Finite element analysis of Neanderthal and early Homo sapiens maxillary central incisor.

Neanderthal anterior teeth are very large and have a distinctive morphology characterized by robust 'shovel-shaped' crowns. These features are frequently seen as adaptive responses in dissipating heavy mechanical loads resulting from masticatory and non-masticatory activities. Although the long-standing debate surrounding this hypothesis has played a central role in paleoanthropology, is still unclear if Neanderthal anterior teeth can resist high mechanical loads or not. A novel way to answer this question is to use a multidisciplinary approach that considers together tooth architecture, dental wear and jaw movements. The aim of this study is to functionally reposition the teeth of Le Moustier 1 (a Neanderthal adolescent) and Qafzeh 9 (an early Homo sapiens adolescent) derived from wear facet mapping, occlusal fingerprint analysis and physical dental restoration methods. The restored dental arches are then used to perform finite element analysis on the left central maxillary incisor during edge-to-edge occlusion. The results show stress distribution differences between Le Moustier 1 and Qafzeh 9, with the former displaying higher tensile stress in enamel around the lingual fossa but lower concentration of stress in the lingual aspect of the root surface. These results seem to suggest that the presence of labial convexity, lingual tubercle and of a large root surface in Le Moustier 1 incisor helps in dissipating mechanical stress. The absence of these dental features in Qafzeh 9 is compensated by the presence of a thicker enamel, which helps in reducing the stress in the tooth crown.

The extracellular matrix protein agrin promotes heart regeneration in mice.

The adult mammalian heart is non-regenerative owing to the post-mitotic nature of cardiomyocytes. The neonatal mouse heart can regenerate, but only during the first week of life. Here we show that changes in the composition of the extracellular matrix during this week can affect cardiomyocyte growth and differentiation in mice. We identify agrin, a component of neonatal extracellular matrix, as required for the full regenerative capacity of neonatal mouse hearts. In vitro, recombinant agrin promotes the division of cardiomyocytes that are derived from mouse and human induced pluripotent stem cells through a mechanism that involves the disassembly of the dystrophin-glycoprotein complex, and Yap- and ERK-mediated signalling. In vivo, a single administration of agrin promotes cardiac regeneration in adult mice after myocardial infarction, although the degree of cardiomyocyte proliferation observed in this model suggests that there are additional therapeutic mechanisms. Together, our results uncover a new inducer of mammalian heart regeneration and highlight fundamental roles of the extracellular matrix in cardiac repair.

Immunomodulatory fibrous hyaluronic acid-Fmoc-diphenylalanine-based hydrogel induces bone regeneration.

AIM: To investigate the potential of an ultrashort aromatic peptide hydrogelator integrated with hyaluronic acid (HA) to serve as a scaffold for bone regeneration. MATERIALS AND METHODS: Fluorenylmethyloxycarbonyl-diphenylalanine (FmocFF)/HA hydrogel was prepared and characterized using microscopy and rheology. Osteogenic differentiation of MC3T3-E1 preosteoblasts was investigated using Alizarin red, alkaline phosphatase and calcium deposition assays. In vivo, 5-mm-diameter calvarial critical-sized defects were prepared in 20 Sprague-Dawley rats and filled with either FmocFF/HA hydrogel, deproteinized bovine bone mineral, FmocFF/Alginate hydrogel or left unfilled. Eight weeks after implantation, histology and micro-computed tomography analyses were performed. Immunohistochemistry was performed in six rats to assess the hydrogel's immunomodulatory effect. RESULTS: A nanofibrous FmocFF/HA hydrogel with a high storage modulus of 46 KPa was prepared. It supported osteogenic differentiation of MC3T3-E1 preosteoblasts and facilitated calcium deposition. In vivo, the hydrogel implantation resulted in approximately 93% bone restoration. It induced bone deposition not only around the margins, but also generated bony islets along the defect. Elongated M2 macrophages lining at the periosteum-hydrogel interface were observed 1 week after implantation. After 3 weeks, these macrophages were dispersed through the regenerating tissue surrounding the newly formed bone. CONCLUSIONS: FmocFF/HA hydrogel can serve as a cell-free, biomimetic, immunomodulatory scaffold for bone regeneration.

The Hidden Secrets of the Dental Calculus: Calibration of a Mass Spectrometry Protocol for Dental Calculus Protein Analysis.

Dental calculus is a solid deposit that forms and accumulates on the tooth surface, entrapping oral microorganisms, biomolecules, and other micro-debris found in the oral cavity. A mass spectrometry analysis of its protein content opens a vista into the subject's diet, oral flora, and even some aspects of health, thus providing new insight and expanding our knowledge of archaic cultures. Multiple experimental protocols have been proposed for the optimal extraction of proteins from dental calculus. Herein, we compared various experimental conditions in order to calibrate and validate a protocol for protein extraction. Our results show that a high concentration of acetic acid followed by mechanical crushing and sonication provided the highest protein yield, while acetone precipitation enabled the identification of more distinct proteins. We validated this protocol using archeological samples, identifying human and microbial proteins in specimens from the eighth and seventeenth centuries (approximately 250-1300 years ago). These findings demonstrate that the developed protocol is useful for studying excavated archaeological samples and that it might be utilized to explore the biohistory, dietary habits, and microbiome of archaic populations.

The dental remains from the Early Upper Paleolithic of Manot Cave, Israel.

This study presents the dental remains discovered at Manot Cave (MC), Western Galilee, Israel. The cave contains evidence for human occupation during the Early Upper Paleolithic period (46-33 ka) mainly of Early Ahmarian (∼46-42 ka) and Levantine Aurignacian (∼38-34 ka) cultural levels. Six teeth (three deciduous and three permanent) were found at the site, of which four could be thoroughly analyzed. The morphology of the teeth was qualitatively described and analyzed using traditional and geometric morphometric methods. A large comparative sample was used in order to assess the morphological affiliation of the Manot specimens with other Homo groups. The results provided equivocal signals: the upper first premolar (MC-9 P3) is probably modern human; the upper deciduous second molar (MC-10 dm2) and the upper second permanent molar (MC-8 M2) might be modern humans; the lower second deciduous molar (MC-7 dm2) might be Neanderthal. Owing to the small sample size and the almost total lack of distinctive characteristics, our outcome could not supply conclusive evidence to address the question of whether Manot Aurignacian population came from Europe or descended from the local Ahmarian population.

Early Upper Paleolithic human foot bones from Manot Cave, Israel.

The transition from the Middle Paleolithic to the Upper Paleolithic in the Levant represents a major event in human prehistory with regards to the dispersal of modern human populations. Unfortunately, the scarcity of human remains from this period has hampered our ability to study the anatomy of Upper Paleolithic populations. This study describes and examines pedal bones recovered from the Early Upper Paleolithic period at Manot Cave, Israel, from 2014 to 2017. The Manot Cave foot bones include a partial, left foot skeleton comprising a talus, a calcaneus, a cuboid, a first metatarsal, a second metatarsal, a fifth metatarsal, and a hallucal sesamoid. All these remains were found in the same archaeological unit of the cave and belong to a young adult. Shape and size comparisons with Neanderthals, Anatomically Modern Human and modern human foot bones indicate a modern human morphology. In some characteristics, however, the Manot Cave foot bones display a Neanderthal-like pattern. Notably, the Manot Cave foot is remarkable in its overall gracility. A healed traumatic injury in the second metatarsal (Lisfranc's fracture) is most likely due to a remote impact to the dorsum of the foot. This injury, its subsequent debility, and the individual's apparent recovery suggest that the members of the Manot Cave community had a supportive environment, one with mutual responsibilities among the members.

The endocast of the late Middle Paleolithic Manot 1 specimen, Western Galilee, Israel.

Studying endocasts has long allowed anthropologists to examine changes in the external topography and the overall size of the brain throughout the evolutionary history of hominins. The nearly complete calvaria of Manot 1 presents an opportunity to gain insights into the external brain morphology, vascular system, and dimensions of the brain of this late Middle Paleolithic hominin. Detailed size and shape analyses of the Manot 1 endocast indicate a modern Homo sapiens anatomy, despite the presence of some primitive features of the calvaria. Traits considered to be derived endocranial features for H. sapiens are present in Manot 1, including an elongated parietal sagittal chord with an elevated superior part of the hemisphere, a widened posterior part of the frontal lobes, a considerable development of the parietal reliefs such as the supramarginal lobules, and a slight posterior projection of the occipital lobes. These findings, together with data presented in previous studies, rule out the possibility of a direct Neanderthal ancestry for the Manot 1 hominin and instead confirm its affiliation with H. sapiens. The Manot 1 calvaria is more similar to that of later Upper Paleolithic H. sapiens than it is to the earlier Levantine populations of Skhul and Qafzeh. The late Middle Paleolithic date of Manot 1 provides an opportunity to analyze the recent developments in human cerebral morphology and organization.

Exploring directional and fluctuating asymmetry in the human palate during growth.

OBJECTIVES: Palate morphology is constantly changing throughout an individual's lifespan, yet its asymmetry during growth is still little understood. In this research, we focus on the study of palate morphology by using 3D geometric morphometric approaches to observe changes at different stages of life, and to quantify the impact of directional and fluctuating asymmetry on different areas at different growth stages. MATERIALS AND METHODS: The sample consists of 183 individuals (1-72 years) from two identified human skeletal collections of 19th and early 20th Century Italian contexts. A 3D-template of 41 (semi)landmarks was applied on digital palate models to observe morphological variation during growth. RESULTS: Asymmetrical components of the morphological structure appears multidirectional on the entire palate surface in individuals <2 years old and become oriented (opposite bilateral direction) between 2 and 6 years of age. Specifically, directional asymmetry differentially impacts palate morphology at different stages of growth. Both the anterior and posterior palate are affected by mild alterations in the first year of life, while between 2 and 6 years asymmetry is segregated in the anterior area, and moderate asymmetry affects the entire palatal surface up to 12 years of age. Our results show that stability of the masticatory system seems to be reached around 13-35 years first by females and then males. From 36 years on both sexes show similar asymmetry on the anterior area. Regarding fluctuating asymmetry, inter-individual variability is mostly visible up to 12 years of age, after which only directional trends can be clearly observed at a group level. DISCUSSION: Morphological structure appears instable during the first year of life and acquires an opposite asymmetric bilateral direction between 2 and 6 years of age. This condition has been also documented in adults; when paired with vertical alteration, anterior/posterior asymmetry seems to characterize palate morphology, which is probably due to mechanical factors during the lifespan. Fluctuating asymmetry is predominant in the first period of life due to a plausible relationship with the strength of morphological instability of the masticatory system. Directional asymmetry, on the other hand, shows that the patterning of group-level morphological change might be explained as a functional response to differential inputs (physiological forces, nutritive and non-nutritive habits, para-masticatory activity as well as the development of speech) in different growth stages. This research has implications with respect to medical and evolutionary fields. In medicine, palate morphology should be considered when planning orthodontic and surgical procedures as it could affect the outcome. As far as an evolutionary perspective is concerned the dominance of directional asymmetries in the masticatory system could provide information on dietary and cultural habits as well as pathological conditions in our ancestors.

Are chin and symphysis morphology facial type-dependent? A computed tomography-based study.

INTRODUCTION: The chin is a major determinant of the facial profile; hence, it plays a major role in orthodontics and orthognathic surgery. It is thus essential to follow and better understand its expression in different facial types. The major objectives of the current study were to characterize morphometrically the chin and symphysis and reveal their association with different facial types. METHODS: Computed tomography scans of the head and neck of 311 adults (163 males, 148 females; age range, 18-95 years) were classified into 3 facial types: short, average, and long. Height, width, projection, inclination, thickness, and area were measured on the chin and symphysis. RESULTS: The majority of the population (70%) manifested an average facial type; the other 30% were almost equally distributed between short and long facial types. The long facial type was more common among females and the short facial type among males. Chin projection, area, and size were significantly greater in short-faced patients. Chin width in males was similar for all facial types, whereas, in females, chin width was the widest in the short facial type and the narrowest in the long facial type. Symphysis height was significantly greater in long-faced patients in both sexes. The mandibular incisors' inclination relative to the mandibular plane was not significantly associated with the chin or symphysis morphology. CONCLUSIONS: Chin and symphysis morphology is facial type-dependent. Orthodontists and maxillofacial surgeons should be aware of the complex relationship between facial types and chin/symphysis size and shape when planning treatment.

Age estimation of fragmented human dental remains by secondary dentin virtual analysis.

Unlike bones, teeth are remarkably resilient and can withstand severe trauma, making age assessment based on the dentition essential for forensic analysis. Modern techniques for age estimation focus on pulp-chamber volume measurements using radiographs and computerized tomography (CT); however, these are applicable only for complete teeth (i.e., with intact crown and root). In the current study, we developed a new approach using high-resolution micro-computerized tomography (µCT) to visualize the secondary dentin (SD), an inner layer surrounding the pulp which accumulates with age, thus facilitating age estimation of fragmented and broken teeth.The growth pattern of the SD with age was analyzed for 77 lower premolars from two anthropological collections. A comparison of SD virtual segmentation and histological measurement was highly correlative (ICC = 0.95). SD was measured per volume (mm3) of a 1 mm thick slice directly below the cemento-enamel junction. Regression analysis using SD measurements increased the success rates of age estimation (82%) compared with the "gold-standard" pulp/dentin method (54%) in the range of ± 10 years. The accuracy of age estimation based on SD analysis was improved to a range of 7-8 years.The SD method thus allows age estimation with greater prediction rates and better accuracy based on only a small fragment of a tooth in a non-invasive manner. This novel methodology is easy to use, accessible, and bears implications in various fields such as forensic sciences and anthropological research.

The effect of tooth wear, age and sex on facial height assessed by soft tissue analysis.

AIM: The aim of the study was to evaluate the effects of tooth wear, age and sex on facial height measurements based on soft tissue analysis. METHODS: One hundred and twelve subjects (45 males and 67 females), 19-80 years of age (mean 43 ± 0.9 years), which were divided into three age groups, met the inclusion criteria. Tooth wear was assessed using a 5-point ordinal scale (0-4). Based on the tooth wear score, three groups were defined: mild, moderate and severe wear. Facial height was evaluated based on soft tissue measurements using frontal facial photographs. The dimension of the upper facial height (UFH) and lower facial height (LFH) and the ratio between the two (R-U-L) were measured. RESULTS: Facial height dimensions (UFH, LFH) were significantly different between the three age groups and between the three tooth wear groups. Both the UFH and LFH measurements had a weak positive correlation with age (r = .40; r = .41, respectively) and with tooth wear (r = .40; r = .41, respectively). The R-U-L showed a significant difference (P < .001) between sexes as males showed slightly lower ratio compared with females; however, the R-U-L was not dependent upon age or tooth wear. CONCLUSIONS: This study demonstrated that although facial height is affected by age and tooth wear, the proportion between the upper to lower facial segments is constant and is not affected by the same parameters. A statistically significant difference in the R-U-L relative to sexes was noted.

The cancer paradigms of mammalian regeneration: can mammals regenerate as amphibians?

Regeneration in mammals is restricted to distinct tissues and occurs mainly by expansion and maturation of resident stem cells. During regeneration, even subtle mutations in the proliferating cells may cause a detrimental effect by eliciting abnormal differentiation or malignant transformation. Indeed, cancer in mammals has been shown to arise through deregulation of stem cells maturation, which often leads to a differentiation block and cell transformation. In contrast, lower organisms such as amphibians retain a remarkable regenerative capacity in various organs, which occurs via de- and re-differentiation of mature cells. Interestingly, regenerating amphibian cells are highly resistant to oncogenic transformation. Therapeutic approaches to improve mammalian regeneration mainly include stem-cell transplantations; but, these have proved unsuccessful in non-regenerating organs such as the heart. A recently developed approach is to induce de-differentiation of mature cardiomyocytes using factors that trigger their re-entry into the cell cycle. This novel approach raises numerous questions regarding the balance between transformation and regeneration induced by de-differentiation of mature mammalian somatic cells. Can this balance be controlled artificially? Do de-differentiated cells acquire the protection mechanisms seen in regenerating cells of lower organisms? Is this model unique to the cardiac tissue, which rarely develops tumors? This review describes regeneration processes in both mammals and lower organisms and, particularly, the ability of regenerating cells to avoid transformation. By comparing the characteristics of mammalian embryonic and somatic cells, we discuss therapeutic strategies of using various cell populations for regeneration. Finally, we describe a novel cardiac regeneration approach and its implications for regenerative medicine.

Sex estimation using computed tomography of the mandible.

Sex estimation of skeletal parts is of great value even in the DNA era. When computed tomography (CT) facilities were introduced to forensic institutes, new possibilities for sex estimation emerged. The aim of this study was to develop a CT-based method for sex estimation using the mandible. Twenty-five CT-based measurements of the mandible were developed and carried out on 3D reconstructions (volume rendering) and cross sections of the lower jaw of 438 adult individuals (214 males and 224 females). Intraobserver and interobserver variances of the measurements were examined using intraclass correlation coefficient (ICC) analysis. Five discriminant functions were developed using different states of completeness of the mandible. The success rates of these equations were cross validated twice. The measurements were found to be highly reliable (for intraobserver 0.838 < ICC < 0.995 and for interobserver 0.71 < ICC < 0.996). For a complete mandible, the correct classification rate was 90.8%. For incomplete mandibles, the correct classification rates varied from 72.9 to 85.6%. Cross-validation tests yielded similar success rates, for the complete mandible 89% and for the incomplete mandible 67.5 to 89%. We concluded that CT techniques are appropriate for estimating sex based on the mandible size and shape characteristics. Suggested discriminant functions for sex estimation are given with data on the correct classification rates.

Rate and pattern of interproximal dental attrition.

Although occlusal and interproximal attrition occur because of diverse etiology and present dissimilar features, both progress with age. The objectives of this study were to reveal the rate and pattern of development of interproximal attrition facets (PAF) with age and to compare those with occlusal attrition (OA) changes. Five-hundred and ninety-four teeth were collected from 198 skulls (of adults, 20-71 yr of age). Three mandibular teeth [first premolar (P1), second premolar (P2), and first molar (M1)] were examined for PAF size and OA rate. Interproximal attrition and OA followed similar patterns of development until subjects reached 40-45 yr of age, after which they took different paths: PAF did not increase in size and were not as large as in younger groups, regardless of facet location, whereas OA continued to progress. The PAF changes with age differed between premolars and molars, unlike OA, which presented a similar rate for all teeth studied. Although OA scores presented significantly moderate correlations with age, PAF area size demonstrated low correlations with age. Low, but significant, correlations were found between the rate of OA and that of PAF. However, PAF and OA exhibited different patterns of development with age. Premolars and molars presented dissimilar development of PAF, which is probably caused by a unique attrition pattern in the molar teeth, different morphology, and force vectors.

Pattern of maxillary and mandibular proximal enamel thickness at the contact area of the permanent dentition from first molar to first molar.

INTRODUCTION: Proximal enamel thickness (PET) at the mesial and distal contact areas of the complete permanent dentition has not been previously reported. Anatomic investigation of PET is essential for interproximal reduction treatment. Our objectives were to measure the PETs of the complete maxillary and mandibular dentitions at the contact areas, to compare the PETs of adjacent teeth, and to evaluate the vertical position of each contact area. METHODS: We evaluated 720 extracted teeth; of these, 109 intact teeth were selected. The mesial and distal contact areas were demarcated, and each tooth was embedded in transparent epoxy resin. Blocks were prepared so that the 2 demarcated contact areas were exposed, and 6 measurements were taken and statistically analyzed. RESULTS: Both jaws showed the same PET pattern characterized by 5 features: PET increased progressively from incisor (0.63 mm) to first molar (1.48 mm). Per tooth, mesial and distal PET did not differ. Total maxillary (26.86 mm) and mandibular (24.52 mm) PETs were similar. Paired PETs at the interproximal interface were similar, with the exception of the lateral incisor-canine interfaces. From incisor to first molar, the contact area becomes located farther gingivally. CONCLUSIONS: The existing guideline of maximal 0.5-mm interproximal reduction per 2 adjacent teeth should be kept in the anterior region and could be increased to 1 mm in the posterior region, when an equal amount is removed.

p53 coordinates cranial neural crest cell growth and epithelial-mesenchymal transition/delamination processes.

Neural crest development involves epithelial-mesenchymal transition (EMT), during which epithelial cells are converted into individual migratory cells. Notably, the same signaling pathways regulate EMT function during both development and tumor metastasis. p53 plays multiple roles in the prevention of tumor development; however, its precise roles during embryogenesis are less clear. We have investigated the role of p53 in early cranial neural crest (CNC) development in chick and mouse embryos. In the mouse, p53 knockout embryos displayed broad craniofacial defects in skeletal, neuronal and muscle tissues. In the chick, p53 is expressed in CNC progenitors and its expression decreases with their delamination from the neural tube. Stabilization of p53 protein using a pharmacological inhibitor of its negative regulator, MDM2, resulted in reduced SNAIL2 (SLUG) and ETS1 expression, fewer migrating CNC cells and in craniofacial defects. By contrast, electroporation of a dominant-negative p53 construct increased PAX7(+) SOX9(+) CNC progenitors and EMT/delamination of CNC from the neural tube, although the migration of these cells to the periphery was impaired. Investigating the underlying molecular mechanisms revealed that p53 coordinates CNC cell growth and EMT/delamination processes by affecting cell cycle gene expression and proliferation at discrete developmental stages; disruption of these processes can lead to craniofacial defects.

Proximal attrition facets: morphometric, demographic, and aging characteristics.

Although interproximal attrition is considered to be limited in modern populations, it has important clinical implications. However, in contrast to occlusal attrition, proximal attrition receives limited scientific attention. The main purpose of the current study was to fill this void. Seven-hundred and sixty-five teeth were collected from 255 skulls of subjects 18-75 yr of age. For each individual, three mandibular teeth (the first and second premolars and the first molar) were examined for proximal attrition facets (PAFs). The results provide detailed information on the size, shape, and location of the facets according to age cohort, gender, and ethnicity. The validity of the method used to measure the facets was also examined. The major findings were as follows: PAFs are usually located on the upper half of the crown proximal aspect; in each tooth, the mesial facet is more lingually positioned and the distal facet is more buccally positioned; the majority of the facets are subrectangular in shape; the size of the facets tends to increase in an anteroposterior direction (from premolars to molars); and facet size and location are age- and sex-dependent and ethnicity-independent. It is our recommendation that dentists bear in mind that interproximal attrition is a dynamic, long-term process and needs to be considered in many clinical scenarios.

Long- and short-term effects of headgear traction with and without the maxillary second molars.

INTRODUCTION: A quantitative assessment of maxillary first molar distalization with and without the maxillary second molar (M2) was carried out. METHODS: Fifty-six cervical headgear patients undergoing fixed appliance orthodontic treatment were divided into 2 groups: before (G - M2) and after (G + M2) eruption of the maxillary second molars (ages, 11.87 ± 1.20, and 13.05 ± 1.55 years, respectively). The tightness of the dental contact point (TDCP) and the space between the second premolar and the maxillary first molar were measured at 6 levels of headgear force (0-15 N) at 3 intervals 6 months apart (T0, T1, T2). RESULTS: Relationships were found between space and TDCP, time, and presence or absence of the maxillary second molar at T1 and T2 (P <0.001). The TDCP decreased and space increased with increase in initial headgear force. An increase in initial force beyond 6 to 9 N did not significantly increase the initial maxillary first molar distalization. The G - M2 TDCP and space measurements were similar to those of G + M2 at T2 with the eruption of the maxillary second molar. From T0 to T1, maxillary first molar distalization was greater in G - M2. In comparison with our previous headgear-alone study, initial distalization with a fully bonded appliance was reduced by 4-fold. CONCLUSIONS: Headgear therapy is more effective before the eruption of the maxillary second molar. Once it erupts, the distalization pace of the maxillary first molar is reduced, but it can nevertheless be pursued at a slower pace when the maxillary second molar is present.