New Bone Growth into Surgical Bone Defects Grafted with a Novel Cultivated Coral Graft: A Histomorphometric Study in Rabbit Calvarias.

Natural Coral Particles (NCPs) are a suitable scaffold material for Guided Bone Regeneration (GBR) procedures; it combines the placement of a bone substitute supporting a barrier membrane. Due to increasing sea pollution and the declarations of endangered coral species (KYOTO 1997), they are no longer suitable for the medical industry. Novel domestic corals have been grown under controlled conditions to produce cultivated coral graft (CCG) material. This study aimed to evaluate a new CCG in an in vivo experimental GBR procedure. The calvarias of 8 rabbits were surgically exposed, and circular defects 8 mm in diameter were prepared. One defect was filled with CCG particles (experimental group); the contralateral defect (control group) was spontaneously filled by blood clot. The defects were covered with a collagen membrane. Animals were euthanized after 8 weeks. Histological observations of the defects showed similar bone growth patterns in both experimental and control osteotomies. In the experimental defects, no traces of coral particles were observed. Histometric analysis showed denser bone in the pristine zone (65-66%) than in the peripheral zone for both the control (50%) and experimental defects (31%) (P= NS). The new bone percentage was reduced from the peripheral zone toward the middle and the center of the defect (31%, 32% and 27%, respectively) as the distance from the peripheral pristine bone borders increased. The existing data support the complete degradation of CCG as space-maintaining scaffold for GBR procedures.

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.

A novel micro-CT analysis for evaluating the regenerative potential of bone augmentation xenografts in rabbit calvarias.

Guided Bone Regeneration is a common procedure, yet, as new grafting materials are being introduced into the market, a reliable evaluation method is required. Critical size defect in animal models provides an accurate simulation, followed by histological sections to evaluate the new bone formation. However, histology is destructive, two-dimensional and technique-sensitive. In this study we developed a novel volumetric Micro-CT analysis to quantify new bone formation characteristics. Eight adult female New Zealand white rabbits were subjected to calvarial critical-size defects. Four 8 mm in diameter circular defects were preformed in each animal, to allow random allocation of four treatment modalities. All calvarias were scanned using Micro-CT. Each defect was segmented into four equal parts: pristine bone, outer, middle, and inner. Amira software (v. 6.3, www.fei.com ) was used to calculate the new bone volume in each region and compare it to that of the pristine bone. All grafting materials demonstrated that new bone formation decreased as it moved inward. Only the inner region differed across grafting materials (p = 0.001). The new Micro-CT analysis allowed us to divide each defect into 3D regions providing better understanding of the bone formation process. Amongst the various advantages of the Micro-CT, it enables us to quantify the graft materials and the newly formed bone independently, and to describe the defect morphology in 3D (bi- vs. uni-cortical defects). Providing an insight into the inner region of the defect can better predict the regenerative potential of the bone augmentation graft material. Therefore, the suggested Micro-CT analysis is beneficial for further developing of clinical approaches.

New Bone Growth into Surgical Bone Defects Grafted with a Novel Cultivated Coral Graft: A Histomorphometric Study in Rabbit Calvarias.

Natural Coral Particles (NCPs) are a suitable scaffold material for Guided Bone Regeneration (GBR) procedures; it combines the placement of a bone substitute supporting a barrier membrane. Due to increasing sea pollution and the declarations of endangered coral species (KYOTO 1997), they are no longer suitable for the medical industry. Novel domestic corals have been grown under controlled conditions to produce cultivated coral graft (CCG) material. This study aimed to evaluate a new CCG in an in vivo experimental GBR procedure. The calvarias of 8 rabbits were surgically exposed, and circular defects 8 mm in diameter were prepared. One defect was filled with CCG particles (experimental group); the contralateral defect (control group) was spontaneously filled by blood clot. The defects were covered with a collagen membrane. Animals were euthanized after 8 weeks. Histological observations of the defects showed similar bone growth patterns in both experimental and control osteotomies. In the experimental defects, no traces of coral particles were observed. Histometric analysis showed denser bone in the pristine zone (65-66%) than in the peripheral zone for both the control (50%) and experimental defects (31%) (P= NS). The new bone percentage was reduced from the peripheral zone toward the middle and the center of the defect (31%, 32% and 27%, respectively) as the distance from the peripheral pristine bone borders increased. The existing data support the complete degradation of CCG as space-maintaining scaffold for GBR procedures.

A new method for sex estimation based on femoral cross-sectional geometry measurements and its validation using recent and ancient populations.

Estimating sex is a fundamental task in biological and forensic anthropology. This study aimed to develop new methods for sex estimation based on femoral cross-sectional geometry (CSG) variables and to test their applicability in recent and ancient assemblages. The sample was divided into a study group (living individuals, N = 124) for creating sex prediction equations and two test groups: living individuals (N = 31) and prehistoric individuals (N = 34). The prehistoric sample was divided into three subgroups according to subsistence strategy (hunter-gatherers, early farmers that also hunted, and farmers and herders). Femoral CSG variables (size, strength, and shape) were measured from CT images using dedicated software. Discriminant functions for sex estimation were calculated for various bone completeness scenarios and validated using the test groups. Size and strength parameters were sexually dimorphic, while shape was not. Discriminant functions for sex estimation produced success rates in the living sample between 83.9 and 93.5%; the distal shaft yielded the highest results. Success rates were lower among the prehistoric test sample, with better results (83.3%) for the mid-Holocene population (farmers and herders) than for earlier groups (e.g., hunter-gatherers; < 60%). These results were compared with those obtained using other methods for sex estimation based on various skeletal elements. This study provides new, reliable, and simple methods with high success rates for sex estimation based on femoral CSG variables obtained automatically from CT images. Discriminant functions were created for various conditions of femoral completeness. However, these functions should be used carefully in past populations from different settings.

Evolutionary roots of the risk of hip fracture in humans.

The transition to bipedal locomotion was a fundamental milestone in human evolution. Consequently, the human skeleton underwent substantial morphological adaptations. These adaptations are responsible for many of today's common physical impairments, including hip fractures. This study aims to reveal the morphological changes in the proximal femur, which increase the risk of intracapsular hip fractures in present-day populations. Our sample includes chimpanzees, early hominins, early Homo Neanderthals, as well as prehistoric and recent humans. Using Geometric Morphometric methods, we demonstrate differences in the proximal femur shape between hominids and populations that practiced different lifestyles. We show that the proximal femur morphology is a risk factor for intracapsular hip fracture independent of osteoporosis. Changes in the proximal femur, such as the shortening of the femoral neck and an increased anterolateral expansion of the greater trochanter, are associated with an increased risk for intracapsular hip fractures. We conclude that intracapsular hip fractures are a trade-off for efficient bipedal walking in humans, and their risk is exacerbated by reduced physical activity.

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.

Response to Comment on "A Middle Pleistocene Homo from Nesher Ramla, Israel".

Marom and Rak claim, on the basis of a few mandibular features, that the Nesher Ramla (NR) Homo is a Neanderthal. Their comments lack substance and contribute little to the debate surrounding the evolution of Middle Pleistocene Homo. Limitations and preconceptions in their study prevented them from achieving resolution beyond a dichotomous interpretation of the NR as either a Neanderthal or a modern human.

Secondary Dentin Formation Mechanism: The Effect of Attrition.

Human dentin consists of a primary layer produced during tooth formation in early childhood and a second layer which first forms upon tooth eruption and continues throughout life, termed secondary dentin (SD). The effect of attrition on SD formation was considered to be confined to the area subjacent to attrition facets. However, due to a lack of three-dimensional methodologies to demonstrate the structure of the SD, this association could not be determined. Therefore, in the current study, we aimed to explore the thickening pattern of the SD in relation to the amount of occlusal and interproximal attrition. A total of 30 premolars (50-60 years of age) with varying attrition rates were evaluated using micro-computerized tomography. The results revealed thickening of the SD below the cementoenamel junction (CEJ), mostly in the mesial and distal aspects of the root (p < 0.05). The pattern of thickening under the tooth cervix, rather than in proximity to attrition facets, was consistent regardless of the attrition level. The amount of SD thickening mildly correlated with occlusal attrition (r = 0.577, p < 0.05) and not with interproximal attrition. The thickening of the SD below the CEJ coincided with previous finite element models, suggesting that this area is mostly subjected to stress due to occlusal loadings. Therefore, we suggest that the SD formation might serve as a compensatory mechanism aimed to strengthen tooth structure against deflection caused by mechanical loading. Our study suggests that occlusal forces may play a significant role in SD formation.

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.

Effects of different irrigation protocols on dentin surfaces as revealed through quantitative 3D surface texture analysis.

The combination of ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl) has been advocated as an effective irrigation methodology to remove organic and inorganic matter in root canal therapy. Yet, it was suggested that both solutions might lead to structural changes of the dentinal wall surface, depending on the order of application which might affect sealer mechanical retention. This study aims to evaluate the effect of different irrigating protocols on dentin surface roughness using quantitative 3D surface texture analysis. Data stems from 150 human root dentin sections, divided into five groups, each prepared according to one of the following protocols: Negative control; 17% EDTA; 17% EDTA followed by 5.25% NaOCl; 5.25% NaOCl; and 5.25% NaOCl followed by 17% EDTA. Each dentin sample was examined for its three-dimensional surface texture using a high-resolution confocal disc-scanning measuring system. EDTA 17% and the combined EDTA 17% with NaOCl 5.25% showed considerably higher roughness properties compared to the control and to NaOCl 5.25% alone. However, the irrigation sequence did not affect the dentin roughness properties. Therefore, mechanical retention is probably not dependent upon the selection of irrigation protocol sequence.

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.

Suggested Case of Langerhans Cell Histiocytosis in a Cretaceous dinosaur.

Susceptibility to diseases is common to humans and dinosaurs. Since much of the biological history of every living creature is shaped by its diseases, recognizing them in fossilized bone can furnish us with important information on dinosaurs' physiology and anatomy, as well as on their daily activities and surrounding environment. In the present study, we examined the vertebrae of two humans from skeletal collections with Langerhans Cell Histiocytosis (LCH), a benign osteolytic tumor-like disorder involving mainly the skeleton; they were diagnosed in life, along with two hadrosaur vertebrae with an apparent lesion. Macroscopic and microscopic analyses of the hadrosaur vertebrae were compared to human LCH and to other pathologies observed via an extensive pathological survey of a human skeletal collection, as well as a three-dimensional reconstruction of the lesion and its associated blood vessels from a µCT scan. The hadrosaur pathology findings were indistinguishable from those of humans with LCH, supporting that diagnosis. This report suggests that hadrosaurids had suffered from larger variety of pathologies than previously reported. Furthermore, it seems that LCH may be independent of phylogeny.

Changes in human mandibular shape during the Terminal Pleistocene-Holocene Levant.

The transition to food production, exploitation of 'secondary' products (e.g., milk), and advances in cookware technology have affected all aspects of human life. The aim of the present study was to follow changes in mandibular form and shape throughout the terminal Pleistocene-Holocene Levant. The hemimandibles of four populations were included in this study: Natufian hunter-gatherers (n = 10), Pre-pottery Neolithic early farmers (n = 6), Chalcolithic farmers (n = 9), Roman-Byzantine (n = 16), and modern (n = 63) populations. A surface mesh of each mandible was reconstructed from CT or surface scans. Changes in mandibular form and shape were studied using the Procrustes-based geometric morphometrics method. Univariate and multivariate analyses were carried out to examine differences in size and shape between the studied populations. Our results reveal considerable temporal changes in mandibular shape throughout the Holocene Levant, mainly between the pre-agricultural population (the Natufian) and the succeeding ones, and between the post-industrial (the Modern) and the pre-industrial populations. A tendency for a reduction in mandibular size was identified between the pre-agricultural population and the farmers. Most regions of the mandible underwent shape changes. In conclusion, substantial changes in mandibular shape occurred throughout the Holocene Levant, especially following the agricultural revolution. These changes can be explained by the "masticatory-functional hypothesis".

Human mandibular shape is associated with masticatory muscle force.

Understanding how and to what extent forces applied to the mandible by the masticatory muscles influence its form, is of considerable importance from clinical, anthropological and evolutionary perspectives. This study investigates these questions. Head CT scans of 382 adults were utilized to measure masseter and temporalis muscle cross-sectional areas (CSA) as a surrogate for muscle force, and 17 mandibular anthropometric measurements. Sixty-two mandibles of young individuals (20-40 years) whose scans were without artefacts (e.g., due to tooth filling) were segmented and landmarked for geometric morphometric analysis. The association between shape and muscle CSA (controlled for size) was assessed using two-block partial least squares analysis. Correlations were computed between mandibular variables and muscle CSAs (all controlled for size). A significant association was found between mandibular shape and muscle CSAs, i.e. larger CSAs are associated with a wider more trapezoidal ramus, more massive coronoid, more rectangular body and a more curved basal arch. Linear measurements yielded low correlations with muscle CSAs. In conclusion, this study demonstrates an association between mandibular muscle force and mandibular shape, which is not as readily identified from linear measurements. Retrodiction of masticatory muscle force and so of mandibular loading is therefore best based on overall mandibular shape.