Craniomaxillofacial morphology in a murine model of ephrinB1 conditional deletion in osteoprogenitor cells.

OBJECTIVE: EFNB1 mutation causes craniofrontonasal dysplasia (CFND), a congenital syndrome associated with craniomaxillofacial anomalies characterised by coronal craniosynostosis, orbital hypertelorism, and midface dysplasia. The aim of this murine study was to investigate the effect of the EfnB1 conditional gene deletion in osteoprogenitor cells on the craniomaxillofacial skeletal morphology. DESIGN: The skulls of male and female mice, in which EfnB1 was deleted by Cre (a site-specific DNA recombinase) under the control of the Osterix (Osx) promoter (EfnB1OB-/-), were compared to those without EfnB1 deletion (Osx:Cre control) at two ages (4 and 8 weeks; n = 6 per group). The three-dimensional micro-computed tomography reconstructions were prepared to calculate 17 linear measurements in the cranial vault (brain box), midface and mandible. Coronal and sagittal sutures from the 8-week-old mice were also subjected to histological examination. RESULTS: EfnB1OB-/- mice displayed significantly larger cranial height, larger interorbital and nasal widths, smaller maxillary width than controls by 8 weeks (p < 0.05), but mandibular size was not significantly different (p > 0.05). Binomial testing showed significantly smaller EfnB1OB-/- skulls at 4 weeks but larger at 8 weeks (p < 0.05). Histological examination revealed increased bony fusion and fibrous connective tissue deposition at the coronal suture of EfnB1OB-/- mice compared with controls. CONCLUSIONS: Craniofacial phenotype of the murine model of EfnB1 deletion in osteoprogenitor cells partially represents the human CFND phenotype, with implications for better understanding mechanisms involved in skeletal morphogenesis and malocclusion.

Craniofacial Phenomics: Three-Dimensional Assessment of the Size and Shape of Cranial and Dentofacial Structures.

Craniofacial phenomics has opened up numerous opportunities to correlate genetic and epigenetic factors to craniofacial phenotypes in order to improve our understanding of growth and development in health and disease. Three-dimensional (3D) imaging has played a key role in advancing craniofacial phenomics by facilitating highly sensitive and specific characterizations of craniofacial and dental morphology. Here we describe the use of micro-computed tomography (micro-CT) to image the murine craniofacial complex, followed by surface reconstruction for traditional morphometric analyses. We also describe the application of geometric morphometrics, based on Generalized Procrustes Analysis, for use in human premolars. These principles are interchangeable between various vertebrate species, and between various surface imaging techniques (including micro-CT and 3D surface scanners), offering a high level of versatility and precision for extensive phenotyping of the entire craniofacial complex.

Tooth wear development in the Australian Aboriginal dentition from Yuendumu: A longitudinal study.

The analysis of dental wear, at both the microscopic and macroscopic scale, is one of the most widely used tools in archeology and anthropology to reconstruct the diet and lifestyle of past human populations. Biomechanical studies have indicated that tooth wear helps to dissipate the mechanical load over the crown surface, thus reducing the risk of tooth fracture. To date, there are only a few studies that have examined functional tooth wear variation in modern humans. Here we propose to study masticatory efficiency through the use of the Occlusal Fingerprint Analysis method, a well-developed digital approach that allows the reconstruction of the occlusal dynamics occurring during mastication. The aim of this study is to provide the first longitudinal quantitative data of molar and premolar macrowear patterns within a functional context. We examined the mixed and permanent dentition of one Australian Aboriginal child (from ages 8 to 17) from Yuendumu, using high-resolution surface scans of dental casts including both upper and lower arches. Our results suggest that the occlusal macrowear patterns of this individual did not significantly change through time. Occlusal contact parameters such as functional area, inclination and direction remain relatively unaltered throughout childhood and adolescence, indicating little change in the masticatory function of this individual. The functional tooth wear pattern in this individual did not change longitudinally indicating the degree of masticatory efficiency has most probably remained unaltered.

A functional analysis of Carabelli trait in Australian aboriginal dentition.

OBJECTIVES: Carabelli is a nonmetric dental trait variably expressed as a small pit to a prominent cusp in the maxillary molars of modern humans. Investigations on the occurrence and expression rates of this trait have been conducted extensively, tracing its origin to genetic sources. However, there remains a lack of understanding about its potential role in chewing. In this study, we examine molar macrowear with the aim of reconstructing Carabelli trait occlusal dynamics occurring during chewing. METHODS: We have examined 96 deciduous and permanent maxillary molars of children and young adults from Yuendumu, an Australian Aboriginal population that was at an early stage of transition from a nomadic and hunter-gatherer way of life to a more settled existence. We apply a well-established method, called Occlusal Fingerprint Analysis, which is a digital approach for analyzing dental macrowear allowing the reconstruction of jaw movements required to produce wear pattern specific to each tooth. RESULTS: Carabelli trait slightly enlarges the surface functional area, especially in those molars where this feature is expressed in its cuspal form and it is closer to the occlusal plane. Moreover, the highly steep contact planes would also indicate that Carabelli wear areas contribute to increasing the shearing abilities of the occluded teeth, which are particularly important when processing fibrous and tough foods. CONCLUSIONS: The macrowear analysis suggests that Carabelli trait in the Aboriginal people from Yuendumu slightly enhanced occlusion and probably played some functional role during mastication. Future biomechanical and microwear analyses could provide additional information on the mechanical adaptation of Carabelli trait in modern human dentition.

Dentinal dysplasia type 1: A 3D micro-computed tomographic study of enamel, dentine and root canal morphology.

Dentine dysplasia type 1 is a rare and complex dental anomaly. Our aim was to conduct a morphometric assessment of a dentinal dysplasia type 1c (DD1c) caries-free mandibular second molar, extracted due to symptomatic apical periodontitis. Controls consisted of five intact mandibular second molars. Micro-computed tomography analysis showed that the DD1c volume % for enamel, dentine/cementum and pulp chamber fell in the 0.36th, 99.97th and 0.09th percentiles of the control teeth (P < 0.01). It also revealed an extremely complicated root canal system in the DD1c tooth with a varying degree of dentine mineralisation and aberrant dentine deposition in the pulp chamber. A crack extending from the external tooth surface to the pulp chamber was identified as a potential site for microbial invasion. Clinical implications include preventive measures and early intervention in reversible pulpitis. Conclusion: Micro-CT imaging can be useful in establishing post-extraction diagnosis of cracks and phenomic characterisation of tooth anomalies.

Craniofacial abnormalities in a murine model of Saethre-Chotzen Syndrome.

BACKGROUND: Saethre-Chotzen Syndrome (SCS) is an autosomal dominant syndrome that occurs due to a mutation or deletion of the Twist1 gene at chromosome 7p21. Our aim was to conduct a morphometric analysis of the craniofacial features in the mouse associated with a Twist1+/- mutation. METHODS: Micro-computed imaging was conducted for the skulls of forty skeletally mature mice, equally distributed by sex (male and female) and two genotypes (Twist1+/- or murine model of SCS; and Twist1+/+ or wild-type). A morphometric analysis was carried out for eight parameters for the maxillary-zygomatico-temporal region, 10 parameters for the mandible and three parameters for teeth from three-dimensional reconstructions. RESULTS: Compared with wild-type, the murine model of SCS showed these trends: (1) maxillary-zygomatico-temporal region, significantly shorter length and width posteriorly (p<0.05), (2) mandible, significantly reduced height and width (p<0.05), and (3) teeth, significantly shorter height, shorter mesio-distal width but longer bucco-lingual width (p<0.05). In the murine model of SCS, the key morphological variations included incomplete ossification of the temporal bone and zygomatic arch, twisting and/or incomplete ossification of the palatal process of the maxilla, premaxilla and the ventral nasal concha, as well as bifid coronoid processes. CONCLUSIONS: The skeletal and dental alterations in the height, length and width provide a foundation for large-scale phenomics studies, which will improve existing knowledge of the Twist1 signalling cascade. This is relevant given the predicted shift towards minimally invasive molecular medical treatment for craniosynostosis.

Technical note: The use of 3D printing in dental anthropology collections.

OBJECTIVES: Rapid prototyping (RP) technology is becoming more affordable, faster, and is now capable of building models with a high resolution and accuracy. Due to technological limitations, 3D printing in biological anthropology has been mostly limited to museum displays and forensic reconstructions. In this study, we compared the accuracy of different 3D printers to establish whether RP can be used effectively to reproduce anthropological dental collections, potentially replacing access to oftentimes fragile and irreplaceable original material. METHODS: We digitized specimens from the Yuendumu collection of Australian Aboriginal dental casts using a high-resolution white-light scanning system and reproduced them using four different 3D printing technologies: stereolithography (SLA); fused deposition modeling (FDM); binder-jetting; and material-jetting. We compared the deviations between the original 3D surface models with 3D print scans using color maps generated from a 3D metric deviation analysis. RESULTS: The 3D printed models reproduced both the detail and discrete morphology of the scanned dental casts. The results of the metric deviation analysis demonstrate that all 3D print models were accurate, with only a few small areas of high deviations. The material-jetting and SLA printers were found to perform better than the other two printing machines. CONCLUSIONS: The quality of current commercial 3D printers has reached a good level of accuracy and detail reproduction. However, the costs and printing times limit its application to produce large sample numbers for use in most anthropological studies. Nonetheless, RP offers a viable option to preserve numerically constraint fragile skeletal and dental material in paleoanthropological collections.

Three-dimensional (3D) geometric morphometric analysis of human premolars to assess sexual dimorphism and biological ancestry in Australian populations.

OBJECTIVES: This study aimed to investigate size and shape variation of human premolars between Indigenous Australians and Australians of European ancestry, and to assess whether sex and ancestry could be differentiated between these groups using 3D geometric morphometrics. MATERIALS AND METHODS: Seventy dental casts from each group, equally subdivided by sex, were scanned using a structured-light scanner. The 3D meshes of upper and lower premolars were processed using geometric morphometric methods. Seventy-two landmarks were recorded for upper premolars and 50 landmarks for lower premolars. For each tooth type, two-way ANOVA was used to assess group differences in centroid size. Shape variations were explored using principal component analysis and visualized using 3D morphing. Two-way Procrustes ANOVA was applied to test group differences for ancestry and sex, and a "leave-one-out" discriminant function was applied to assess group assignment. RESULTS: Centroid size and shape did not display significant difference between the sexes. Centroid size was larger in Indigenous Australians for upper premolars and lower second premolars compared to the Australians of European ancestry. Significant shape variation was noted between the two ancestral groups for upper premolars and the lower first premolar. Correct group assignment of individual teeth to their ancestral groups ranged between 80.0 and 92.8% for upper premolars and 60.0 and 75.7% for lower premolars. DISCUSSION: Our findings provide evidence of significant size and shape variation in human premolars between the two ancestral groups. High classification rates based on shape analysis of upper premolars highlight potential application of geometric morphometrics in anthropological, bioarcheological and forensic contexts.

Extensive phenotyping of the orofacial and dental complex in Crouzon syndrome.

OBJECTIVES: Fibroblast growth factor receptor 2 (FGFR2) C342Y/+ mutation is a known cause of Crouzon syndrome that is characterised by craniosynostosis and midfacial hypoplasia. Our aim was to conduct extensive phenotyping of the maxillary, mandibular and dental morphology associated with this mutation. MATERIALS AND METHODS: Morphometric data were obtained from 40 mice, representing two genotypes (Crouzon and wild-type) and two sexes (males and females) (n=10 in each group). Dental analysis further categorised the first molars into the two jaws (maxillary and mandibular) (n=20 in each group). Maxillary, mandibular and dental morphology was compared by analysing 23 linear landmark-based dimensions in three-dimensional micro-computed tomography reconstructions. RESULTS: Compared with wild-type, Crouzon (FGFR2C342Y/+) maxillae were significantly shorter in maximum height, anterior and posterior lengths and middle width, but larger in posterior width (p<0.05 for height; p<0.001 for other comparisons). In the Crouzon mandible, the ascending and descending heights, effective and mandibular lengths, and intercoronoid and intercondylar widths were significantly shorter, whereas intergonial width was larger (p<0.01 for intercondylar width; p<0.001 for other comparisons). Crouzon teeth were significantly smaller mesiodistally, but larger in crown height (p<0.001 for each comparison). All Crouzon mice presented with bifid mandibular condyles and a quarter presented with expansive bone lesions in the mandibular incisor alveolus. CONCLUSIONS: Our findings of hypoplasia in all three planes in Crouzon maxillae and mandibles, together with the presence of bifid mandibular condyles and expansive bone lesions, may be relevant to maxillofacial surgery and orthodontics. Beyond skeletal effects, the FGFR2C342Y/+ mutation is now implicated in affecting tooth development. This study's skeletal phenomics data also provides baseline data against which the effect of various treatments can now be assessed.