Identification of Novel Craniofacial Regulatory Domains Located far Upstream of SOX9 and Disrupted in Pierre Robin Sequence.

Mutations in the coding sequence of SOX9 cause campomelic dysplasia (CD), a disorder of skeletal development associated with 46,XY disorders of sex development (DSDs). Translocations, deletions, and duplications within a ∼2 Mb region upstream of SOX9 can recapitulate the CD-DSD phenotype fully or partially, suggesting the existence of an unusually large cis-regulatory control region. Pierre Robin sequence (PRS) is a craniofacial disorder that is frequently an endophenotype of CD and a locus for isolated PRS at ∼1.2-1.5 Mb upstream of SOX9 has been previously reported. The craniofacial regulatory potential within this locus, and within the greater genomic domain surrounding SOX9, remains poorly defined. We report two novel deletions upstream of SOX9 in families with PRS, allowing refinement of the regions harboring candidate craniofacial regulatory elements. In parallel, ChIP-Seq for p300 binding sites in mouse craniofacial tissue led to the identification of several novel craniofacial enhancers at the SOX9 locus, which were validated in transgenic reporter mice and zebrafish. Notably, some of the functionally validated elements fall within the PRS deletions. These studies suggest that multiple noncoding elements contribute to the craniofacial regulation of SOX9 expression, and that their disruption results in PRS.

Novel FGF8 mutations associated with recessive holoprosencephaly, craniofacial defects, and hypothalamo-pituitary dysfunction.

Context: Fibroblast growth factor (FGF) 8 is important for GnRH neuronal development with human mutations resulting in Kallmann syndrome. Murine data suggest a role for Fgf8 in hypothalamo-pituitary development; however, its role in the etiology of wider hypothalamo-pituitary dysfunction in humans is unknown.Objective: The objective of this study was to screen for FGF8 mutations in patients with septo-optic dysplasia (n = 374) or holoprosencephaly (HPE)/midline clefts (n = 47).Methods: FGF8 was analyzed by PCR and direct sequencing. Ethnically matched controls were then screened for mutated alleles (n = 480-686). Localization of Fgf8/FGF8 expression was analyzed by in situ hybridization in developing murine and human embryos. Finally, Fgf8 hypomorphic mice (Fgf8(loxPNeo/-)) were analyzed for the presence of forebrain and hypothalamo-pituitary defects.Results: A homozygous p.R189H mutation was identified in a female patient of consanguineous parentage with semilobar HPE, diabetes insipidus, and TSH and ACTH insufficiency. Second, a heterozygous p.Q216E mutation was identified in a female patient with an absent corpus callosum, hypoplastic optic nerves, and Moebius syndrome. FGF8 was expressed in the ventral diencephalon and anterior commissural plate but not in Rathke's pouch, strongly suggesting early onset hypothalamic and corpus callosal defects in these patients. This was consolidated by significantly reduced vasopressin and oxytocin staining neurons in the hypothalamus of Fgf8 hypomorphic mice compared with controls along with variable hypothalamo-pituitary defects and HPE.Conclusion: We implicate FGF8 in the etiology of recessive HPE and potentially septo-optic dysplasia/Moebius syndrome for the first time to our knowledge. Furthermore, FGF8 is important for the development of the ventral diencephalon, hypothalamus, and pituitary. (J Clin Endocrinol Metab 96: E1709-E1718, 2011)

Development of Craniofacial Bone Defect Reconstruction Implant Based on Fibre-Reinforced Composite with Photopolymerisable Resin Systems. Experimental studies in vitro and in vivo.

Reconstruction of defects in the craniomaxillofacial (CMF) area has mainly been based on bone grafts or metallic fixing plates and screws. Particularly in the case of large calvarial and/or craniofacial defects caused by trauma, tumours or congenital malformations, there is a need for reliable reconstruction biomaterials, because bone grafts or metallic fixing systems do not completely fulfill the criteria for the best possible reconstruction methods in these complicated cases. In this series of studies, the usability of fibre-reinforced composite (FRC) was studied as a biostable, nonmetallic alternative material for reconstructing artificially created bone defects in frontal and calvarial areas of rabbits. The experimental part of this work describes the different stages of the product development process from the first in vitro tests with resin-impregnated fibrereinforced composites to the in vivo animal studies, in which this FRC was tested as an implant material for reconstructing different size bone defects in rabbit frontal and calvarial areas. In the first in vitro study, the FRC was polymerised in contact with bone or blood in the laboratory. The polymerised FRC samples were then incubated in water, which was analysed for residual monomer content by using high performance liquid chromatography (HPLC). It was found that this in vitro polymerisation in contact with bone and blood did not markedly increase the residual monomer leaching from the FRC. In the second in vitro study, different adhesive systems were tested in fixing the implant to bone surface. This was done to find an alternative implant fixing system to screws and pins. On the basis of this study, it was found that the surface of the calvarial bone needed both mechanical and chemical treatments before the resinimpregnated FRC could be properly fixed onto it. In three animal studies performed with rabbit frontal bone defects and critical size calvarial bone defect models, biological responses to the FRC implants were evaluated. On the basis of theseevaluations, it can be concluded that the FRC, based on E-glass (electrical glass) fibres forming a porous fibre veil enables the ingrowth of connective tissues to the inner structures of the material, as well as the bone formation and mineralization inside the fibre veil. Bone formation could be enhanced by using bioactive glass granules fixed to the FRC implants. FRC-implanted bone defects healed partly; no total healing of defects was achieved. Biological responses during the follow-up time, at a maximum of 12 weeks, to resin-impregnated composite implant seemed to depend on the polymerization time of the resin matrix of the FRC. Both of the studied resin systems used in the FRC were photopolymerised and the heat-induced postpolymerisation was used additionally.

The role of fibroblast growth factor (FGF) and type beta transforming growth factot beta (TGF beta1, beta2 and beta3) during rat craniofacial development

Growth factors seem to be part of a complex cellular signalling language, in which individual growth factors are the equivalents of the letters that compose words. According to this analogy, informational content lies, not in an individual growth factor, but in the entire set of growth factors and others signals to which a cell is exposed. The ways in which growth factors exert their combinatorial effects are becoming clearer as the molecular mechanisms of growth factors actions are being investigated. A number of related extracellular signalling molecules that play widespread roles in regulating development in both invertebrates and vertebrates constitute the Fibroblast Growth Factor (FGF) and type beta Transforming Growth Factor ((TGF beta). The latest research literature about the role and fate of these Growth factors and their influence in the craniofacial bone growth ad development is reviewed

The role of fibroblast growth factor (FGF) and type beta transforming growth factot beta (TGF beta1, beta2 and beta3) during rat craniofacial development

Growth factors seem to be part of a complex cellular signalling language, in which individual growth factors are the equivalents of the letters that compose words. According to this analogy, informational content lies, not in an individual growth factor, but in the entire set of growth factors and others signals to which a cell is exposed. The ways in which growth factors exert their combinatorial effects are becoming clearer as the molecular mechanisms of growth factors actions are being investigated. A number of related extracellular signalling molecules that play widespread roles in regulating development in both invertebrates and vertebrates constitute the Fibroblast Growth Factor (FGF) and type beta Transforming Growth Factor ((TGF beta). The latest research literature about the role and fate of these Growth factors and their influence in the craniofacial bone growth ad development is reviewed

Características tomográficas da displasia fibrosa craniofacial: estudo retrospectivo de 14 casos

OBJETIVO: Apresentar as várias formas de displasia fibrosa craniofacial em um estudo revisional com 14 exames de tomografia computadorizada. MATERIAIS E MÉTODOS: Foram selecionados 14 casos de displasia fibrosa craniofacial atendidos no Serviço de Cirurgia de Cabeça e Pescoço e Otorrinolaringologia e no Serviço de Diagnóstico por Imagem do Hospital Heliópolis, São Paulo, SP, entre 1991 e 2002. Foram avaliados, pelos autores, três dos achados tomográficos mais relevantes: o número de ossos acometidos, a apresentação radiológica predominante e a uni/bilateralidade da doença no crânio. RESULTADOS: Observou-se predomínio do padrão misto em relação à forma de apresentação da doença. Foram vistos, ainda, acometimento contíguo de dois ou mais ossos e equivalência entre casos de acometimento craniofacial unilateral e bilateral, possivelmente por conta dos pacientes serem, em sua maioria, portadores de displasia fibrosa do tipo monostótica. CONCLUSÃO: O conhecimento das características tomográficas da displasia fibrosa craniofacial é de grande importância para que seja feito planejamento cirúrgico adequado e seguimento pós-tratamento.

Cephalometric analysis of craniofacial growth of a cohort of cleft lip and palate patients

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Adult craniofacial stem cells: sources and relation to the neural crest

During the process of development, neural crest cells migrate out from their niche between the newly formed ectoderm and the neural tube. Thereafter, they give rise not only to ectodermal cell types, but also to mesodermal cell types. Cell types with neural crest ancestry consequently comprise a number of specialized varieties, such as ectodermal neurons, melanocytes and Schwann cells, as well as mesodermal osteoblasts, adipocytes and smooth muscle cells. Numerous recent studies suggest that stem cells with a neural crest origin persist into adulthood, especially within the mammalian craniofacial compartment. This review discusses the sources of adult neural crest-derived stem cells (NCSCs) derived from the cranium, as well as their differentiation potential and expression of key stem cell markers. Furthermore, the expression of marker genes associated with embryonic stem cells and the issue of multi- versus pluripotency of adult NCSCs is reviewed. Stringent tests are proposed, which, if performed, are anticipated to clarify the issue of adult NCSC potency. Finally, current pre-clinical and clinical data are discussed in light of the clinical impact of adult NCSCs.

Holoprosencephaly and Holoprosencephaly-Like Phenotypes: Review of Facial and Molecular Findings in Patients From a Craniofacial Hospital in Brazil

Here we report on the clinical and genetic data for a large sample of Brazilian patients studied at the Hospital de Reabilitacao de Anomalas Craniofaciais-Universidade de Sao Paulo (HRAC-USP) who presented with either the classic holoprosencephaly or the holoprosencephaly-like (HPE-L) phenotype. The sample included patients without detected mutations in some HPE determinant genes such as SHH, GLI2, SIX3, TGIF, and PTCH, as well as the photographic documentation of the previously reported patients in our Center. The HPE-L phenotype has been also called of HPE ``minor forms"" or ""microforms,"" The variable phenotype, the challenge of genetic counseling, and the similarities to patients with isolated cleft lip/palate are discussed. (c) 2010 Wiley-Liss, Inc.

Craniometaphyseal Dysplasia With Severe Craniofacial Involvement Shows Homozygosity at 6q21-22.1 Locus

Craniotubular dysplasias (CTD) are a heterogeneous group of genetic disorders of skeletal development, whose clinical and etiological classification is still much debated. One of the most common form is the autosomal dominant craniometaphyseal dysplasia (CMD) which is associated with mutation in the ANKH gene. In the literature a few families are reported with CMD phenotype that suggest an autosomal recessive (AR) pattern of inheritance. A candidate locus at 6q21-22 has been mapped in a large inbred Brazilian family, but the gene of the recessive form is still unknown. Our data on a female patient with CMD phenotype, born from healthy first degree cousins and displaying homozygosity for polymorphic markers at the 6q21-22 locus, further support the existence of an AR CMD, expanding its clinical spectrum to a more severe phenotype. (C) 2011 Wiley-Liss, Inc.

Rotavirus gastroenteritis in a children's hospital specialized in craniofacial malformations

This study aimed to verify the relationship between acute diarrhea provoked by rotavirus and different indicators of craniofacial malformations. In the Hospital for Rehabilitation of Craniofacial Anomalies, 8,724 children with cleft lip and cleft palate were divided into the following groups: acute diarrhea and infection due to rotavirus (C1, n = 62), acute diarrhea (C2, n = 153) and without acute diarrhea (C3, n = 8,509). In C1, 29.03% of the cases consisted of hospital infections associated with the hospitalization period while 38.71% of the patients were aged less than six months. The percentage of children not having breastfed was significantly higher in acute diarrhea groups. Additionally, there was a seasonal prevalence of rotavirus infection between May and October. Finally, the present findings indicate that rotavirus is a predominant etiological agent for gastroenteritis in children with craniofacial malformations. Moreover, among infants younger than six months of age, type of craniofacial malformation, breastfeeding difficulty, socioeconomic level and longer hospitalization period appear to contribute to higher infection morbidity.

O significado do trabalho em equipe na reabilitação de pessoas com malformação craniofacial congênita

Este estudo objetivou compreender o significado do trabalho em equipe dos profissionais de reabilitação em anomalias craniofaciais. Realizou-se análise fenomenológica por contemplar a compreensão e interpretação do sentido considerando o sujeito. Entrevistou-se 12 profissionais de diferentes áreas, norteadas pela questão: O que significa para você trabalhar em equipe na reabilitação de anomalias craniofaciais? Resgatando-se os temas: Capacitação para o trabalho, Dificuldade para trabalho em equipe, Relação com paciente e família, Condições de trabalho e A inserção do profissional na equipe. A análise visou refletir o fenômeno engendrando convergências e divergências destacando a explicitação das diferenças e aprendizado contínuo.

Rapid Prototyping applied to surgical planning for correcting craniofacial malformations in wild animals. A case study of a brazilian fox

Veterinary surgery for treatment of wild animals is becoming an increasingly demanding task because it involves animals of different anatomy, many of them are already stressed and treatment must be performed to the highest standard in the minimum period of time. Craniofacial alterations may occur for three main reasons: genetic, functional or a combination of both. It is possible to modify the functional cause using intraoral devices like inclined plane. The treatment planning can be made based on virtual 3D models and rapid prototyping. An approximately 9 months old, 3.7 kg male Brazilian fox (Lycalopex vetulus) was referred to the Veterinary Hospital. Physical examination showed malocclusion with a deviation of the mandible to the right side. The virtual 3D model of the head was generated based on CT image data. The 3D models and rapid prototyping opened up new possibilities for the surgical planning and treatment of wild animals.

Retention Systems to Implant-Supported Craniofacial Prostheses

Osseointegrated implants in craniofacial reconstructions improve prostheses retention and stability and comfort and safety for a patient. According to biomechanical principles, the treatment success regarding osseointegration maintenance depends on an adequate surgical technique associated to a retention system that provides favorable tension distribution to implants. Furthermore, patient expectation, esthetics, function, and anatomic limitations must be evaluated during treatment planning. Therefore, the aims of this study were to present available retention systems to implant-supported craniofacial prosthesis and to highlight the advantages, indications, and limitations. A literature review was conducted through a MEDLINE search. Sixteen articles and 2 textbooks met the inclusion criteria and were included in the review. It was concluded that the success of craniofacial rehabilitation with implants depends on an adequate surgical technique and an adequate selection of a retention system.

Craniofacial Implants Success in Facial Rehabilitation

The use of craniofacial implants is an effective treatment for patients with deformities, burns, and cancer sequelae. The sites with the most successful implants are the auricular, nasal, and orbital regions. Furthermore, other factors can affect the implant longevity such as irradiated area, surgical technique, bone quality and quantity, macrostructure and microstructure of the implant, maintenance, and systemic factors.