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Aim and Objective: The study aimed to explore the correlation between dermatoglyphic patterns and quantitative palatal anatomic variables in individuals with different growth patterns., Materials and Method: A cross-sectional study was conducted involving 126 healthy patients aged 17-25 years. Participants were divided into three groups based on growth patterns: average, vertical, and horizontal. Dermatoglyphic patterns were recorded using an optical fingerprint sensor, and palatal characteristics were measured using digital software. Palatal characteristics, including intercanine width, intermolar width, and palatal depth, were measured using digital software. The results were statistically analyzed., Results: Significant differences were observed in ridge counts among the three growth patterns. The average growth pattern showed lower ridge counts compared to the vertical and horizontal growth patterns. Dermatoglyphic patterns, such as double loops and tented arches, were significantly higher in the horizontal growth pattern. Weak correlations were found between certain dermatoglyphic patterns and palatal characteristics, with simple arch patterns showing a negative correlation with inter-canine width and symmetrical whorl patterns showing a positive correlation with palatal depth. Loop patterns, spiral patterns, double loop patterns, symmetrical whorl, and simple arch patterns were significant predictors of growth patterns., Conclusion: This study revealed distinct dermatoglyphic patterns and ridge counts among individuals with different growth patterns. Weak correlations were observed between dermatoglyphic patterns and palatal characteristics. However, the predictive value of dermatoglyphics for skeletal malocclusion requires further investigation. Understanding the relationships between dermatoglyphic patterns and craniofacial growth can provide valuable insights into genetic and developmental factors affecting dental and orthodontic conditions., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Introduction: This study aimed to use a finite element method to assess the stress on the miniscrews, skeletal and dental changes resulting from maxillary expansion using either a tooth-bone-borne (TBB) or a bone-borne (BB) device on patients with various skeletal maturation., Methods: Two types of expanders were modeled using SolidWorks. The design of the 2 expanders was similar, with the exception that the BB appliance lacked support for teeth (rods and bands). Both were placed on the palatal bones with the help of 4 miniscrews as bony anchorage. Five skeletal maturation stages were examined using suture maturation classification by Angelieri. A lateral displacement of 0.2 mm was applied to simulate 1 turn of jackscrew per day. The dental and skeletal changes from the treatment were quantified., Results: Stage A sutural maturation exhibited the greatest skeletal movement (0.13 mm/d) with a V-shaped displacement pattern. Stages B and C exhibited a more parallel expansion pattern. Stages D and E exhibited the least amount of skeletal movement with either device. Compared with the BB device, TBB demonstrated greater dental displacement. On average, the BB device exhibited a skeletal-to-dental expansion ratio of 70% from stage A to C compared with 49% with the TBB device., Conclusions: BB device showed more skeletal displacement than TBB, and the latter showed more dental side effects, particularly during stages B and C. The miniscrew-assisted rapid palatal expansion appliance was effective with sutural maturation stages A to C using the classification by Angelieri with a more parallel separation of the midpalatal sutures in stages B and C., (Copyright © 2024 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Failure of palatogenesis results in cleft palate, one of the most common congenital disabilities in humans. During the final phases of palatogenesis, the protective function of the peridermal cell layer must be eliminated for the medial edge epithelia to adhere properly, which is a prerequisite for the successful fusion of the secondary palate. However, a deeper understanding of the role and fate of the periderm in palatal adherence and fusion has been hampered due to a lack of appropriate periderm-specific genetic tools to examine this cell type in vivo. Here we used the cytokeratin-6A (Krt-6a) locus to develop both constitutive ( Krt6ai-Cre ) and inducible ( Krt6ai-Cre ERT2 ) periderm-specific Cre driver mouse lines. These novel lines allowed us to achieve both the spatial and temporal control needed to dissect the periderm fate on a cellular resolution during palatogenesis. Our studies suggest that, already before the opposing palatal shelves contact each other, at least some palatal periderm cells start to gradually lose their squamous periderm-like phenotype and dedifferentiate into cuboidal cells, reminiscent of the basal epithelial cells seen in the palatal midline seam. Moreover, we show that transforming growth factor-β (TGF-β) signaling plays a critical periderm-specific role in palatogenesis. Thirty-three percent of embryos lacking a gene encoding the TGF-β type I receptor ( Tgfbr1 ) in the periderm display a complete cleft of the secondary palate. Our subsequent experiments demonstrated that Tgfbr1- deficient periderm fails to undergo appropriate dedifferentiation. These studies define the periderm cell fate during palatogenesis and reveal a novel, critical role for TGF-β signaling in periderm dedifferentiation, which is a prerequisite for appropriate palatal epithelial adhesion and fusion.

Palatogenesis is affected by many factors, including gene polymorphisms and exposure to toxic chemicals during sensitive developmental periods. Cleft palate is one of the most common congenital anomalies, and ongoing efforts to elucidate the molecular mechanisms underlying palatogenesis are providing useful insights to reduce the risk of this disorder. To identify novel potential regulators of palatogenesis, we analyzed public transcriptome datasets from a mouse model of cleft palate caused by selective deletion of transforming growth factor-β (TGFβ) receptor type 2 in cranial neural crest cells. We identified the homeobox transcription factor Mohawk (Mkx) as a gene downregulated in the maxilla of TGFβ knockout mice compared with wild-type mice. To examine the role of mkx in palatogenesis, we used CRISPR/Cas9 editing to generate zebrafish with impaired expression of mkxa and mkxb, the zebrafish homologs of Mkx. We found that mkx crispants expressed reduced levels of gli1, a critical transcription factor in the Sonic hedgehog (SHH) signaling pathway that plays an important role in the regulation of palatogenesis. Furthermore, we found that mkxa -/- zebrafish were more susceptible than mkxa +/+ zebrafish to the deleterious effects of cyclopamine, an inhibitor of SHH signaling, on upper jaw development. These results suggest that Mkx may be involved in palatogenesis regulated by TGFβ and SHH signaling, and that impairment in Mkx function may be related to the etiology of cleft palate., (© 2021 Japanese Teratology Society.)

Introduction: The objective of this study was to classify median palatine suture (MPS) maturation type in young and adult patients. Additionally, we compared MPS maturity type and density based on sex and growth status., Materials and Methods: In this retrospective cone beam computed tomography study, we included a total of 221 subjects, grouped based on sex and growth status. Once scans were aligned and oriented in the sagittal view, we conducted our evaluations on the axial sections. Based on interdigitation and shape, the MPS were categorized into Maturation Types A through E. Additionally, MPS density was measured as Hounsfield unit equivalent pixel intensity value scale for anterior and posterior sutural regions., Results: The majority of male (39%) and female (42%) subjects had MPS Maturation Type C. A maximum number of growing (42%) patients had Type C and nongrowing subjects (39%) had Type E sutures. The sex comparison showed significantly lower (p < .001) MPS density for both anterior and posterior regions in males when compared to females. Additionally, for the posterior region, nongrowing males had significantly lower (p < .001) MPS density when compared to nongrowing females. Subgroup comparisons of the MPS densities between growing and nongrowing males and growing and nongrowing females showed a significant difference (p < .001)., Conclusion: Classification of the MPS based on the maturation types provides a reliable predictor for orthodontic treatment planning. MPS density is significantly higher in females as compared to males. Similarly, nongrowing individuals have significantly higher MPS density compared to growing individuals for both anterior and posterior locations., (© 2020 Wiley Periodicals, LLC.)

Objective: To determine the ossification level of the midpalatal suture (MPS) in children, adolescents and adults from a Peruvian sample; according to the method proposed by Angelieri et al. with cone-beam computed tomographies (CBCTs)., Material and Methods: The sample consisted of 315 CBCTs of 168 females and 147 males. The total sample was divided into 3 groups according to age and sex: children (n=77), adolescents (n=113) and adults (n=125). The images were mainly assessed in the axial plane using the Real Scan 2.0 software. The different periods of ossification of the MPS at the level of the vertical half of the palate was defined according to the five stages of Angieleri method (from A to E). The Student t-Test, Chi-square test, Kruskal-Wallis test and Spearman's Rho test were applied., Results: Chi 2 test results showed that the stages of MPS ossification depended on the age of the patient by age ranges (P<0.005). Multiple comparison tests affirmed that male and female subjects in the children group had fewer ossification stages than the adolescent and adult groups (P<0.001). Meanwhile, there were no statistically significant differences between the two older age groups. Finally, there was a moderate positive correlation between the stages of ossification of the MPS and the age in the male group (Rho=0.511)., Conclusions: The onset of MPS ossification was significantly related to subjects up to 12 years of age and more frequently in stages B and C in both sexes. There was no difference in MPS ossification in adolescents and adult subjects., (Copyright © 2021 CEO. Published by Elsevier Masson SAS. All rights reserved.)

The purpose of this study is to evaluate the changes in the palatal alveolar bone thickness and find the factors related to the resorption of the palatal alveolar bone caused by tooth movement after the maxillary incisors were retracted and intruded during orthodontic treatment. The study group comprised of 33 skeletal Class II malocclusion patients who underwent extraction for orthodontic treatment. Palatal alveolar bone thickness changes and resorption factors were identified and analyzed. The changes of maxillary central incisors and palatal alveolar bone thickness were measured, and the corresponding sample t test was performed using SPSS (IBM SPSS version 22). The amount of palatal alveolar bone resorption was measured and various parameters were analyzed to determine which factors affected it. Correlation analysis adopting the amount of palatal alveolar bone resorption as a dependent variable demonstrated that the SNB, mandibular plane angle, and the inclination of the maxillary central incisor were significantly correlated with before treatment. On the other hand, mandibular plane angle, angle of convexity, the inclination of the upper incisor, and the occlusal plane (UOP, POP) were significantly correlated with post-treatment. In addition, the variables related to palatal contour (PP to PAS, SN to PAS, palatal surface angle) and occlusal planes (UOP/POP) were significantly correlated with the difference in palatal bone resorption. During initial diagnosis, high angle class II with normal upper incisor inclination can be signs of high-risk factors. In addition, maintaining the occlusal plane during treatment helps to prevent palatal bone loss.

Aim: The aim of the current study was to evaluate the correlation of skeletal age based on cervical vertebrae maturation (CVM) stage and mid-palatal suture (MPS) maturation in an Iranian population., Material and Methods: This was a cross-sectional analytic study. A total number of 93 samples were included. Samples were taken from patients who were in CS3 to CS6 stages of CVM who had cone-beam computed tomography and lateral cephalometry based on inclusion criteria. The maturation of MPS was assessed based on the cone-beam computed tomography images. In the classification of maturation of MPS, there are five stages (A-E) and the suture fusion occurs in stage D. In stage E, the suture is fused completely. The CVM stage (CS1-6) was also assessed based on the lateral cephalograms. Data were analyzed using Spearman correlation with a significance level of 0.05., Results: A total of 51 female individuals with a mean age of 14.98 ± 4.806 and 42 male individuals with a mean age of 15.79 ± 5.135 participated in this study. The correlation coefficient between the CVM stage and MPS maturation was 0.691 in female and 0.754 in male individuals (P < 0.001). Stage D was correlated with CS4., Conclusion: The results demonstrated that CVM stages had a significant but moderate positive correlation with the maturation of MPS. Until CS3, the MPS has not been fused and in CS6 the MPS is fused definitely., (Copyright © 2020 World Federation of Orthodontists. Published by Elsevier Inc. All rights reserved.)

The morphogenesis of the mammalian secondary plate is a series of highly dynamic developmental process, including the palate shelves vertical outgrowth, elevation to the horizontal plane and complete fusion in the midline. Extracellular matrix (ECM) proteins not only form the basic infrastructure for palatal mesenchymal cells to adhere via integrins but also interact with cells to regulate their functions such as proliferation and differentiation. ECM remodeling is essential for palatal outgrowth, expansion, elevation, and fusion. Multiple signaling pathways important for palatogenesis such as FGF, TGF β, BMP, and SHH remodels ECM dynamics. Dysregulation of ECM such as HA synthesis or ECM breakdown enzymes MMPs or ADAMTS causes cleft palate in mouse models. A better understanding of ECM remodeling will contribute to revealing the pathogenesis of cleft palate.

Background: The molecular mechanisms of abnormal palatogenesis were investigated in this study. A key regulator, miR-106a-5p, and its target pathway were analyzed., Objectives: This research is trying to clarify the underlying mechanism of the modulation of miRNA transcription during the formation of cleft palate by 7T and 9.4T NMR metabolomic platforms., Method: Differentially expressed miRNAs and mRNAs were analyzed by microarray analysis and verified by qRT-PCR. The protein expression in TGFβ signaling pathways were analyzed by Western Blotting. The relationship between miR-106a-5p and TGFβ were analyzed by luciferase reporter assay. Cell apoptosis were analyzed by flow cytometer. And finally, the metabonomics were analyzed by NMR and multivariate data analysis models (MVDA)., Results: The expression of miR-106a-5p increased in cleft palatal tissue and negatively correlated with the protein level of Tgfbr2. The luciferase assay further proved that the tgfbr2 was a direct target of miR-106a-5p. In another aspect, miR-106a-5p increased apoptosis level in palatal mesenchymal cells, possibly because its inhibition of TGFβ signaling pathway. Moreover, low cholesterol and choline levels with high citric acid and lipid levels were observed by 7T and 9.4T NMR metabonomic analysis, which inferred the disorder of cell membrane synthesis in cleft palate formation. Furthermore, transformation from choline to phosphatidylcholine regulated by miR-106a-5p was also disrupted, resulting in phosphatidic choline synthesis disorder and reduced cell membrane synthesis., Conclusions: The regulatory mechanism of cleft palate was studied at transcriptional and metabolomics levels, which may provide important information in understanding the primary cause of this abnormality., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Cone beam computed tomography (CBCT) imaging techniques are the recent rage in the field of oral diagnostic imaging modality. It is noninvasive, faster and lacks anatomic superimposition. Earlier maxillary occlusal radiographs were used to assess and evaluate the mid palatal suture, but being a two dimensional imaging modality it could not assess the ossification process which takes place in multiple planes mostly due to curved nature of the palate. In this study we assessed the mid palatal suture morphology and classify them according to the variants using CBCT images., Materials and Methods: A total of 200 CBCT scans (95 males and 105 females) were evaluated in the present study from the archives of an imaging center. As per Angelieri classification the midpalatal suture was classified into five categories (A-E) depending on the degree of ossification that had taken place. Statistical analysis was done by Chi Square test using SPSS version 23.0., Results: There is statistically significant difference present in the stages of maturity of mid palatal suture in various age groups with Stage B is most common in Group 1 (50%), Stage C most common in Group 2 (60%) and Group 3 (40%) and Stage E more common in Group 4 (50%)., Conclusion: The results of the present study showed a wide variation in the initiation time and the degree of ossification and morphology of the midpalatal suture in different age groups. Although there was an increase in the closure of the suture with aging, age is not a reliable criterion for determining the open or closed nature of the suture. This finding is important in providing an idea as to how diverse is the ossification of maxillary sutures.

Periodic patterning of iterative structures is a fundamental process during embryonic development, since these structures are diverse across the animal kingdom. Therefore, elucidating the molecular mechanisms in the formation of these structures promotes understanding of the process of organogenesis. Periodically patterned ridges, palatal rugae (situated on the hard palate of mammals), are an excellent experimental model to clarify the molecular mechanisms involved in the formation of periodic patterning of iterative structures. Primary cilia are involved in many biological events, including the regulation of signaling pathways such as Shh and non-canonical Wnt signaling. However, the role of primary cilia in the development of palatal rugae remains unclear. We found that primary cilia were localized to the oral cavity side of the interplacode epithelium of the palatal rugae, whereas restricted localization of primary cilia could not be detected in other regions. Next, we generated mice with a placodal conditional deletion of the primary cilia protein Ift88, using ShhCre mice (Ift88  fl/fl ;ShhCre). Highly disorganized palatal rugae were observed in Ift88  fl/fl ;ShhCre mice. Furthermore, by comparative in situ hybridization analysis, many Shh and non-canonical Wnt signaling-related molecules showed spatiotemporal expression patterns during palatal rugae development, including restricted expression in the epithelium (placodes and interplacodes) and mesenchyme. Some of these expression were found to be altered in Ift88  fl/fl ;ShhCre mice. Primary cilia is thus involved in development of palatal rugae., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to develop an accurate and intuitive semi-automatic segmentation technique to calculate an average maxillary arch and palatal growth profile for healthy newborns in their first year of life., Materials and Methods: Seventy babies born between 1985 and 1988 were included in this study. Each child had five impressions made in the first year after birth that were digitalized. A semi-automatic segmentation tool was developed and used to assess the maxillary dimensions. Finally, random effect models were built to describe the growth and build a simulation population of 10,000 newborns. The segmentation was tested for inter- and intra-observer variability., Results: The Pearson correlation coefficient for each of the variables was between 0.94 and 1.00, indicating high inter-observer agreement. The paired sample t test showed that, except for the tuberosity distance, there were small, but significant differences in the landmark placements between observers. Intra-observer repeatability was high, with Pearson correlation coefficients ranging from 0.87 to 1.00 for all measurements, and the mean differences were not significant. A third or second degree growth curve could be successfully made for each parameter., Conclusions: These findings indicated this method could be used for objective clinical evaluation of maxillary growth., Clinical Relevance: The resulting growth models can be used for growth studies in healthy newborns and for growth and treatment outcome studies in children with cleft lip and palate or other craniofacial anomalies.

Normal mammalian palatogenesis is a complex process that requires the occurrence of a tightly regulated series of specific and sequentially regulated cellular events. Cleft lip/palate (CLP), the most frequent craniofacial malformation birth defects, may occur if any of these events undergo abnormal interference. Such defects not only affect the patients, but also pose a financial risk for the families. In our recent study, the miniature pig was shown to be a valuable alternative large animal model for exploring human palate development by histology. However, few reports exist in the literature to document gene expression and function during swine palatogenesis. To better understand the genetic regulation of palate development, an mRNA expression profiling analysis was performed on miniature pigs, Sus scrofa . Five key developmental stages of miniature pigs from embryonic days (E) 30-50 were selected for transcriptome sequencing. Gene expression profiles in different palate development stages of miniature pigs were identified. Nine hundred twenty significant differentially expressed genes were identified, and the functional characteristics of these genes were determined by gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Some of these genes were associated with HH (hedgehog), WNT (wingless-type mouse mammary tumor virus integration site family), and MAPK (mitogen-activated protein kinase) signaling, etc., which were shown in the literature to affect palate development, while some genes, such as HIP (hedgehog interacting protein), WNT16 , MAPK10 , and LAMC2 (laminin subunit gamma 2), were additions to the current understanding of palate development. The present study provided a comprehensive analysis for understanding the dynamic gene regulation during palate development and provided potential ideas and resources to further study normal palate development and the etiology of cleft palate.

Cleft palate is a common oral and craniomaxillofacial birth defect. As the ideal surgery time is shortly after birth, clinical treatments should result in minimal disruption of theskeleton to allow tissue growth in children. A tissue-engineered graft was created in this study for cleft palate repair by integrating poly(1,8-octamethylene-citrate) (POC) with a decellularized amnion membrane (DAM-POC) to incorporate the advantages of both the synthetic polymer and the native tissue. The success of POC incorporation was confirmed by laser-induced breakdown spectroscopy and fluorescence detection. The DAM-POC scaffold showed a certain level of structure collapse and lower stiffness but better resistance to enzyme digestion than the native amnion and DAM scaffold. The DAM-POC scaffold is cell compatible when seeded with mesenchymal stem cells, as evidenced by adequate cell viability and improved alkaline phosphatase (ALP) activity and calcium deposit. A large palate defect was first surgically created in a young rat model and then repaired with the DAM-POC scaffold. Eight weeks postsurgery, histological study and CT scans showed nearly complete healing of both soft and hard tissues. In conclusion, we developed a cell-free, resorbable graft by incorporating and integrating a synthetic polymer with a human DAM. When the DAM-POC scaffold was applied to repair a large palate defect in young rats, it showed adequate biocompatibility as evidenced by its effectiveness in guiding hard and soft tissue regeneration and minimum interference with natural growth and palate development of rats. STATEMENT OF SIGNIFICANCE: Proper restoration of severe cleft palate remains a major challenge because of insufficient autologous soft tissues to close the open wounds, thereby causing high tension at the surgical junction, secondary palatal fistulas, wound contraction, scar tissue formation, and facial growth disturbances. In this study, we have developed a tissue-engineered graft through incorporating and integrating a synthetic polymer with the human amnion membrane for cleft palate repair. The significance of this study lies in our ability to develop a cell-free, resorbable graft that can provide a less surgically invasive option to cover the open defect and support palate regeneration and tissue growth. This technique could potentially advance soft and hard tissue regeneration in children with birth craniomaxillofacial defects., (Copyright © 2019 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Epithelial fusion underlies many vital organogenic processes during embryogenesis. Disruptions to these cause a significant number of human birth defects, including ocular coloboma. We provide robust spatial-temporal staging and unique anatomical detail of optic fissure closure (OFC) in the embryonic chick, including evidence for roles of apoptosis and epithelial remodelling. We performed complementary transcriptomic profiling and show that Netrin -1 ( NTN1 ) is precisely expressed in the chick fissure margin during fusion but is immediately downregulated after fusion. We further provide a combination of protein localisation and phenotypic evidence in chick, humans, mice and zebrafish that Netrin-1 has an evolutionarily conserved and essential requirement for OFC, and is likely to have an important role in palate fusion. Our data suggest that NTN1 is a strong candidate locus for human coloboma and other multi-system developmental fusion defects, and show that chick OFC is a powerful model for epithelial fusion research., Competing Interests: HH, JP, AP, SD, VT, HK, AY, LG, BB, JS, JR No competing interests declared, (© 2019, Hardy et al.)

Background: Marfan syndrome is a rare autosomal dominant inherited disease of the connective tissue associated with various craniofacial abnormalities. Aim of the present study was to assess the variability of palatal shape in a sample of 31 Marfan patients compared to a control group of no syndromic subjects, in two stages of dentition, by using 3D geometric morphometric analysis., Methods: Thirty one growing subjects with Marfan syndrome were selected and divided into two subgroups: MG1 with mixed dentition (10 M, 6F, mean age 7+/- 0.7 years), MG2 with permanent dentition (8 M, 7F, mean age 13+/- 0,5 years). Each subgroup was compared to a control group (CG1 mixed dentition, 9 M, 7F, mean age 7.6+/- 0.5 years; CG2 permanent dentition, 9 M, 6F, mean age 12.8+/- 0.7 years) matched on age, sex distribution, stage of dentition and skeletal maturation. Then the two subgroups were compared one to each other. For each patient maxillary dental casts were taken, scanned and digitized. 3D geometric morphometric methods were applied. Procrustes analysis was used and principal component analysis was performed to reveal the main patterns of palatal shape variation., Results: Both Marfan subgroups showed important reductions in the transversal plane associated with a deep palatal vault when compared to the control groups (MG1 vs CG1 P = 0,003; MG2 vs CG2 P = 0,07). Moreover a statistically significant difference between the palatal shape of MG1 and MG2 was found (P = 0.017) showing a significant worsening of palatal depth and constriction from mixed to permanent dentition in Marfan subjects., Conclusion: Marfan subjects showed a specific palatal morphology with maxillary constriction and deeper palatal vault when compared to a control group of healthy subjects. The constriction and the depth of the palatal vault in Marfan patients worsen from mixed dentition to permanent dentition more then in no syndromic subjects.

Objectives: To assess reliability and reproducibility of the individual assessment of midpalatal suture maturation in computed tomography among orthodontists and radiologists for potential diagnosis application., Materials and Methods: Sixty axial slices from cone-beam computed tomography and multi-slice CT scans of patients aged between 11 and 21 years old (33 females and 27 males) were selected. For the investigation of reliability and reproducibility of the method, two groups of examiners were established. The first group consisted of 11 orthodontists and the second consisted of 10 radiologists. Each group examined the images and performed individual assessment of the midpalatal suture maturation method twice within an interval of 21 days. During the first and second analyses, the sequence of images was randomized to reduce potential bias. Weighted Cohen's kappa was performed to assess inter- and intra-examiners' agreement. The percentage of perfect agreement and the number of stages apart for each disagreement were calculated. The significance level was P < .05., Results: The overall inter-examiner agreement was satisfactory in the first (kappa w : 0.37) and the second (kappa w : 0.34) analyses. Intra-examiner agreement outcomes were similar between orthodontists (kappa w : 0.44) and radiologists (kappa w : 0.41). The percentage of perfect agreement was 43.2%., Conclusions: The method for individual assessment of midpalatal suture maturation revealed potential reliability and reproducibility. However, the agreement rate observed in the present study was not high enough for a method designed for routine clinical applications.

Mammalian palate separates the oral and nasal cavities for normal feeding, breathing and speech. The palatal shelves are a pair of maxillary prominences that consist of the neural crest-derived mesenchyme and surrounding epithelium. Palatogenesis is completed by the fusion of the midline epithelial seam (MES) after the medial edge epithelium (MEE) cells make contact between the palatal shelves. Various cellular and molecular events, such as apoptosis, cell proliferation, cell migration, and epithelial-mesenchymal transition (EMT), are involved in palatogenesis. The Zeb family of transcription factors is an essential player during normal embryonic development. The distinct role of the Zeb family has not been thoroughly elucidated to date. In mouse palate, the Zeb family factors are expressed in the palatal mesenchyme until MEE contact. Interestingly, the expression of the Zeb family has also been observed in MES, which is already fused with the mesenchymal region. The regulatory roles of the Zeb family in palatogenesis have not been elucidated to date. The purpose of this study is to determine the Zeb family effects on the cellular events. To investigate the functions of the Zeb family, siRNA targeting Zeb family was used to treat in vitro organ culture for temporary inhibition of the Zeb family during palatogenesis. In the cultured palate containing siRNA, MES was clearly observed, and E-cadherin, an epithelial marker, was still expressed. Inhibition of the Zeb family results in the suppression of apoptosis, increased cell proliferation, and defective cell migration in the developing palate. Our data suggest that the Zeb family plays multiple roles in the stimulation and inhibition of apoptosis and cell proliferation and efficient mesenchymal cell migration during palatogenesis., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: To evaluate topographic and temporal aspects of premaxillary bone and premaxillary-maxillary suture, since they are fundamental anatomical elements little explored clinically., Methods: 1,138 human dry skulls were evaluated, of which 116 (10.19%) of the specimens were children, and 1,022 (89.81%) were adults. The skulls were photographed and the percentage of premaxillary-maxillary suture opening was determined. Subsequently the data were tabulated and submitted to statistical analysis, adopting a level of significance of 5%., Results: The progression of premaxillary suture closure from birth to 12 years of age was 3.72% per year. In 100% of the skulls up to 12 years, the premaxillary-maxillary suture open in the palatal region was observed, while 6.16% of adults presented different degrees of opening., Conclusions: The premaxilla exists in an independent way within the maxillary complex and the presence of the premaxilla-maxillary suture justifies the success of anteroposterior expansions to stimulate the growth of the middle third of the face, solving anatomical and functional problems.

Although minor salivary glands play a significant functional role in the oral cavity, their developmental morphology and cell differentiation has been scarcely studied. This study aimed to describe the development of rat palatine glands with regard to the ultrastructural morphology of the secretory cells and surrounding myoepithelial cells (MECs). Palatine glands from rats at embryonic ages (E) 18 and 20 days, and postnatal days (PN) 0, 3, 7, 10, 13, 21, 30, 42, and 60 were fixed and prepared for morphological analysis and immunocytochemical labeling of alpha-smooth muscle actin (α-SMA). At E18, epithelial cords were observed extending from the palatal epithelium and showed negative reactivity to α-SMA. After luminization at E20, the cells of immature acini accumulated secretory granules of various densities: electron-dense, electron-lucent and some empty-appearing granules. MECs were poorly differentiated at E20 and exhibited only slight α-SMA expression. At birth, mucous and serous cells were typically located around a common lumen. Thereafter, serous cells began to move to the periphery to form demilunes by PN7. The mucous secretory granules of intermediate electron density became predominant around PN13. At PN21, these granules were dramatically reduced in number and most of the acini in adults contained acinar cells with numerous electron-lucent granules, and a few serous demilune cells with electron-dense granules. After birth, MECs progressively accumulated actin microfilaments until prominent α-SMA expressing MECs invested the acini and the proximal part of the intercalated ducts in the adult. Anat Rec, 301:1820-1833, 2018. © 2018 Wiley Periodicals, Inc., (© 2018 Wiley Periodicals, Inc.)

Runx1 deficiency results in an anteriorly specific cleft palate at the boundary between the primary and secondary palates and in the first rugae area of the secondary palate in mice. However, the cellular and molecular pathogenesis underlying such regional specificity remain unknown. In this study, Runx1 epithelial-specific deletion led to the failed disintegration of the contacting palatal epithelium and markedly downregulated Tgfb3 expression in the primary palate and nasal septum. In culture, TGFB3 protein rescued the clefting of the mutant. Furthermore, Stat3 phosphorylation was disturbed in the corresponding cleft regions in Runx1 mutants. The Stat3 function was manifested by palatal fusion defects in culture following Stat3 inhibitor treatment with significant downregulation of Tgfb3. Tgfb3 is therefore a critical target of Runx1 signaling, and this signaling axis could be mediated by Stat3 activation. Interestingly, the expression of Socs3, an inhibitor of Stat3, was specific in the primary palate and upregulated by Runx1 deficiency. Thus, the involvement of Socs3 in Runx1-Tgfb3 signaling might explain, at least in part, the anteriorly specific downregulation of Tgfb3 expression and Stat3 activity in Runx1 mutants. This is the first study to show that the novel Runx1-Stat3-Tgfb3 axis is essential in anterior palatogenesis.

Background: Microdeletion of chromosome 22q11 is associated with significant developmental anomalies, including disruption of the cardiac outflow tract, thymic/parathyroid aplasia and cleft palate. Amongst the genes within this region, TBX1 is a major candidate for many of these developmental defects. Targeted deletion of Tbx1 in the mouse has provided significant insight into the function of this transcription factor during early development of the cardiac and pharyngeal systems. However, less is known about its role during palatogenesis. To assess the influence of Tbx1 function on gene expression profile within the developing palate we performed a microarray screen using total RNA isolated from the secondary palate of E13.5 mouse embryos wild type, heterozygous and mutant for Tbx1., Results: Expression-level filtering and statistical analysis revealed a total of 577 genes differentially expressed across genotypes. Data were clustered into 3 groups based on comparison between genotypes. Group A was composed of differentially expressed genes in mutant compared to wild type (n = 89); Group B included differentially expressed genes in heterozygous compared to wild type (n = 400) and Group C included differentially expressed genes in mutant compared to heterozygous (n = 88). High-throughput quantitative real-time PCR (RT-PCR) confirmed a total of 27 genes significantly changed between wild type and mutant; and 27 genes between heterozygote and mutant. Amongst these, the majority were present in both groups A and C (26 genes). Associations existed with hypertrophic cardiomyopathy, cardiac muscle contraction, dilated cardiomyopathy, focal adhesion, tight junction and calcium signalling pathways. No significant differences in gene expression were found between wild type and heterozygous palatal shelves., Conclusions: Significant differences in gene expression profile within the secondary palate of wild type and mutant embryos is consistent with a primary role for Tbx1 during palatogenesis.

Numerous studies have been conducted to understand the molecular mechanisms controlling mammalian secondary palate development such as growth, reorientation and fusion. However, little is known about the signaling factors regulating palate initiation. Mouse fibroblast growth factor (FGF) receptor 2 gene (Fgfr2) is expressed on E11.5 in the palate outgrowth within the maxillary process, in a region that is responsible for palate cell specification and shelf initiation. Fgfr2 continues to express in palate on E12.5 and E13.5 in both epithelial and mesenchymal cells, and inactivation of Fgfr2 expression in mesenchymal cells using floxed Fgfr2 allele and Osr2-Cre leads to cleft palate at various stages including reorientation, horizontal growth and fusion. Notably, some mutant embryos displayed no sign of palate shelf formation suggesting that FGF receptor 2 mediated FGF signaling may play an important role in palate initiation., (Copyright © 2018. Published by Elsevier Inc.)

The purpose of this study was to investigate the long-term effects of two-stage palatoplasty on the morphology of the maxillary alveolar arch and occlusion using plaster models of the maxilla and mandible obtained from patients with unilateral complete cleft lip and palate who also underwent orthodontic treatment. A total of 20 patients undergoing two-stage palatoplasty by Perko's method (Group T) were enrolled. Plaster models of the maxilla and mandible were obtained from each patient at Time 1, on commencement of orthodontic treatment in the mixed dentition period; at Time 2, on that of orthodontic treatment in the permanent dentition period; and at Time 3, on completion of active orthodontic treatment. Analysis of occlusion and morphological analysis were performed using a 3-dimensional measuring system. The results were compared with 15 patients who underwent one-stage palatoplasty by the push-back method using a mucoperiosteal flap (Group P). Alveolar morphology and the relationship between the maxilla and mandible were satisfactory in Group T. The palates in Group T were deeper and larger than those in Group P. Alveolar collapse in Group T was milder, and impairment of the alveolar morphology less notable than in Group P, as surgical invasion to the anterior alveolar region was avoided during the palatal growth period. These results suggest that two-stage palatoplasty is advantageous for jaw development.

This study describes the dentoalveolar and palatal growth during the first months of life. Knowledge concerning this development is essential to avoid unwanted events such as mucosal ulcerations or restriction of growth when cleft-lip and palate (CLP) patients are treated. The results involve the generation of CAD/CAM CLP-feeding plates. Intraoral impressions from 32 healthy newborns were taken monthly for 5 months, supplemented by measurements of body weight, length, and occipital-frontal head circumference. The casts were digitalized, and two observers manually selected defined anatomical landmarks on virtual 3-D models. The distances between these landmarks were evaluted. Statistical analysis included an inter-rater agreement analysis and the determination of growth. In total, 213 casts were analyzed, with 65 models excluded because of inaccuracies in impression-taking or cast production. Overall longitudinal growth was 20.3%, whereas transversal growth reached a maximum of 21.1%. Vertical growth was 32.4% at the tuberal level. On the basis of these results, a semiautomated series of feeding plates allowing for monthly expansion could be generated. The acquired data serve as a useful reference for other pediatric and orthofacial investigations and treatments. One such application is the automated, fully virtual manufacture of CLP-feeding plates based on only one impression-taking. Our data reveal when caution is needed to prevent ulceration. The series of plates generated can minimize the time-consuming impression-taking and the production of further plaster models. The method of measurement is suitable for documentary purposes. Clin. Anat. 30:846-854, 2017. © 2017 Wiley Periodicals, Inc., (© 2017 Wiley Periodicals, Inc.)

Normal cell cycle progression and proliferation of palatal mesenchymal cells are important for palatal development. As targets of miR-17-92, E2F transcription factors family has been suggested to induce the transcription of miR-17-92 in several cell types. In the present study, we sought to investigate whether this negative feedback loop exists in mouse PMCs and what the function of this negative feedback loop would be in palatal mesenchymal cells. Using GeneMANIA, we revealed that the most important function of experimentally verified targets of miR-17-92 is cell cycle regulation. E2F1 and E2F3, but not E2F2, were extensively expressed in mouse palate. Over-expression of E2F1 significantly increased the expression of all the members of miR-17-92. After increased by E2F1, miR-17 and miR-20a may negatively target E2F1, and thereby prevent the cells from excessive proliferation. We suggest that the negative feedback loop between E2F1 and miR-17-92 may contribute to palatal development by regulating the proliferation and cell cycle of palatal mesenchymal cells.

Introduction: We used cone-beam computed tomography to evaluate the maturation stages of the midpalatal sutures in children aged 11 to 15 years old. Maxillary expansion is successful for most patients in this age group, so we sought to identify the status of suture maturation in these subjects to use as a comparison for the prognosis of rapid maxillary expansion in older patients., Methods: Tomographic images in axial sections of the midpalatal sutures from 84 children (40 boys, 44 girls; ages, 11-15 years) were classified using a scale denoting the maturation stage of the midpalatal suture (A, B, C, D, and E). The chi-square test was applied to evaluate suture stages by sex and age groups., Results: Stage A was observed in only one 11-year-old girl. Stage B was present at all ages but was more prevalent in those less than 13 years of age. Stage C was the most prevalent in all evaluated ages. Stages D and E showed low prevalence rates. There were higher prevalences of the early stages of maturation in boys., Conclusions: The results of this study, which showed dominant prevalence of stage C, suggest that conventional, nonsurgical rapid maxillary expansion performed in patients over 15 years old is justified by a satisfactory prognosis when assessment of the sutural status indicates stage C., (Copyright © 2017 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Buccolingual inclinations of the maxillary permanent molars and intermolar widths increase with growth for Class I subjects. Changes for untreated Class II subjects have not yet been assessed. The aim of this study was to test the hypothesis that changes in palatal inclination of the maxillary molars and intermolar width throughout growth vary between Class I and Class II molar occlusions., Methods: Patients were selected from the Forsyth/Moorrees Twin Study. Dental models taken for 6 consecutive years of 55 untreated subjects (28 with Angle Class I and 27 with Angle Class II occlusion) were scanned. The images were superimposed on the palatal rugae, and the angle between a reference plane and the buccolingual inclination plane was used to calculate the buccolingual molar inclination at each time point. The distance between lingual groove points was used to calculate the intermolar width., Results: All molars showed increasing palatal inclinations over the 6 years. The change for each time interval was statistically significant. Class I subjects demonstrated significantly greater palatal inclination at each time point. The molar inclination changed by means of 4.99° for Class I subjects and 6.25° for Class II subjects. Intermolar width increased continuously (P <0.001) and was significantly greater (P <0.05) for Class I patients., Conclusions: These results suggest that palatal inclination of the maxillary permanent first molars occurs continuously between ages 9 and 14 years, with Class II subjects showing greater changes. The intermolar width increases steadily during this time, with Class II subjects having a narrower intermolar width and less change over time., (Copyright © 2017 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Head and neck skeletal anomalies or normal variants might predict the occurrence of palatally displaced impacted maxillary canines. Despite their clinical importance, studies in this regard are rare, especially when it comes to vertebral anomalies., Methods: This case-control study was performed on cephalographs of 35 orthodontic patients (11 male, 24 female) with palatally displaced canines (PDC) and 75 patients without them (29 male, 46 female). PDC were diagnosed on panoramic and lateral cephalographs and from clinical reports. The occurrence and severity of sella turcica bridge and the atlas ponticulus posticus, and deficiency of the posterior atlas arch were evaluated twice on lateral cephalographs. The associations between the occurrence and level of these skeletal anomalies and variations of PDC occurrence as well as additional correlations were assessed using multivariable and bivariate statistics (α = 0.05; β ≤0.2)., Results: The patients' mean age was 18.4 ± 1.9 years. In the control and patient groups, 23 (30.7%) and 21 subjects (60%) had sella turcica bridging, respectively (chi-square, P = 0.003). Ponticulus posticus was observed in 14 (18.7%) controls and 15 (42.9%) patients (chi-square, P = 0.007). Posterior atlas arch deficiency was observed in 4 (5.3%) controls and 5 (14.3%) patients (chi-square, P = 0.111). The presence of ponticulus posticus and sella turcica bridging might be associated with increased odds of PDC occurrence for about odds ratios of 3.1 and 3.5 times, respectively (binary logistic regression)., Conclusions: PDC is positively associated with the occurrence and severity of sella turcica bridging and ponticulus posticus. The association between PDC and posterior atlas arch deficiency was inconclusive., (Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Ciliopathies are pleiotropic human diseases resulting from defects of the primary cilium, and these patients often have cleft lip and palate. IFT88 is required for the assembly and function of the primary cilia, which mediate the activity of key developmental signaling pathways. Through whole exome sequencing of a family of three affected siblings with isolated cleft lip and palate, we discovered that they share a novel missense mutation in IFT88 (c.915G > C, p.E305D), suggesting this gene should be considered a candidate for isolated orofacial clefting. In order to evaluate the function of IFT88 in regulating craniofacial development, we generated Wnt1-Cre;Ift88fl/fl mice to eliminate Ift88 specifically in cranial neural crest (CNC) cells. Wnt1-Cre;Ift88fl/flpups died at birth due to severe craniofacial defects including bilateral cleft lip and palate and tongue agenesis, following the loss of the primary cilia in the CNC-derived palatal mesenchyme. Loss of Ift88 also resulted in a decrease in neural crest cell proliferation during early stages of palatogenesis as well as a downregulation of the Shh signaling pathway in the palatal mesenchyme. Importantly, Osr2KI-Cre;Ift88fl/flmice, in which Ift88 is lost specifically in the palatal mesenchyme, exhibit isolated cleft palate. Taken together, our results demonstrate that IFT88 has a highly conserved function within the primary cilia of the CNC-derived mesenchyme in the lip and palate region in mice and is a strong candidate as an orofacial clefting gene in humans., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Introduction: During adolescence, increasing interdigitation of the midpalatal suture increases resistance to rapid maxillary expansion (RME); this decreases its skeletal effect. In this study, we aimed at determining whether a novel measure of midpalatal suture maturity, the midpalatal suture density ratio, can be used as a valid predictor of the skeletal response to RME., Methods: The midpalatal suture density ratio, chronologic age, cervical vertebral maturation, and the stage of midpalatal suture maturation were assessed before treatment for 30 patients (ages, 12.9 ± 2.1 years) who underwent RME as part of comprehensive orthodontic treatment. Measurements on cone-beam computed tomography scans were used to determine the proportions of prescribed expansion achieved at the greater palatine foramina, the nasal cavity, and the infraorbital foramina., Results: There was a statistically significant negative correlation between the midpalatal suture density ratio and both the greater palatine foramina and the infraorbital foramina (r = -0.7877 and -0.3647, respectively; P <0.05). In contrast, chronologic age, cervical vertebral maturation, and stage of midpalatal suture maturation were not significantly correlated to any of the assessed measures of skeletal expansion (r range, -0.2209 to 0.0831; P >0.05)., Conclusions: The midpalatal suture density ratio has the potential to become a useful clinical predictor of the skeletal response to RME. Conversely, chronologic age, cervical vertebral maturation, and stage of midpalatal suture maturation cannot be considered useful parameters to predict the skeletal effects of RME., (Copyright © 2017 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Several mutations, located mainly in the MSX1 homeodomain, have been identified in non-syndromic tooth agenesis predominantly affecting premolars and third molars. We identified a novel frameshift mutation of the highly conserved C-terminal domain of MSX1, known as Msx homology domain 6 (MH6), in a Japanese family with non-syndromic tooth agenesis. To investigate the importance of MH6 in tooth development, Msx1 was targeted in mice with CRISPR/Cas system. Although heterozygous MH6 disruption did not alter craniofacial development, homozygous mice exhibited agenesis of lower incisors with or without cleft palate at E16.5. In addition, agenesis of the upper third molars and the lower second and third molars were observed in 4-week-old mutant mice. Although the upper second molars were present, they were abnormally small. These results suggest that the C-terminal domain of MSX1 is important for tooth and palate development, and demonstrate that that CRISPR/Cas system can be used as a tool to assess causality of human disorders in vivo and to study the importance of conserved domains in genes.

Retinoic acid (RA), the active derivative of vitamin A, is one of the major regulators of embryonic development, including the development of the epidermis, the limbs and the secondary palate. In the embryo, RA levels are tightly regulated by the activity of RA synthesizing and degrading enzymes. Aberrant RA levels due to genetic variations in RA metabolism pathways contribute to congenital malformations in these structures. In vitro and in vivo studies provide considerable evidence on the effects of RA and its possible role in the development of the epidermis, the limbs and the secondary palate. In conjunction with other regulatory factors, RA seems to stimulate the development of the epidermis by inducing proliferation and differentiation of ectodermal cells into epidermal cells. In the limbs, the exact timing of RA location and level is crucial to initiate limb bud formation and to allow chondrogenesis and subsequent osteogenesis. In the secondary palate, the correct RA concentration is a key factor for mesenchymal cell proliferation during palatal shelf outgrowth, elevation and adhesion, and finally to allow bone formation in the hard palate. These findings are highly relevant to understanding the mechanism of RA signalling in development and in the aetiology of specific congenital diseases., (Copyright © 2016 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.)

Abnormal regulation of Sonic hedgehog (Shh) signaling has been described in a variety of human cancers and developmental anomalies, which highlights the essential role of this signaling molecule in cell cycle regulation and embryonic development. Gas1 and Boc are membrane co-receptors for Shh, which demonstrate overlapping domains of expression in the early face. This study aims to investigate potential interactions between these co-receptors during formation of the secondary palate. Mice with targeted mutation in Gas1 and Boc were used to generate Gas1; Boc compound mutants. The expression of key Hedgehog signaling family members was examined in detail during palatogenesis via radioactive in situ hybridization. Morphometric analysis involved computational quantification of BrdU-labeling and cell packing; whilst TUNEL staining was used to assay cell death. Ablation of Boc in a Gas1 mutant background leads to reduced Shh activity in the palatal shelves and an increase in the penetrance and severity of cleft palate, associated with failed elevation, increased proliferation and reduced cell death. Our findings suggest a dual requirement for Boc and Gas1 during early development of the palate, mediating cell cycle regulation during growth and subsequent fusion of the palatal shelves.

Background: A new method of early neonatal cheiloplasty has recently been employed on patients having complete unilateral cleft lip and palate (cUCLP). We aimed to investigate (1) their detailed palatal morphology before surgery and growth during the 10 months after neonatal cheiloplasty, (2) the growth of eight dimensions of the maxilla in these patients, (3) the development of these dimensions compared with published data on noncleft controls and on cUCLP patients operated using later operation protocol (LOP; 6 months of age)., Methods: Sixty-six virtual dental models of 33 longitudinally evaluated cUCLP patients were analysed using metric analysis, a dense correspondence model, and multivariate statistics. We compared the palatal surfaces before neonatal cheiloplasty (mean age, 4 days) and before palatoplasty (mean age, 10 months)., Results: The palatal form variability of 10-month-old children was considerably reduced during the observed period thanks to their undisturbed growth, that is, the palate underwent the same growth changes following neonatal cheiloplasty. A detailed colour-coded map identified the most marked growth at the anterior and posterior ends of both segments. The maxilla of cUCLP patients after neonatal cheiloplasty had a growth tendency similar to noncleft controls (unlike LOP)., Conclusions: Both methodological approaches showed that early neonatal cheiloplasty in cUCLP patients did not prevent forward growth of the upper jaw segments and did not reduce either the length or width of the maxilla during the first 10 months of life., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Acquisition of cis-regulatory elements is a major driving force of evolution, and there are several examples of developmental enhancers derived from transposable elements (TEs). However, it remains unclear whether one enhancer element could have been produced via cooperation among multiple, yet distinct, TEs during evolution. Here we show that an evolutionarily conserved genomic region named AS3&#95;9 comprises three TEs (AmnSINE1, X6b&#95;DNA and MER117), inserted side-by-side, and functions as a distal enhancer for wnt5a expression during morphogenesis of the mammalian secondary palate. Functional analysis of each TE revealed step-by-step retroposition/transposition and co-option together with acquisition of a binding site for Msx1 for its full enhancer function during mammalian evolution. The present study provides a new perspective suggesting that a huge variety of TEs, in combination, could have accelerated the diversity of cis-regulatory elements involved in morphological evolution., Competing Interests: The authors have declared that no competing interests exist.

Background: There is a lack of evidence on the relationship between prematurity and palatal abnormalities. The aim of this study was to evaluate the incidence of palatal groove, the associated factors and the persistence time in preterm infants from birth to 24 months of age., Methods: The children's data, medical history and eating habits were collected using a questionnaire answered by the legal guardian and updated every dental visit. Natal and neonatal data were obtained from the medical records. During the orofacial examination, the presence or absence of a palatal groove was observed. In order to evaluate for associations between independent variables and the palatal groove, descriptive analyses and bivariate analyses were conducted using the Mann-Whitney, Pearson's chi-squared and Fisher's exact tests, when appropriate. The Poisson regression analysis was used to determine risk and protective factors for the occurrence of palatal groove. The significance level was 0.05. For the persistence of palatal groove, a survival analysis was used (Kaplan Meier method)., Results: Seventy-four preterm infants were monitored. Palatal groove occurred in n = 19 (25.7 %) and persisted for an average time of 12 months. Bivariate analysis showed a significantly higher occurrence of palatal groove in girls (68.4 % vs 40 % with non-occurrence of palatal groove) as well as in infants that stayed longer in the neonatal intensive care unit (NICU) (median 37 days vs 20 days), that did not have exclusive breastfeeding (94.7 % vs 69.1 %), were intubated (median five days vs one day) or used an orogastric tube (median 33 days vs 15 days). The quantitative data for 'NICU', 'intubation' and 'orogastric tube' were correlated and estimated as risk factors for palatal groove formation in the unadjusted Poisson regression analysis., Conclusions: Palatal groove occur transiently in approximately one quarter of preterm infants, especially in infants that stay longer in the NICU, are intubated or use an orogastric tube.

Objective: To measure the maxillary dentoskeletal and soft tissue changes of severely obstructed mouth breathing (MB) young children who had their mode of breathing normalized after adenotonsillectomy (T&A), in comparison with a matched group of severely obstructed untreated MB children (CG)., Methods: Seventy patients who had an Ear, Nose, and Throat examination (ENT), including flexible nasal endoscopy, to confirm the severe obstruction of the upper airways and the indication of T&A composed the sample. Cephalograms and dental casts were available from the patient's orthodontic records. Treatment group (TG) and CG included 35 children each. Groups were matched by gender (24 males and 11 females in each group), age (TG, 6.7 ± 1.8 years; CG, 6.9 ± 2.3 years), tooth development (TG, 13 primary dentition, 22 mixed dentition; CG, 14 primary dentition, 21 mixed dentition), and skeletal maturation status. Records were taken at baseline (T0) and 1-year after T&A (T1) for TG; while CG records were taken with a 1-year interval. Dentoskeletal measurements were performed in the lateral cephalograms, and dental casts were used to assess the palatal volume and occlusal changes., Results: TG showed a significant increase (503.3 mm(3), P < 0.001) in the palatal volume (10% of change), while CG palatal volume was stable. No dimensional occlusal changes were detected between T0 and T1 in both groups. Significant downward (point A, 2.1 mm; ANS, 2.1 mm) and forward displacements (point A, 0.7 mm; ANS, 1 mm) of the anterior region of the maxilla were observed in the TG, but CG presented only significant downward displacement (point A, 1.8 mm; ANS, 1.4 mm). The maxillary posterior region (PNS, PTM, and Molar) displaced downward in both groups (P < 0.05), however no sagittal change was found. The palatal plane inclination was stable in both groups., Conclusions: TG presented significant increase in the palatal volume and in the forward displacement of the maxilla. No other significant maxillary dentoskeletal changes were found., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Members of the Sox gene family play critical roles in many biological processes including organogenesis. We carried out comparative in situ hybridisation analysis of seventeen Sox genes (Sox1-14, 17, 18 and 21) during murine palatogenesis from initiation to fusion of the palatal shelves above the dorsal side of the tongue. At palatal shelf initiation (E12.5), the localized expression of six Sox genes (Sox2, 5, 6, 9, 12 and 13) was observed in the shelves, whereas Sox4 and Sox11 showed ubiquitious expression. During the down-growth of palatal shelves (E13.5), Sox4, Sox5, and Sox9 exhibited restricted expression to the interior side of the palatal shelves facing the tongue. Following elevation of the palatal shelves (E14.5), Sox2, Sox11 and Sox21 expression was present in the midline epithelial seam. We thus identify dynamic spatio-temporal expression of Sox gene family during the process of palatogenesis., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Acquisition of the lower jaw (mandible) was evolutionarily important for jawed vertebrates. In humans, syndromic craniofacial malformations often accompany jaw anomalies. The basic helix-loop-helix transcription factor Hand2, which is conserved among jawed vertebrates, is expressed in the neural crest in the mandibular process but not in the maxillary process of the first branchial arch. Here, we provide evidence that Hand2 is sufficient for upper jaw (maxilla)-to-mandible transformation by regulating the expression of homeobox transcription factors in mice. Altered Hand2 expression in the neural crest transformed the maxillae into mandibles with duplicated Meckel's cartilage, which resulted in an absence of the secondary palate. In Hand2-overexpressing mutants, non-Hox homeobox transcription factors were dysregulated. These results suggest that Hand2 regulates mandibular development through downstream genes of Hand2 and is therefore a major determinant of jaw identity. Hand2 may have influenced the evolutionary acquisition of the mandible and secondary palate.

Aim: To investigate the developmental process of palate morphology, including the alveolar ridge, in healthy infants for the predental period of 7 months from immediately after birth., Methods: The subjects were 32 healthy infants. Four or more dental casts were taken of each subject from immediately after birth until 7 months, for a total of 144 dental casts. Twelve characteristics were then measured in order to morphologically study the subjects' palate development. Principal component analysis (PCA) was performed to investigate morphological changes in the palatal vault., Results: The 12 characteristics were classified into either the alveolar ridge characteristics group, which determined the size of the alveolar ridge, or the palate characteristics group, which determined palate morphology, with each group showing different growth patterns. The characteristics of width and length increased with age in the alveolar ridge characteristics group; this correlation was maintained throughout the predental period. Meanwhile, in the palate characteristics group, the characteristics showed major developmental changes in the first 2 to 3 months after birth, but the changes were subsequently fewer from 3 to 7 months. The PCA of the palatal vault showed that the first principal component increased until 3 months but subsequently ceased to change., Conclusions: In predental infants, growth patterns of palate morphology differed according to their characteristics. There were major developmental changes in the palate during the first 3 months after birth. The study findings suggest that palate growth in the first half of the predental period may affect subsequent palate growth.

Introduction: Nonsyndromic cleft palate is a common birth defect (1:700) with a complex etiology involving both genetic and environmental risk factors. Nicotine, a major teratogen present in tobacco products, was shown to cause alterations and delays in the developing fetus., Methods: To demonstrate the postpartum effects of nicotine on palatal development, we delivered three different doses of nicotine (1.5, 3.0, and 4.5mg/kg/d) and sterile saline (control) into pregnant BALB/c mice throughout their entire pregnancy using subcutaneous micro-osmotic pump. Dams were allowed to deliver (~day 21 of pregnancy) and neonatal assessments (weight, length, nicotine levels) were conducted, and palatal tissues were harvested for morphological and molecular analyses, as well as transcriptional profiling using microarrays., Results: Consistent administration of nicotine caused developmental retardation, still birth, low birth weight, and significant palatal size and shape abnormality and persistent midline epithelial seam in the pups. Through microarray analysis, we detected that 6232 genes were up-regulated and 6310 genes were down-regulated in nicotine-treated groups compared to the control. Moreover, 46% of the cleft palate-causing genes were found to be affected by nicotine exposure. Alterations of a subset of differentially expressed genes were illustrated with hierarchal clustering and a series of formal pathway analyses were performed using the bioinformatics tools., Conclusions: We concluded that nicotine exposure during pregnancy interferes with normal growth and development of the fetus, as well results in persistent midline epithelial seam with type B and C patterns of palatal fusion., Implications: Although there are several studies analyzing the genetic and environmental causes of palatal deformities, this study primarily shows the morphological and large-scale genomic outcomes of gestational nicotine exposure in neonatal mice palate.The previous version was incorrect. New authors Ali Nawshad, Hasan Otu, Janki Sharma, and Elizabeth Sheldon have been included in this version; the funding and acknowledgement sections have been updated accordingly; the article title, some text, and one supplementary data file have been edited; and the corresponding author has been changed. The original corresponding author regrets these earlier errors., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Skulls are amongst the most informative documents of evolutionary history but a complex geometry, coupled with composite material properties and complicated biomechanics, have made it particularly challenging to identify mechanical principles guiding the skull's morphogenesis. Despite this challenge, multiple lines of evidence, for example the relationship between masticatory function and the evolution of jaw shape, nonetheless suggest that mechanobiology plays a major role in skull morphogenesis. To begin to tackle this persistent challenge, cellular, molecular and tissue-level analyses of the developing mouse palate were coupled with finite element modeling to demonstrate that patterns of strain created by mammalian-specific oral behaviors produce complementary patterns of chondrogenic gene expression in an initially homogeneous population of cranial neural crest cells. Neural crest cells change from an osteogenic to a chondrogenic fate, leading to the materialization of cartilaginous growth plate-like structures in the palatal midline. These growth plates contribute to lateral expansion of the head but are transient structures; when the strain patterns associated with suckling dissipate at weaning, the growth plates disappear and the palate ossifies. Thus, mechanical cues such as strain appear to co-regulate cell fate specification and ultimately, help drive large-scale morphogenetic changes in head shape.

Cleft palate is among the most common birth defects in humans. Previous studies have shown that Shh signaling plays critical roles in palate development and regulates expression of several members of the forkhead-box (Fox) family transcription factors, including Foxf1 and Foxf2, in the facial primordia. Although cleft palate has been reported in mice deficient in Foxf2, whether Foxf2 plays an intrinsic role in and how Foxf2 regulates palate development remain to be elucidated. Using Cre/loxP-mediated tissue-specific gene inactivation in mice, we show that Foxf2 is required in the neural crest-derived palatal mesenchyme for normal palatogenesis. We found that Foxf2 mutant embryos exhibit altered patterns of expression of Shh, Ptch1, and Shox2 in the developing palatal shelves. Through RNA-seq analysis, we identified over 150 genes whose expression was significantly up- or down-regulated in the palatal mesenchyme in Foxf2-/- mutant embryos in comparison with control littermates. Whole mount in situ hybridization analysis revealed that the Foxf2 mutant embryos exhibit strikingly corresponding patterns of ectopic Fgf18 expression in the palatal mesenchyme and concomitant loss of Shh expression in the palatal epithelium in specific subdomains of the palatal shelves that correlate with where Foxf2, but not Foxf1, is expressed during normal palatogenesis. Furthermore, tissue specific inactivation of both Foxf1 and Foxf2 in the early neural crest cells resulted in ectopic activation of Fgf18 expression throughout the palatal mesenchyme and dramatic loss of Shh expression throughout the palatal epithelium. Addition of exogenous Fgf18 protein to cultured palatal explants inhibited Shh expression in the palatal epithelium. Together, these data reveal a novel Shh-Foxf-Fgf18-Shh circuit in the palate development molecular network, in which Foxf1 and Foxf2 regulate palatal shelf growth downstream of Shh signaling, at least in part, by repressing Fgf18 expression in the palatal mesenchyme to ensure maintenance of Shh expression in the palatal epithelium.

We investigated the relationship between mouse taste bud development and innervation of the soft palate. We employed scanning electron microscopy and immunohistochemistry using antibodies against protein gene product 9.5 and peripherin to detect sensory nerves, and cytokeratin 8 and α-gustducin to stain palatal taste buds. At E14, nerve fibers were observed along the medial border of the palatal shelves that tracked toward the epithelium. At E15.5, primordial stages of taste buds in the basal lamina of the soft palate first appeared. At E16, the taste buds became large spherical masses of columnar cells scattered in the soft palate basal lamina. At E17, the morphology and also the location of taste buds changed. At E18-19, some taste buds acquired a more elongated shape with a short neck, extending a variable distance from the soft palate basal lamina toward the surface epithelium. At E18, mature taste buds with taste pores and perigemmal nerve fibers were observed on the surface epithelium of the soft palate. The expression of α-gustducin was demonstrated at postnatal day 1 and the number of pored taste buds increased with age and they became pear-shaped at 8 weeks. The percent of pored fungiform-like papillae at birth was 58.3% of the whole palate; this increased to 83.8% at postnatal day 8 and reached a maximum of 95.7% at 12 weeks. The innervation of the soft palate was classified into three types of plexuses in relation to taste buds: basal nerve plexus, intragemmal and perigemmal nerve fibers. This study reveals that the nerve fibers preceded the development of taste buds in the palate of mice, and therefore the nerve fibers have roles in the initial induction of taste buds in the soft palate., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Children with unoperated cleft palates have nearly normal growth of their faces whereas patients who have had early surgical repair often exhibit midfacial hypoplasia. Surgical repair is responsible for the underlying bone growth arrest but the mechanisms responsible for these surgical sequelae are poorly understood. We simulated the effect of cleft palate repair by raising a mucoperiosteal flap in the murine palate. Three-dimensional micro-CT reconstructions of the palate along with histomorphometric measurements, finite element (FE) modeling, immunohistochemical analyses, and quantitative RT-PCR were employed to follow the skeletal healing process. Inflammatory bone resorption was observed during the first few days after denudation, which destroyed the midpalatal suture complex. FE modeling was used to predict and map the distribution of strains and their associated stresses in the area of denudation and the magnitude and location of hydrostatic and distortional strains corresponded to sites of skeletal tissue destruction. Once re-epithelialization was complete and wound contracture subsided, the midpalatal suture complex reformed. Despite this, growth at the midpalatal suture was reduced, which led to palatal constriction and a narrowing of the dental arch. Thus the simple act of raising a flap, here mimicked by denuding the mucoperiosteum, was sufficient to cause significant destruction to the midpalatal suture complex. Although the bone and cartilage growth plates were re-established, mediolateral skeletal growth was nonetheless compromised and the injured palate never reached its full growth potential. These data strongly suggest that disruption of suture complexes, which have intrinsic growth potential, should be avoided during surgical correction of congenital anomalies., (Copyright © 2015. Published by Elsevier Inc.)

Introduction: The aim of this study was to analyze the diagnostic performance of the cervical vertebral maturation (CVM) method in estimating accurately the stages of maturation of the midpalatal suture., Methods: Cone-beam computed tomography (CBCT) images from 142 subjects (84 female, 58 male; mean age, 14.8 ± 9.7 years) were analyzed by 2 calibrated examiners to define, by visual analysis, the maturational stages of the cervical vertebrae and the midpalatal suture. These CBCT images were required by orthodontists and surgeons for diagnosis and treatment purposes. Positive likelihood ratios (LHRs) were calculated to evaluate the diagnostic performance of the CVM stages in identifying the maturational stages of the midpalatal suture., Results: Positive LHRs greater than 10 were found for several cervical vertebral stages (CSs), including CS1 and CS2 for the identification of midpalatal suture stages A and B, CS3 for the diagnosis of midpalatal suture stage C, and CS5 for the assessment of midpalatal suture stages D and E. These positive LHRs indicated large and often conclusive increases in the likelihood that the CVM stages were associated with specific stages of midpalatal suture maturation. At CS4, there were a moderate positive LHR for stage C and low positive LHRs for stages D and E., Conclusions: Most CVM stages can be used for the diagnosis of the stages of maturation of the midpalatal suture, so that CBCT imaging may not be necessary in these patients. In the postpubertal period, however, an assessment of the midpalatal suture maturation using CBCT images may be indicated in deciding between conventional rapid maxillary expansion and surgically assisted rapid maxillary expansion. On the other hand, if the CVM stage cannot be assessed, chronologic age may be a viable alternative to predict some midpalatal suture stages (particularly the early stages)., (Copyright © 2015 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The aim of this investigation was to study the relationship between third molar agenesis-including the number of ageneses-and craniofacial structure growth., Methods: We reviewed 305 clinical histories of patients treated at the Orthodontics Unit of the Faculty of Medicine and Dentistry at the University of Valencia in Spain. This included radiographic records of optimal quality. Of these, 40 patients who had agenesis of at least 1 third molar were included in the study group. A control group was formed with another 40 patients with all 4 third molars present. For both groups, a further criterion for inclusion was cone-beam computed tomography records. The cephalometric analysis was performed with NemoCeph 3D software (version 11.3.1.38; Nemotec, Madrid, Spain)., Results: The only significant differences between the 2 groups were in the total gonial angle and the upper gonial angle (P ≤0.05), both of which were smaller in the study group., Conclusions: Third molar agenesis is associated with a reduction in Jarabak's gonial angle and upper gonial angle, characteristic of patients with a more horizontal or brachyfacial skeletal pattern. No significant differences were found in other measurements., (Copyright © 2015 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Development of the human skull and face is a highly orchestrated and complex three-dimensional morphogenetic process, involving hundreds of genes controlling the coordinated patterning, proliferation and differentiation of tissues having multiple embryological origins. Craniofacial malformations that occur because of abnormal development (including cleft lip and/or palate, craniosynostosis and facial dysostoses), comprise over one-third of all congenital birth defects. High-throughput sequencing has recently led to the identification of many new causative disease genes and functional studies have clarified their mechanisms of action. We present recent findings in craniofacial genetics and discuss how this information together with developmental studies in animal models is helping to increase understanding of normal craniofacial development., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)