Changes in the Craniomaxillofacial Growth and Development of Sprague Dawley Rats Subjected to Permanent Experimental Unilateral Nasal Obstruction / Cambios en el Crecimiento y Desarrollo Cráneo Maxilo Facial de Ratas Sprague Dawley Sometidas a Obstrucción Nasal Unilateral Experimental Permanente

SUMMARY: Breathing is considered a vital function dependent on factors such as adequate permeability of the nasal route, which is linked to physiological functions, intellectual processes, and craniofacial growth. The aim of this study was to determine the changes in the craniomaxillofacial growth and bone development of Sprague Dawley rats subjected to permanent experimental unilateral nasal obstruction. Twenty-four newborn rats were used, randomized, and divided into experimental and control groups. The right nostril was obstructed, and weight, length, and Lee's index measurements were recorded at 8 and 16 weeks. Craniomandibular x-rays were taken of each animal, obtaining linear neuro- and viscerocranial measurements. Then, a biochemical analysis was performed to measure the alkaline phosphatase concentration. The results were analyzed in the SPSS software, performing a descriptive analysis, using a t-test for independent samples, comparing basal, cephalometric, and biochemical characteristics between the control and experimental groups, considering a significance range of 5%. When comparing the experimental and control groups, the variables length, weight, and Lee's index presented no significant differences. In the x-ray analysis, at 8 weeks, the Co-L1 and Co-Mn measurements were reduced, whereas the Ba-So increased, with significant differences. At 16 weeks, the L1-O, Po-Ba, and E-Mu measurements decreased; however, Co-Gn registered a greater value with significant differences. The alkaline phosphatase levels fell significantly at week 16 in the experimental group. In conclusion, the reduction of permanent nasal respiratory flow is related to modifications in facial growth at 8 and 16 weeks and to the reduction of alkaline phosphatases at 16 weeks.

La respiración se considera una función vital, dependiente de factores como la permeabilidad adecuada de la vía nasal, vinculada con funciones fisiológicas, procesos intelectuales y crecimiento cráneofacial. El objetivo de este estudio fue determinar los cambios en el crecimiento y desarrollo óseo cráneo maxilo facial de ratas Sprague Dawley sometidas a obstrucción nasal unilateral experimental permanente. Se utilizaron 24 ratas macho neonatas, randomizadas y divididas en grupo control y experimental. Fue realizada obstrucción nasal de la narina derecha y realizadas mediciones de peso, longitud e índice de Lee a las 8 y 16 semanas. Se efectuaron radiografías cráneomandibulares a cada animal, obteniendo medidas lineales de neuro y viscerocráneo. Posteriormente se realizó análisis bioquímico, para medir la concentración de fosfatasa alcalina. Los resultados fueron analizados en el software SPSS, realizándose análisis descriptivo, empleando prueba T para muestras independientes comparando características basales, cefalométricas y bioquímicas entre los grupos control y experimental, considerando un umbral de significancia de 5 %. Al comparar los grupos control y experimental, las variables longitud, peso e índice de Lee no presentaron diferencias significativas. En el análisis radiográfico, a las 8 semanas, las medidas Co-L1 y Co-Mn presentaron reducción, mientras que Ba-So aumentó, con diferencias significativas. A las 16 semanas, las medidas L1-O, Po-Ba y E-Mu disminuyeron, sin embargo, Co-Gn registró un mayor valor, con diferencias significativas. Los niveles de fosfatasa alcalina disminuyeron significativamente en la semana 16 en el grupo experimental. En conclusión, la reducción de flujo respiratorio nasal permanente se relaciona con modificaciones del crecimiento facial a las 8 y 16 semanas y con la reducción de ALK en análisis a las 16 semanas.

The 27 Facial Sutures: Timing and Clinical Consequences of Closure.

SUMMARY: Facial sutures contribute significantly to postnatal facial development, but their potential role in craniofacial disease is understudied. Since interest in their development and physiology peaked in the mid-twentieth century, facial sutures have not garnered nearly the same clinical research interest as calvarial sutures or cranial base endochondral articulations. In addition to reinforcing the complex structure of the facial skeleton, facial sutures absorb mechanical stress and generally remain patent into and beyond adolescence, as they mediate growth and refine the shape of facial bones. However, premature closure of these sites of postnatal osteogenesis leads to disrupted growth vectors and consequent dysmorphologies. Although abnormality in individual sutures results in isolated facial deformities, we posit that generalized abnormality across multiple sutures may be involved in complex craniofacial conditions such as syndromic craniosynostosis. In this work, the authors comprehensively review 27 key facial sutures, including physiologic maturation and closure, contributions to postnatal facial development, and clinical consequences of premature closure.

High mobility group AT-hook 2 regulates osteoblast differentiation and facial bone development.

The mutation and deletion of high mobility group AT-hook 2 (Hmga2) gene exhibit skeletal malformation, but almost nothing is known about the mechanism. This study examined morphological anomaly of facial bone in Hmga2-/- mice and osteoblast differentiation of pre-osteoblast MC3T3-E1 cells with Hmga2 gene knockout (A2KO). Hmga2-/- mice showed the size reduction of anterior frontal part of facial bones. Hmga2 protein and mRNA were expressed in mesenchymal cells at ossification area of nasal bone. A2KO cells differentiation into osteoblasts after reaching the proliferation plateau was strongly suppressed by alizarin red and alkaline phosphatase staining analyses. Expression of osteoblast-related genes, especially Osterix, was down-regulated in A2KO cells. These results demonstrate a close association of Hmga2 with osteoblast differentiation of mesenchymal cells and bone growth. Although future studies are needed, the present study suggests an involvement of Hmga2 in osteoblast-genesis and bone growth.

Developmental changes of the facial skeleton from birth to 18 years within a South African cohort (A computed tomography study).

Skeletal remains are often found on a crime scene in which a forensic anthropologist is then consulted to create a biological profile, which includes the estimation of age, sex, ancestry and stature. The viscerocranium plays an important role in the formation of a biological profile. However, to utilise the viscerocranium for age estimation, population specific normative data and knowledge of the development of the viscerocranium is required. Therefore, this study aimed to investigate the developmental changes from birth to 18 years of age of the facial skeleton of individuals from a South African cohort. This study comprised of 239 computed tomography (CT) scans (128 males; 111 females). The viscerocranium was subdivided into five regions viz.: orbital, nasal, midfacial, maxillary and mandibular. The linear parameters in each region were correlated to age to identify the developmental growth patterns of the viscerocranial regions according to male and female. The measurements which displayed the highest correlations with age were used to develop formulas which could be used for age estimation. The results of this study showed that the measurements in the orbital, midfacial, maxillary and mandibular regions experienced rapid growth between 0 and 5 years of age, with the nasal region increasing steadily over time. It was noted that males displayed overall larger measurements than females except for the anterior interorbital distance and both right and left zygomatic arch lengths (ZAL). Although only the left orbital height, nasal aperture height and mandible width displayed statistically significant size differences according to sex (p ≤ 0.05). The measurements which showed the highest correlations to age were the zygomatic arch distance (r = 0.8842, p < 0.001), ZAL (right: r = 0.8929, p < 0.001; left: r = 0.8656, p < 0.001) and the mandible width (r = 0.8444, p < 0.001). Formulas were derived for the measurements that could be used to forensically estimate age within a subadult cohort.

Age estimation of immature human skeletal remains from mandibular and cranial bone dimensions in the postnatal period.

Age estimation is one of the crucial first steps in the identification of human skeletal remains in both forensic and archeological contexts. In the postnatal period, age is traditionally estimated from dental development or skeletal growth, typically long bone diaphyseal length. However, in many occasions other methods are required. This study provides alternative means of estimating age of juvenile remains from the size of several cranial bones and the mandible. A sample of 185 identified juvenile skeletons between birth and 13 years of age from two European collections were used (Lisbon and Spitalfields). Measurements of the frontal, occipital-lateralis, occipital-basilaris, occipital-squamous, zygomatic, maxilla, and mandible were used to calculate classical calibration regression formulae for the sexes combined. The sample was divided into three age groups birth-2 years, 2-6 years, and 2-12.9 years, depending on bone and its growth trajectory. For all the bones, measurements of the youngest age groups yielded the most precise age estimates. The vault bones on average yielded the best performing models, with the frontal bone having the most precise of all. The mandible performed on par with the best performing cranial bones, particularly in individuals under the age of 2 years. This study provides one of the most comprehensive approaches to juvenile age estimation based on bones of the skull, providing a resource that potentially can help estimate age of juvenile skeletons from a variety of circumstances.

Effects of 4-Hexylresorcinol on Craniofacial Growth in Rats.

4-Hexylresorcinol (4HR) has been used as a food additive, however, it has been recently demonstrated as a Class I histone deacetylase inhibitor (HDACi). Unlike other HDACi, 4HR can be taken through foods. Unfortunately, some HDACi have an influence on craniofacial growth, therefore, the purpose of this study was to evaluate the effects of 4HR on craniofacial growth. Saos-2 cells (osteoblast-like cells) were used for the evaluation of HDACi and its associated activities after 4HR administration. For the evaluation of craniofacial growth, 12.8 mg/kg of 4HR was administered weekly to 4 week old rats (male: 10, female: 10) for 12 weeks. Ten rats were used for untreated control (males: 5, females: 5). Body weight was recorded every week. Serum and head samples were collected at 12 weeks after initial administration. Craniofacial growth was evaluated by micro-computerized tomography. Serum was used for ELISA (testosterone and estrogen) and immunoprecipitation high-performance liquid chromatography (IP-HPLC). The administration of 4HR (1-100 µM) showed significant HDACi activity (p < 0.05). Body weight was significantly different in male rats (p < 0.05), and mandibular size was significantly smaller in 4HR-treated male rats with reduced testosterone levels. However, the mandibular size was significantly higher in 4HR treated female rats with increased growth hormone levels. In conclusion, 4HR had HDACi activity in Saos-2 cells. The administration of 4HR on growing rats showed different responses in body weight and mandibular size between sexes.

Quick septoplasty in children: Long-term effects on nasal breathing and dentofacial morphology. A prospective cephalometric study.

OBJECTIVE: Many authors have investigated the most appropriate surgical approach to the deviated septum in childhood, considering the obligate mouth-breathing habit a possible cause of malocclusion and disharmonious development of the facial skeleton in growing kids. Nevertheless, controversies still remain about the long-term functional/esthetic results of such procedures, mainly due to the duration of the follow-up and possible confounding factors. METHODS: 111 Caucasian children (age range: 6-13 years) were submitted to a personal "Quick" septoplasty surgical technique between 2005 and 2010. Preoperative otorhinolaryngological examination using flexible nasal endoscopy, anterior active rhinomanometry (AAR), Nasal Obstruction Septoplasty Effectiveness (NOSE) questionnaire, and dentofacial evaluation (including cephalometry) were performed. Postoperative AAR, NOSE questionnaire and cephalometric assessment were carried out in all patients at the age of 18 years. Informed consent was obtained from children's parents as appropriate. RESULTS: No surgical complication was recorded. Among our patients, a significant (p <0.001) improvement of nasal breathing resistances at AAR and NOSE questionnaire scores was found after surgery. A significant improvement in cephalometric/dental parameters (gonial angle values, anterior facial height, prevalence of class I occlusion, maxillary intermolar width, and cross-bite frequency) was noticed after the follow-up with respect to preoperative conditions. CONCLUSION: The "Quick" septoplasty technique described is a practical and conservative procedure with a low complication rate that offers long-term favourable results for the correction of nasal septum deviations in children. Nasal-breathing restoration may favor a physiological and harmonious development of craniofacial and dental structures in offspring.

A model of developmental canalization, applied to human cranial form.

Developmental mechanisms that canalize or compensate perturbations of organismal development (targeted or compensatory growth) are widely considered a prerequisite of individual health and the evolution of complex life, but little is known about the nature of these mechanisms. It is even unclear if and how a "target trajectory" of individual development is encoded in the organism's genetic-developmental system or, instead, emerges as an epiphenomenon. Here we develop a statistical model of developmental canalization based on an extended autoregressive model. We show that under certain assumptions the strength of canalization and the amount of canalized variance in a population can be estimated, or at least approximated, from longitudinal phenotypic measurements, even if the target trajectories are unobserved. We extend this model to multivariate measures and discuss reifications of the ensuing parameter matrix. We apply these approaches to longitudinal geometric morphometric data on human postnatal craniofacial size and shape as well as to the size of the frontal sinuses. Craniofacial size showed strong developmental canalization during the first 5 years of life, leading to a 50% reduction of cross-sectional size variance, followed by a continual increase in variance during puberty. Frontal sinus size, by contrast, did not show any signs of canalization. Total variance of craniofacial shape decreased slightly until about 5 years of age and increased thereafter. However, different features of craniofacial shape showed very different developmental dynamics. Whereas the relative dimensions of the nasopharynx showed strong canalization and a reduction of variance throughout postnatal development, facial orientation continually increased in variance. Some of the signals of canalization may owe to independent variation in developmental timing of cranial components, but our results indicate evolved, partly mechanically induced mechanisms of canalization that ensure properly sized upper airways and facial dimensions.

The Special Developmental Biology of Craniofacial Tissues Enables the Understanding of Oral and Maxillofacial Physiology and Diseases.

Maxillofacial hard tissues have several differences compared to bones of other localizations of the human body. These could be due to the different embryological development of the jaw bones compared to the extracranial skeleton. In particular, the immigration of neuroectodermally differentiated cells of the cranial neural crest (CNC) plays an important role. These cells differ from the mesenchymal structures of the extracranial skeleton. In the ontogenesis of the jaw bones, the development via the intermediate stage of the pharyngeal arches is another special developmental feature. The aim of this review was to illustrate how the development of maxillofacial hard tissues occurs via the cranial neural crest and pharyngeal arches, and what significance this could have for relevant pathologies in maxillofacial surgery, dentistry and orthodontic therapy. The pathogenesis of various growth anomalies and certain syndromes will also be discussed.

A developmental stage-specific network approach for studying dynamic co-regulation of transcription factors and microRNAs during craniofacial development.

Craniofacial development is regulated through dynamic and complex mechanisms that involve various signaling cascades and gene regulations. Disruption of such regulations can result in craniofacial birth defects. Here, we propose the first developmental stage-specific network approach by integrating two crucial regulators, transcription factors (TFs) and microRNAs (miRNAs), to study their co-regulation during craniofacial development. Specifically, we used TFs, miRNAs and non-TF genes to form feed-forward loops (FFLs) using genomic data covering mouse embryonic days E10.5 to E14.5. We identified key novel regulators (TFs Foxm1, Hif1a, Zbtb16, Myog, Myod1 and Tcf7, and miRNAs miR-340-5p and miR-129-5p) and target genes (Col1a1, Sgms2 and Slc8a3) expression of which changed in a developmental stage-dependent manner. We found that the Wnt-FoxO-Hippo pathway (from E10.5 to E11.5), tissue remodeling (from E12.5 to E13.5) and miR-129-5p-mediated Col1a1 regulation (from E10.5 to E14.5) might play crucial roles in craniofacial development. Enrichment analyses further suggested their functions. Our experiments validated the regulatory roles of miR-340-5p and Foxm1 in the Wnt-FoxO-Hippo subnetwork, as well as the role of miR-129-5p in the miR-129-5p-Col1a1 subnetwork. Thus, our study helps understand the comprehensive regulatory mechanisms for craniofacial development.

Differences among families in craniofacial shape at early life-stages of Arctic charr (Salvelinus alpinus).

BACKGROUND: Organismal fitness can be determined at early life-stages, but phenotypic variation at early life-stages is rarely considered in studies on evolutionary diversification. The trophic apparatus has been shown to contribute to sympatric resource-mediated divergence in several taxa. However, processes underlying diversification in trophic traits are poorly understood. Using phenotypically variable Icelandic Arctic charr (Salvelinus alpinus), we reared offspring from multiple families under standardized laboratory conditions and tested to what extent family (i.e. direct genetic and maternal effects) contributes to offspring morphology at hatching (H) and first feeding (FF). To understand the underlying mechanisms behind early life-stage variation in morphology, we examined how craniofacial shape varied according to family, offspring size, egg size and candidate gene expression. RESULTS: Craniofacial shape (i.e. the Meckel's cartilage and hyoid arch) was more variable between families than within families both across and within developmental stages. Differences in craniofacial morphology between developmental stages correlated with offspring size, whilst within developmental stages only shape at FF correlated with offspring size, as well as female mean egg size. Larger offspring and offspring from females with larger eggs consistently had a wider hyoid arch and contracted Meckel's cartilage in comparison to smaller offspring. CONCLUSIONS: This study provides evidence for family-level variation in early life-stage trophic morphology, indicating the potential for parental effects to facilitate resource polymorphism.

IFT20 is critical for collagen biosynthesis in craniofacial bone formation.

Intraflagellar transport (IFT) is essential for assembling primary cilia required for bone formation. Disruption of IFT frequently leads to bone defects in humans. While it has been well studied about the function of IFT in osteogenic cell proliferation and differentiation, little is known about its role in collagen biosynthesis during bone formation. Here we show that IFT20, the smallest IFT protein in the IFT-B complex, is important for collagen biosynthesis in mice. Deletion of Ift20 in craniofacial osteoblasts displayed bone defects in the face. While collagen protein levels are unaffected by loss of Ift20, collagen cross-linking was significantly altered. In both Ift20:Wnt1-Cre and Ift20:Ocn-Cre mice the bones exhibit increased hydroxylysine-aldehyde deived cross-linking, and decreased lysine-aldehyde derived cross-linking. To obtain insight into the molecular mechanisms, we examined the expression levels of telopeptidyl lysyl hydroxylase 2 (LH2), and associated chaperone complexes. The results demonstrated that, while LH2 levels were unaffected by loss of Ift20, its chaperone, FKBP65, was significantly increased in Ift20:Wnt1-Cre and Ift20:Ocn-Cre mouse calvaria as well as femurs. These results suggest that IFT20 plays a pivotal role in collagen biosynthesis by regulating, in part, telopeptidyl lysine hydroxylation and cross-linking in bone. To the best of our knowledge, this is the first to demonstrate that the IFT components control collagen post-translational modifications. This provides a novel insight into the craniofacial bone defects associated with craniofacial skeletal ciliopathies.

Estimating sex and age from a face: a forensic approach using machine learning based on photo-anthropometric indexes of the Brazilian population.

The facial analysis permits many investigations, some of the most important of which are craniofacial identification, facial recognition, and age and sex estimation. In forensics, photo-anthropometry describes the study of facial growth and allows the identification of patterns in facial skull development, for example, by using a group of cephalometric landmarks to estimate anthropological information. Previous works presented, as indirect applications, the use of photo-anthropometric measurements to estimate anthropological information such as age and sex. In several areas, automation of manual procedures has achieved advantages over and similar measurement confidence as a forensic expert. This manuscript presents an approach using photo-anthropometric indexes, generated from frontal faces cephalometric landmarks of the Brazilian population, to create an artificial neural network classifier that allows the estimation of anthropological information, in this specific case age and sex. This work is focused on four tasks: (i) sex estimation on ages from 5 to 22 years old, evaluating the interference of age on sex estimation; (ii) age estimation from photo-anthropometric indexes for four age intervals (1 year, 2 years, 4 years, and 5 years); (iii) age group estimation for thresholds of over 14 and over 18 years old; and; (iv) the provision of a new data set, available for academic purposes only, with a large and complete set of facial photo-anthropometric points marked and checked by forensic experts, measured from over 18,000 faces of individuals from Brazil over the last 4 years. The proposed binary classifier obtained significant results, using this new data set, for the sex estimation of individuals over 14 years old, achieving accuracy values higher than 0.85 by the F1 measure. For age estimation, the accuracy results are 0.72 for the F1 measure with an age interval of 5 years. For the age group estimation, the F1 measures of accuracy are higher than 0.93 and 0.83 for thresholds of 14 and 18 years, respectively.

Mutant COMP shapes growth and development of skull and facial structures in mice and humans.

BACKGROUND: Cartilage oligomeric matrix protein (COMP) is an important extracellular matrix protein primarily functioning in the musculoskeletal tissues and especially endochondral bone growth. Mutations in COMP cause the skeletal dysplasia pseudoachondroplasia (PSACH) that is characterized by short limbs and fingers, joint laxity, and abnormalities but a striking lack of skull and facial abnormalities. METHODS: This study examined both mice and humans to determine how mutant-COMP affects face and skull growth. RESULTS: Mutant COMP (MT-COMP) mice were phenotypically distinct. Snout length and skull height were diminished in MT-COMP mouse and the face more closely resembled younger controls. Three-dimensional facial measurements of PSACH faces showed widely spaced eyes, reduced lower facial height, and decreased nasal protrusion, which correlated with a more juvenile appearing face. Neither MT-COMP mice nor PSACH individuals show midface hypoplasia usually associated with abnormal endochondral bone growth. MT-COMP mice do show delayed endochondral and membranous skull ossification that normalizes with age. CONCLUSION: Therefore, mutant-COMP affects both endochondral and intramembranous bones of the skull resulting in a reduction of the nose and lower facial height in mice and humans, in addition to its well-defined role in the growth plate chondrocytes.

Relación entre los patrones de crecimiento facial y la maduración dental y esquelética en los pacientes en crecimiento / Relationship between facial growth patterns and dental and skeletal maturation in growing patients

El objetivo de este trabajo es realizar una revisión bibliográfica para evaluar la posible relación entre los patrones de crecimiento facial con la maduración dental y esquelética en pacientes en crecimiento. La maduración dental y esquelética son dos indicadores que nos permiten es-tablecer en qué momento del desarrollo se encuentra el paciente en crecimiento. Actualmente, para la valoración de la maduración esquelética se emplea la visualización de las vértebras cervicales en la radiografía lateral de cráneo, ya que es un método que permite disminuir la exposición radiológica del paciente que se encuentra en estudio ortodóncico, en comparación con la radiografía de muñe-ca, la cual suponía la realización de una radiografía adicional. Para el análisis de la maduración dental, la radiografía panorámica es la más empleada mediante la visualización del desarrollo de los siete dientes mandibulares izquierdos. Los pacientes que se encuentran en edad infantil presentan diversos patrones de crecimiento facial, entre los que podemos encontrar, braquifacial, mesofacial y dolicofacial. Para poder establecer el patrón de crecimiento que presenta el paciente existen diversos métodos que, a través de mediciones cefalométricas, nos clasifican al paciente en uno de los tres grupos. Existen diversos trabajos que estudian la posible relación entre los patrones de crecimiento facial y la maduración dental y esquelética. Según la literatura revisada podemos concluir que los pacientes que presentan un patrón de crecimiento vertical muestran una maduración dental y esquelética más avanzada en comparación con los pacientes que presentan un patrón de crecimiento horizontal

The objective of this paper is to present a bibliographical review to evaluate the possible relationship between facial growth patterns and dental and skeletal maturation in growing patients.Dental and skeletal maturation are two indicators that allow us to establish at what point the patient is in his growth. Currently, for the evaluation of skeletal maturation, the visualisation of the cervical vertebrae in the lateral x-ray of the skull is used, since this is a method that diminishes the radiological exposure of the patient undergoing an orthodontic study, in comparison with the x-ray of the wrist, which means taking an additional x-ray.For the analysis of dental maturation, the panoramic x-ray is the most used through visualisation of the development of the seven left jaw teeth. Children in young ages present different facial growth patterns, among which we can find brachyfacial, mesofacial and dolichofacial. In order to establish the growth pattern the patient presents, there are different methods that, through cephalometric measurements, classify the patient in one of the three groups. Different papers study the possible relationship between facial growth patterns and dental and skeletal maturation. According to the reviewed literature, we can conclude that the patients who present a vertical growth pattern show more advanced dental and skeletal maturation in comparison with the patients who present a horizontal growth pattern

Autophagy Regulates Craniofacial Bone Acquisition.

Increasing evidence has demonstrated the important role of autophagy in skeletal homeostasis; however, the role of autophagy in craniofacial bone development and acquisition is largely unknown. In this study, we investigated the effect of autophagy suppression on craniofacial bone acquisition by deleting Fip200 or Atg5, two essential autophagy genes, using Osterix-Cre (Osx-Cre). We found that the Osx-Cre transgene mildly decreased the bone mass of parietal bone but not frontal bone, and did not affect cranial base bone mass in adult mice. In the cranial vault, Fip200 or Atg5 deletion similarly decreased 50% bone mass of neural crest-derived frontal bone; Atg5 deletion decreased 50% and Fip200 deletion decreased 30% bone mass of mesoderm-derived parietal bone. In the cranial base, Fip200 or Atg5 deletion similarly decreased 30% bone mass of neural crest-derived presphenoid bone; Atg5 deletion decreased 30% and Fip200 deletion decreased 16% bone mass of mesoderm-derive basioccipital bone. Lastly, we used doxycycline treatment to inhibit the Osx-Cre expression until 2 months of age and showed that postnatal Fip200 deletion led to cranial vault bone mass decrease in association with a small increase in both bone volume/tissue volume and tissue mineral density. Altogether, this study demonstrated the important role of autophagy in craniofacial bone acquisition during development and postnatal growth.

Dental development and craniofacial morphology in school-age children.

INTRODUCTION: The growth of the craniofacial complex is important for establishing a balanced relationship among the teeth, jaws, and other facial structures. However, there is still a lack of information about craniofacial parameters that are affected by the rate of dental development. The aim of this study was to investigate the association between dental development and craniofacial morphology in school-age children. METHODS: This study was embedded in the Generation R Study, Rotterdam, The Netherlands. In 3,896 children aged 8 to 11 years, dental development was assessed from panoramic radiographs and craniofacial morphology was assessed by combining cephalometric parameters into 9 uncorrelated principal components, each representing a distinct skeletal or dental craniofacial pattern. The statistical analysis was performed using linear and nonlinear regression model. RESULTS: Dental development was positively associated with the bimaxillary growth (ß = 0.04; 95% CI 0.01 to 0.08). Children with above-average dental development had a tendency toward Class II jaw relationship (ß = -0.08; 95% CI -0.13 to -0.04). Regarding dental parameters, the proclination increased for incisors and lips with advanced dental development (ß = 0.15 [95% CI 0.10 to 0.19] and ß = 0.13 [95% CI 0.09 to 0.17], respectively), but the incisor proclination remained more pronounced in children that had above-average dental development. CONCLUSIONS: The findings of this large population-based study show that dental development is associated with specific dental and skeletal cephalometric characteristics in school-age children. Further longitudinal studies are necessary to confirm the observed effects over time.

Stem cells of the suture mesenchyme in craniofacial bone development, repair and regeneration.

Development of the skeleton is mediated through two distinct ossification mechanisms. Craniofacial bones are formed mainly through intramembranous ossification, a mechanism different from endochondral ossification required for development of the body skeleton. The skeletal structures are quite distinct between the two, thus they are likely to have their unique stem cell populations. The sutures serve as the growth center critical for healthy development of the craniofacial skeleton. Defects in suture morphogenesis cause its premature closure, resulting in development of craniosynostosis, a devastating disease affecting 1 in ~2,500 individuals. The suture mesenchyme has been postulated to act as the niche of skeletal stem cells essential for calvarial morphogenesis. However, very limited knowledge is available for suture biology and suture stem cells (SuSCs) have yet to be isolated. Here we report the first evidence for identification and isolation of a stem cell population residing in the suture midline. Genetic labeling of SuSCs shows their ability to self-renew and continually give rise to mature cell types over a 1-year monitoring period. They maintain their localization in the niches constantly produce skeletogenic descendants during calvarial development and homeostastic maintenance. Upon injury, SuSCs expand drastically surrounding the skeletogenic mesenchyme, migrate to the damaged site and contribute directly to skeletal repair in a cell autonomous fashion. The regeneration, pluripotency and frequency of SuSCs are also determined using limiting dilution transplantation. In vivo clonal expansion analysis demonstrates a single SuSC capable of generating bones. Furthermore, SuSC transplantation into injured calvaria facilitates the healing processes through direct engraftments. Our findings demonstrate SuSCs are bona fide skeletal stem cells ideally suited for cell-based craniofacial bone therapy as they possess abilities to engraft, differentiate.(Presented at the 1980th Meeting, April 16, 2019).

p63 establishes epithelial enhancers at critical craniofacial development genes.

The transcription factor p63 is a key mediator of epidermal development. Point mutations in p63 in patients lead to developmental defects, including orofacial clefting. To date, knowledge on how pivotal the role of p63 is in human craniofacial development is limited. Using an inducible transdifferentiation model, combined with epigenomic sequencing and multicohort meta-analysis of genome-wide association studies data, we show that p63 establishes enhancers at craniofacial development genes to modulate their transcription. Disease-specific substitution mutation in the DNA binding domain or sterile alpha motif protein interaction domain of p63, respectively, eliminates or reduces establishment of these enhancers. We show that enhancers established by p63 are highly enriched for single-nucleotide polymorphisms associated with nonsyndromic cleft lip ± cleft palate (CL/P). These orthogonal approaches indicate a strong molecular link between p63 enhancer function and CL/P, illuminating molecular mechanisms underlying this developmental defect and revealing vital regulatory elements and new candidate causative genes.

Variation in facial bone growth remodeling in prehistoric populations from southern South America.

OBJECTIVES: To assess the intraspecific variation in bone remodeling patterns in modern humans, we studied two populations from southern South America that represent the extremes of morphological variation in this region. We particularly analyzed the ontogenetic changes in the patterns of bone growth remodeling and compared the patterns between samples. MATERIALS AND METHODS: We obtained high-resolution casts of the periosteal surface of the upper and middle face of subadults (n = 36) and adult (n = 36) individuals from a sample of hunter-gatherers from Patagonia and a sample of horticulturists from Northwest Argentina. The areas of bone formation and resorption were registered using an incident-light microscope. We then estimated the average bone remodeling map by sample and age, and performed principal component analysis and multivariate regressions to assess the extension and distribution of these areas across ontogeny and between samples. RESULTS: We found that the remodeling pattern of the glabella, supraorbital arch, frontal process of the maxilla, and a large part of the zygomatic bone is relatively constant in subadults and adults from both sample with a clear predominance of bone formation. In contrast, the middle face is characterized by the spatial alternation between formation and resorption areas, and greater variation with age and between samples. The main differences were found in areas related to chewing and muscle insertions. CONCLUSIONS: Our study provides the first evidence of interpopulation variation in bone growth remodeling and suggests that biomechanical factors can influence the observed patterns. It also underlines the need to account for ecological factors in within and between species comparisons.