional study was conducted on 4180 children in the age of 2-6 years. Anomalies were classified based on Kreiborg criteria. The term "double tooth" was used to avoid misinterpretation between gemination. and fusion. Patients having radiographs were also examined for associated dental anomalies in permanent dentition. The occurrence and gender prevalence were evaluated using descriptive statistics. RESULTS: About 95. (2.27%) children exhibited at least one dental anomaly. Thirty.seven children showed 51 missing teeth. (0.88%), mostly in lower right incisors with a statistically significant difference between arches. (P = 0.0056) Nine children. (0.21%) had supernumerary teeth commonly in the right maxilla. Two cases of oligodontia. (0.04%) and talon cusps. (0.04%) and one case of triple tooth. (0.02%) were observed. Forty children. (0.95%) had 43 double teeth mostly in the right mandible with a statistically significant difference between the arches. (P = 0.0105). No significant difference was observed based on gender and arch, but they were statistically significant between the right and left sides. (P = 0.018). Among the children with radiographs available, 45% showed anomalies in the succedaneous dentition. CONCLUSIONS: The prevalence rates of children with double tooth, hypodontia, and hyperdontia in our study are 0.95%, 0.88%, and 0.21%, respectively. The overall prevalence rate of anomalies among boys was higher than girls.

Origin of the deciduous upper lateral incisor and its clinical aspects.

The upper lateral incisor in humans is often affected by dental anomalies that might be explained developmentally. To address this question, we investigated the origin of the deciduous upper lateral incisor (i2) in normal human embryos at prenatal weeks 6-8. We used serial frontal histological sections and computer-aided 3D reconstructions. At embryonic days 40-42, two thickenings of the dental epithelia in an "end-to-end" orientation were separated by a groove at the former fusion site of the medial nasal and maxillary processes. Later, these dental epithelia fused, forming a continuous dental lamina. At the fusion site, i2 started to develop. The fusion line was detectable on the i2 germ until the 8th prenatal week. The composite origin of the i2 may be associated with its developmental vulnerability. From a clinical aspect, a supernumerary i2 might be a form of cleft caused by a non-fusion of the dental epithelia.

Microstructure and chemical composition of primary teeth in children with Down syndrome and cerebral palsy.

This study was designed to test the hypothesis that prenatal growth insults leave permanent signs in the developing primary teeth that can be identified in later life. To test this hypothesis we examined exfoliated and extracted lower second primary molars of children with Down syndrome (DS) and cerebral palsy (CP). Teeth of children with no adverse medical history were used as a control group. Informed consent of parents and children was obtained in all cases. On each tooth two thin sections were cut, one bisecting the mesial cusps and one bisecting the distal cusps. Using a light microscope, the width of prenatal enamel and postnatal enamel was measured on each section at standardized locations from the dentin-enamel junction (DEJ) with the neonatal line used to distinguish between prenatal and postnatal enamel. Chemical analysis of each section was carried out using an energy dispersive spectrophotometer (ESR). The Ca/P ratios of enamel and dentin for each cusp were calculated and intercusp and intergroup differences analysed using non-parametric statistical tests. The results showed that significantly less enamel was laid down prenatally in DS and CP teeth than in the control group and that the enamel of the mesial cusps in these groups was less highly mineralised than that of the controls. The results also showed that in DS teeth growth and mineralisation of all cusps was affected. Based on these findings we propose that analysis of exfoliated deciduous teeth in developmentally challenged children may help in identifying the onset and severity of growth insults in utero and its impact on later development.

Runx2 (Cbfa1) inhibits Shh signaling in the lower but not upper molars of mouse embryos and prevents the budding of putative successional teeth.

Heterozygous mutations in the RUNX2 (CBFA1) gene cause cleidocranial dysplasia, characterized by multiple supernumerary teeth. This suggests that Runx2 inhibits successional tooth formation. However, in Runx2 knockout mice, molar development arrests at the late bud stage, and lower molars are more severely affected than upper ones. We have proposed that compensation by Runx3 may be involved. We compared the molar phenotypes of Runx2/Runx3 double-knockouts with those of Runx2 knockouts, but found no indication of such compensation. Shh and its mediators Ptc1, Ptc2, and Gli1 were down-regulated only in the lower but not the upper molars of Runx2 and Runx2/Runx3 knockouts. Interestingly, in front of the mutant upper molar, a prominent epithelial bud protruded lingually with active Shh signaling. Similar buds were also present in Runx2 heterozygotes, and they may represent the extension of dental lamina for successional teeth. The results suggest that Runx2 prevents the formation of Shh-expressing buds for successional teeth.

The developmental relationship between the deciduous dentition and the oral vestibule in human embryos.

In humans, there is no consensus about the developmental relationship between the deciduous dentition and the oral vestibule separating the teeth from the lips and cheeks. The classical concept assumes that two horseshoe-shaped epithelial structures exist: the dental lamina, giving rise to single tooth primordia, and the vestibular lamina running parallel and externally to it, giving rise to the oral vestibule. The aim of this study was to investigate the development of the dental and vestibular laminae in the upper jaw and to determine their developmental relationship in humans from embryonic week 6 to 9. Although a thickening of the vestibular epithelium was always present on serial histological sections, computer-aided three-dimensional reconstructions did not show any continuous vestibular lamina. Several discontinuous epithelial structures (bulges and ridges) occurred transiently at different stages of oral vestibule development. Along the mesiodistal axis, the dental and vestibular epithelia were regionalized in parallel: in the incisive, canine, and 1st and 2nd molar regions. The vestibular ridges fused with the dental lamina distally to the deciduous canine, 1st molar and 2nd molar. These interactions between the developing teeth and vestibular structures are reminiscent of the situation in some reptiles, where single teeth are paired one-to-one with single tooth glands.

Prenatal factors associated with the neonatal line thickness in human deciduous incisors.

The neonatal line (NNL) is used to distinguish developmental events observed in enamel which occurred before and after birth. However, there are few studies reporting relationship between the characteristics of the NNL and factors affecting prenatal conditions. The aim of the study was to determine prenatal factors that may influence the NNL thickness in human deciduous teeth. The material consisted of longitudinal ground sections of 60 modern human deciduous incisors obtained from full-term healthy children with reported birth histories and prenatal factors. All teeth were sectioned in the labio-lingual plane using diamond blade (Buechler IsoMet 1000). Final specimens were observed using scanning electron microscopy at magnifications 320×. For each tooth, linear measurements of the NNL thickness were taken on its labial surface at the three levels from the cemento-enamel junction. The difference in the neonatal line thickness between tooth types and between males and females was statistically significant. A multiple regression analyses confirmed influence of two variables on the NNL thickness standardised on tooth type and the children's sex (z-score values). These variables are the taking of an antispasmodic medicine by the mother during pregnancy and the season of the child's birth. These two variables together explain nearly 17% of the variability of the NNL. Children of mothers taking a spasmolytic medicine during pregnancy were characterised by a thinner NNL compared with children whose mothers did not take such medication. Children born in summer and spring had a thinner NNL than children born in winter. These results indicate that the prenatal environment significantly contributes to the thickness of the NNL influencing the pace of reaching the post-delivery homeostasis by the newborn's organism.

Adjacency effects in developmental correlations among tooth organs in human fetuses.

This study explores whether the "distance gradient" model shown for embryonic development and postnatal tooth growth is also characteristic of the fetal period. Histologic data and ratings from 26 human fetuses show that, unlike the embryonic and postnatal periods, changes occurring in two tooth germs are significantly alike, regardless of the number of intervening teeth.

Deciduous tooth chronology in the mandible of the domestic pig.

Fetuses of known age, collected from 20 days' gestation to term, were used to characterize the chronology of deciduous tooth development within the right mandible of swine. Tooth development was first observed at 32 days' embryonic development, with the differentiation of the deciduous third molar. Bud stages for the remaining deciduous teeth differentiated within the period of 32 to 38 days of embryonic development. Although the initial histological appearance of these teeth covered a short period of time, it was apparent that each tooth continued to develop at its own rate. The deciduous second incisor and first molar reached a stage of enamel formation by the 80th and 86th day of fetal development. This is a much later stage than previously recorded for beginning enamel formation. The stages of tooth development and enamel formation for each tooth are summarized. A previous report on the distribution of the dental lamina and deciduous tooth development in the mandible of the domestic pig combined with the information presented in this report on tooth chronology provide much of the information required for future studies using the domestic pig in dental research. A fetus observed at the 74th day of development demonstrated a tooth bud for the deciduous first premolar. The development of this tooth was followed closely throughout the remainder of fetal development with the cap stage representing its most definitive form at 110 days' development. The suggested deciduous origin for this tooth could result in a reevaluation of the nomenclature for the dental formula of swine.