This body of work is concerned with the genetics of craniofacial morphology and specifically with that of the cranial sutures which form fibrous articulations between the calvarial bones. The premature fusion of these sutures, known as craniosynostosis, is a common developmental abnormality and has been extensively utilised here as a tool through which to study the genetics of suture morphogenesis and craniofacial diversity. Investigations began with a search for polymorphisms associated with normal variation in human craniofacial characteristics. Denaturing High-Performance Liquid chromatography was used to identify polymorphisms in two genes causative for craniosynostosis by analysing DNA from a large cohort of individuals from four ethnogeographic populations. A single nucleotide polymorphism in fibroblast growth factor receptor 1 was identified as being associated with variation in the cephalic index, a common measure of cranial shape. To further, and specifically, investigate the molecular processes of suture morphogenesis gene expression was compared between unfused and prematurely fusing/fused suture tissues isolated from patients with craniosynostosis. Two approaches, both utilising Affymetrix gene expression microarrays, were used to identify genes differentially expressed during premature suture fusion. The first was a novel method which utilised the observation that explant cells from both fused and unfused suture tissue, cultured in minimal medium, produce a gene expression profile characteristic of minimally differentiated osteoblastic cells. Consequently, gene expression was compared between prematurely fused suture tissues and their corresponding in vitro de-differentiated cells. In addition to those genes known to be involved in suture morphogenesis, a large number of novel genes were identified which were up-regulated in the differentiated in vivo state and are thus implicated in premature suture fusion and in vivo osteoblast differentiation. The second microarray study involved an extensive analysis of 16 suture tissues and compared gene expression between unfused (n=9) and fusing/fused sutures (n=7). Again, both known genes and a substantially large number of novel genes were identified as being differentially expressed. Some of these novel genes included retinol binding protein 4 (RBP4), glypican 3 (GPC3), C1q tumour necrosis factor 3 (C1QTNF3), and WNT inhibitory factor 1 (WIF1). The known functions of these genes are suggestive of potential roles in suture morphogenesis. Realtime quantitative RT PCR (QRT-PCR) was used to verify the differential expression patterns observed for 11 genes and Western blot analysis and confocal microscopy was used to investigate the protein expression for 3 genes of interest. RBP4 was found to be localised on the ectocranial surface of unfused sutures and in cells lining the osteogenic fronts while GPC3 was localised to suture mesenchyme of unfused sutures. A comparison between each unfused suture (coronal, sagittal, metopic, and lambdoid) demonstrated that gene expression profiles are suture-specific which, based on the identification of differentially expressed genes, suggests possible molecular bases for the differential timing of normal fusion and the response of each suture to different craniosynostosis mutations. One observation of particular interest was the presence of cartilage in unfused lambdoid sutures, suggesting a role for chondrogenesis in posterior skull sutures which have generally been thought to develop by intramembranous ossification without a cartilage precursor. Finally, the effects of common media supplements used in in vitro experiments to stimulate differentiation of calvarial suture-derived cells were investigated with respect to their ability to induce in vivo-like gene expression. The response to standard differentiation medium (ascorbic acid + β-glycerophosphate) with and without dexamethasone was measured by both mineralisation and matrix formation assays and QRT-PCR of genes identified in the above described microarray studies. Both media induced collagen matrix and bone nodule formation indicative of differentiating osteoblasts. However, the genes expression profiles induced by both media differed and neither recapitulated the levels and profiles of gene expression observed in vivo for cells isolated from both fused and unfused suture tissues. This study has implications for translating results from in vitro work to the in vivo situation. Significantly, the dedifferentiation microarray study identified differentially expressed genes whose products may be considered candidates as more appropriate osteogenic supplements that may be used during in vitro experiments to better induce in vivo-like osteoblast differentiation. This study has made a substantial contribution to the identification of novel genes and pathways involved in controlling human suture morphogenesis and craniofacial diversity. The results from this research will stimulate new areas of inquiry which will one day aid in the development of better diagnostics and therapeutics for craniosynostosis, and other craniofacial and more general skeletal abnormalities.

Esta tese é composta por três capítulos. O primeiro teve o objetivo de avaliar a acurácia e a semelhança da reconstrução facial forense computadorizada (RFFC) tridimensional realizada com softwares livres. As RFFC foram realizadas no programa Blender® a partir de tomografias do crânio, utilizando templates do MakeHuman®. A avaliação da acurácia foi feita no CloudCompare® que comparou a RFFC com a pele na tomografia, enquanto a avaliação da semelhança foi realizada no Picasa® utilizando fotografias dos participantes. Os resultados mostraram que do total de pontos que formam cada reconstrução, 63.20% a 73.67% apresentaram uma distância de -2,5 <= x <= 2,5mm entre a RFFC e a superfície da pele, enquanto a distância média variou entre -1,66 a 0,33mm. Duas das quatro reconstruções foram reconhecidas objetivamente pelo Picasa®. As RFFC realizadas utilizando esses softwares apresentam plausíveis níveis de acurácia e semelhança, portanto indicam valor para uso no campo forense. Os outros dois capítulos tiveram como objetivo avaliar a estabilidade métrica facial do mesmo indivíduo por meio da análise de fotografias tomadas em um intervalo de tempo de cinco anos. Trata-se de um estudo longitudinal realizado com fotografias frontais padronizadas de 666 indivíduos adultos divididos por faixa etária e sexo. Com o programa SAFF 2D foram marcados 32 pontos, cujas coordenadas foram utilizadas para calcular 40 medidas, sendo 20 horizontais e 20 verticais. Cada uma dessas medidas foi dividida pelo diâmetro da íris e assim foram obtidas razões iridianas. Os resultados mostraram que a maioria das razões não sofreu variação estatisticamente significante. As razões que tiveram maior variação foram aquelas da região do nariz e da boca. Quando se compara as faixas etárias entre si observa-se que a grande maioria das razões é diferente, mostrando a influência da idade nas dimensões faciais. Quando se compara a estabilidade dentro mesmo sexo observa-se que houve razões que diminuíram e outras aumentaram tanto no sexo feminino quanto no sexo masculino, enquanto outras variaram apenas em um dos sexos. Quando se compara a variação entre os sexos observa-se que a maioria das razões é diferente, mostrando o dimorfismo sexual das medidas faciais. A face passa por alterações métricas ao longo da vida em todas as faixas etárias, principalmente na região do nariz e boca, com maiores diferenças após os 60 anos. Além disso, algumas medidas faciais são mais influenciadas pelo sexo do que outras. Entretanto, a maioria das medidas levantadas se mantem relativamente estáveis dentro de um período de 5 anos tanto em relação ao sexo quanto a idade. / This thesis contains three chapters. The aim of the first chapter was to evaluate the accuracy and recognition level of three-dimensional (3D) computerized forensic craniofacial reconstruction (CCFR) performed in a blind test on open-source software using computed tomography data from live subjects. The CCFRs were completed using Blender® with 3D models obtained from the computed tomography data and templates from the MakeHuman® program. The evaluation of accuracy was carried out in CloudCompare®, by geometric comparison of the CCFR to the subject 3D face model (obtained from the CT data). A recognition level was performed using the Picasa® with a frontal standardized photography. The results were presented from all the points that form the CCFR model, with an average for each comparison between 63.20% and 73.67% with a distance -2.5 <= x <= 2.5 mm from the skin surface and the average distances were 1.66 to 0.33 mm. Two of the four CCFRs were correctly matched by the Picasa® tool. Free software programs are capable of producing 3D CCFRs with plausible levels of accuracy and recognition and therefore indicate their value for use in forensic applications. The other two chapters study the facial comparison and aimed to evaluate the facial metrical stability of an individual through photographs taken in a time interval of five years. It is a longitudinal study composed of standard frontal photographs of 666 adults divided by sex and age groups. By using the SAFF 2D® software, 32 landmarks were positioned, whose coordinates were used to calculate 40 measurements, 20 horizontal and 20 vertical. Each of these measurements was divided by iris diameter and thus iridian ratios were obtained. The results showed that most of the ratios did not suffer statistically significant variations. The ratios that had the greatest variation in the different age groups were those of the nose and mouth regions. When comparing the age groups with each other it is observed that the great majority of the reasons are different, showing the influence of age on the facial dimensions. When comparing stability with respect to sex, it was observed that there were ratios that decreased and others that increased in both sexes, while other ratios varied only in females or in males. When the sexes were compared, it was observed that the majority of the ratios were different, showing sexual dimorphism of the facial measures. The face undergoes metrical alterations throughout the life, mainly in the region of the nose and mouth, with the greatest differences seen in those who are aged 60 years and older. In addition, some facial measures are more influenced by sex than others. However, most of the measures raised have remained relatively stable within a period of five years in both sex and age groups.

- 1 - Abstract The intent of this thesis is to evaluate the differences in facial morphology of children between 3 and 15 years of age based on 3D facial models and cross-sectional data. Due to improper use of cross-sectional data for studying growth, the next part of the thesis is focused on the comparison of cross-sectional and longitudinal approaches in research. The longitudinal observation of facial developmental changes can be considered as actual growth. The cross-sectional database contains 839 3D facial models (397 boys, 442 girls). Three previously published longitudinal databases were used for comparison. Their age intervals were as follows: 3 to 6 years (12 boys, 14 girls), 6 to 12 years (15 boys, 18 girls), 12 to 15 years (23 boys, 22 girls). Geometric morphometric methods were used to analyse facial models (Coherent Point Drift - Dense Correspondance Analysis, Per Vertex T-Test and Principal Component Analysis). The results were visualized using superimposition colour maps, shell distance significance maps and their interlacing. When annual consecutive age intervals were used for cross-sectional data, we could not observe the fluency of differences in facial morphology between age categories, which we can observe during actual growth. When wider age intervals were used for cross-sectional...

Made available in DSpace on 2016-12-23T13:54:31Z (GMT). No. of bitstreams: 1 Nathalia Silveira Finck.pdf: 1726183 bytes, checksum: 40947138e78192ac535b3646b56a67ab (MD5) Previous issue date: 2013-08-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo dessa dissertação foi investigar as alterações temporomandibulares, craniofaciais e posturais associadas à respiração bucal, além de avaliar a relação entre sinais clínicos e sintomas de distúrbios respiratórios do sono (DRS) em escolares respiradores bucais, na faixa etária de 7 a 14 anos. Trata-se de um estudo caso-controle, com amostra de 147 escolares, sendo 73 com respiração bucal (RB) e 74 com respiração nasal (RN), avaliados através de anamnese, exame clínico e testes respiratórios. Os escolares diagnosticados como respiradores bucais responderam a um questionário sobre a autopercepção de sintomas de DRS na infância, com foco em problemas do sistema mastigatório, nasais e do sono. A presença de respiração bucal foi estatisticamente significativa para as seguintes alterações: ausência de selamento labial (OR= 29.70); desvio durante abertura da boca (OR= 24.63); palato atrésico (OR= 5.07); assimetria facial (OR= 5.06); índice de Mallampati III e IV (OR= 2.85); má oclusão Classe II (OR=2.67); hipertrofia de conchas nasais (OR= 2.19). Não houve diferença significativa entre os grupos para as alterações posturais. Nos escolares RB, problemas na ATM e acordar com dor de cabeça foram associados à má oclusão Classe II e à falta de selamento labial. Dor na nuca e torcicolo foram associados à sobremordida alterada e à anteriorização da cabeça. Problemas com o sono, sonolência diurna, acordar à noite, roncar e dormir de boca aberta foram associados à hipertrofia das tonsilas palatinas e ao índice de Mallampati obstrutivo. A chance de alterações temporomandibulares e craniofaciais ocorrerem em escolares com padrão de respiração bucal foi elevada. O aparecimento de sintomas de DRS na infância parece estar associado à persistência da respiração bucal e suas consequentes alterações craniofaciais, oclusais, posturais e nas vias aéreas superiores / The aim of this study was to investigate the temporomandibular, craniofacial and postural changes associated with mouth breathing and also evaluate the relationship between clinical signs and SDB symptoms in children 7 to 14 years of age. A case-control study with a sample of 147 children, 73 mouth breathers (MB) and 74 nasal breathers (NB), were evaluated by anamnesis, clinical examination and respiratory tests. The schoolchildren diagnosed as MB answered a questionnaire on self-perceived symptoms of SDB, focusing on the masticatory system, nasal and sleep problems. The presence of mouth breathing was statistically significant with the following changes: a lack of lip seal (OR=29.70), a deviation during the opening of the mouth (OR=24.63), an atresic palate (OR=5.07), a facial asymmetry (OR=5.06), an obstructive Mallampati scores (OR=2.85), a Class II malocclusion (OR=2.67) and a turbinate hypertrophy (OR=2.19). No significant difference in postural pattern was found between groups. In the MB group, TMJ problems and wake up with headache were associated with a Class II malocclusion and a lack of lip seal. Stiff neck or neck pain were associated with an altered overbite and a forward head position. Sleep problems, daytime sleepiness, waking at night, snoring and sleeping with the mouth open were associated with a tonsillar hypertrophy and obstructive Mallampati scores. The chances of occurrence of temporomandibular and craniofacial changes were high in the MB schoolchildren. The emergence of the SDB symptoms in childhood appears to be associated with the persistency of the mouth breathing and their consequent craniofacial, occlusal, postural and upper airway s abnormalities

Os defeitos da calota craniana causados por traumas severos, neoplasias, cirurgias ou deformidades congênitas representam um grande desafio para os cirurgiões. O uso de enxertia óssea autóloga continua sendo o método de tratamento padrão ouro, embora apresente morbidade na área doadora e seja considerado insuficiente para reconstrução de grandes defeitos. Recentemente, com o advento da bioengenharia tecidual, novas expectativas surgiram na regeneração óssea. O objetivo deste estudo foi desenvolver um modelo experimental em ratos para o estudo de deformidades craniofaciais e verificar se as células-tronco humanas provenientes de dentes decíduos seriam capazes de regenerar defeitos críticos em calota craniana de ratos não imunossuprimidos. Foram realizados dois defeitos ósseos de espessura total com diâmetro de 5 x 8 mm na região biparietal. O lado esquerdo foi preenchido com membrana de colágeno, enquanto o lado direito com membrana de colágeno associada a células-tronco humanas provenientes de dentes decíduos. Essas células foram caracterizadas previamente in vitro como células mesenquimais. A eutanásia dos animais foi realizada no 7º, 21º, 30º e 60º dia de pós-operatório e amostras de tecido ósseo foram extraídas para realização da análise histológica. A análise da presença de células humanas no novo osso formado foi confirmada através do estudo molecular. A linhagem de células-tronco humanas provenientes de dentes decíduos foi positiva para células-tronco mesenquimais e sua diferenciação em tecido ósseo também foi evidenciada in vitro. Foi observada a formação óssea após 21 dias de cirurgia nos dois lados, sendo o lado direito um osso mais maduro. A reação da cadeia de polimerase para DNA humano foi amplificada apenas no lado direito demonstrando que existiam células humanas nesse novo osso formado. O uso de células-tronco de dentes decíduos humanas em ratos não imunossuprimidos não evidenciou rejeição durante o período estudado. Os achados sugerem que o modelo experimental descrito poderá ser utilizado para o estudo dos defeitos ósseos cranianos em cirurgia craniofacial e que o uso de células-tronco humanas provenientes de dentes decíduos associado à membrana de colágeno parece representar uma importante estratégia para a reconstrução de tecidos ósseos e seu uso pode ser considerado uma opção para o reparo de grandes defeitos ósseos cranianos. / Repair of bone defects caused by severe trauma, resection of tumors, and congenital deformity remains a big challenge to surgeons. As a gold standard for the treatment of bone defects in clinic, autologous bone grafts are usually limited by considerable donor site mobility and available supply of tissue that can be harvested. Recently, tissue engineering has become a promising approach for bone regeneration. The main aim of this study is to create an experimental surgical protocol and evaluate the capacity of human dental pulp stem cells isolated from deciduous teeth, to reconstruct critical size cranial bone defects in nonimmunosuppressed rats. Bilateral 5 x 8 mm cranial full-thickness defects of parietal bone were created. The left side was supplied with collagen membrane only and the right side with collagen membrane and human dental pulp stem cells. Cells were used after in vitro characterization as mesenchymal cells. Animals were euthanized at 7, 21, 30 and 60 days postoperatively and cranial tissue samples were taken from the defects for histologic analysis. Analysis of the presence of human cells in the new bone was confirmed by molecular analysis. The human dental pulp stem cells lineage was positive for the four mesenchymal cell markers tested and showed osteogenic in vitro differentiation. The bone formation was observed 21 days after surgery on both sides, but a more mature bone was present in the right side. Human DNA was polymerase chain reaction-amplified only at the right side, indicating that this new bone had human cells. The use of human dental pulp stem cells in nonimmunosuppressed rats did not cause any graft rejection during this period. Our findings suggest that surgical protocol created may ultimately be used in experimental studies of cranial bone defects in craniofacial surgery and the use of human dental pulp stem cells together with collagen membrane seems to be a promising strategy for in vivo bone tissue reconstruction and their use might provide an option to repair human large cranial bone defects.

Craniofaciossinostose sindrômica (CS) é uma desordem congênita associada às mutações dos genes FGFR1, FGFR2 e FGFR3. Ocasiona uma fusão prematura das suturas craniofaciais, levando à deficiência no crescimento do crânio e terço médio da face. A severidade dos sintomas relacionados à hipertensão intracraniana, aparelho respiratório (síndrome da hipopneia-apneia do sono) e ocular (órbitas rasas, proptose ocular, exorbitismo e falta de proteção do globo ocular), são parâmetros para indicar o procedimento cirúrgico. A tomografia-3D é uma opção atual e pouco utilizada para avaliar os volumes orbitais e a magnitude dos avanços craniofaciais e seus vetores; esse método permite melhor definição das estruturas anatômicas sem distorção das imagens, promovendo uma avaliação adequada dos resultados. Os objetivos do presente estudo foram avaliar os volumes orbitais, os avanços craniofaciais e seus vetores nas osteotomias Le Fort III e Monobloco com uso de aparelhos distratores, a correlação entre os avanços craniofaciais e a variação dos volumes orbitais e determinar a eficácia dos avanços craniofaciais por meio da comparação com índices normais de volumes orbitais. No período de janeiro de 2001 a dezembro de 2012, foram selecionados 20 pacientes submetidos ao alongamento ósseo gradual do terço médio da face, divididos em dois grupos. Grupo LF (n = 9), submetidos à osteotomia Le Fort III e Grupo MB (n = 11), submetidos à osteotomia frontofacial monobloco. A avaliação constou da revisão de prontuários e exames tomográficos, em que foram mensurados o volume orbital pela segmentação de imagens no programa InVesalius e modelagem 3D no programa Magics, e a magnitude dos avanços craniofaciais, e seus vetores, foram medidos no programa Rhinoceros. Os resultados foram submetidos à análise estatística: teste t- Student e ANOVA. No Grupo MB, houve aumento de 8,94 mm3 e 9,84 mm3 para a variação do volume orbital, nas órbitas direita e esquerda, respectivamente; no Grupo LF: 5,70 mm3 e 5,77 mm3, respectivamente, para esses parâmetros. A resultante do avanço médio, para a órbita direita e esquerda no Grupo LF foi 11,36 ± 3,80 mm e 11,11 ± 3,45 mm, respectivamente; já no Grupo MB, foi 14,22 ± 4,12 mm e 14,48 ± 4,49 mm, respectivamente. A correlação entre a resultante do avanço e a variação dos volumes orbitais foi significante no Grupo LF e para a órbita esquerda no Grupo MB. Em conclusão, o procedimento cirúrgico foi eficaz, tanto para o Grupo LF como para o Grupo MB, pois, houve aumento estatisticamente significativo do volume orbital no período pós-operatório; não houve diferença estatisticamente significativa, na avaliação da simetria, entre os volumes orbitais pós-operatórios, direito e esquerdo. Ainda, não houve diferença estatisticamente significante entre os volumes orbitais pósoperatórios quando comparados aos índices de normalidade / Syndromic craniofacial synostosis (CS) is a congenital disorder, which is most often, an autosomal dominant mutation associated to FGFR1, FGFR2 and FGFR3 genes. It causes a premature fusion of craniofacial sutures, leading to deficiency in skull growth and middle-third of the face. The severity of the intracranial hypertension related, respiratory syndrome (hypopneaapnea) and eye (shallow orbits, ocular proptosis, exorbitism and lack of protection of the eyeball) symptoms, are parameters that indicate the surgical procedure. Distraction osteogenesis of the middle-third of the face is currently the chosen technique for the treatment of patients with Craniofacial Synostosis syndrome candidates for the advancement of the middle-third of the face. The 3D-tomography is a current option and seldom used for this purpose; this method allows better definition of anatomical structures without image distortion, providing a more accurate assessment of the results. The objectives of the present study, were to assess quantitatively the craniofacial advancement and its vectors through 3D tomographic images, the osteotomy Le Fort III and Monobloc with use of distractor devices; Also, to evaluate the correlation between the craniofacial advances and the variation of orbital volumes and determine the effectiveness of craniofacial advances through the comparison with normal levels of orbital volumes. From January 2001 to December 2012, 20 patients underwent distraction osteogenesis of the middle-third of the face, divided into two groups. LF group (n = 9) underwent a Le Fort III osteotomy and MB group (n = 11) underwent a monobloc frontalfacial osteotomy. The evaluation consisted of the reviewing of the records and tomographic exams; measured is the orbital volume by image segmentation on the InVesalius software and 3D modeling on Magics software and the magnitude of facial advances, and their vectors, were measured on Rhinoceros software. The results were subject to statistical analysis: t-Student test and ANOVA. In the MB group, there was an increase of 8.94 mm3 and 9.84 mm3 for the variation of the orbital volume, the right and left orbits, respectively; in Group LF: 5.70 mm3 and 5.77 mm3 for these parameters. The resulting of average advance, for right and left orbit in Group LF was of 11.36 ± 3.80 mm, 11.11 ± 3.45mm, respectively; as for group MB it was 14.22 ± 4.12 mm and 14.48 ± 4.49 mm, respectively. Correlation between the results from the progress and the variation of orbital volume was significant in Group LF and left orbit in MB Group. In conclusion, the surgical procedure was effective for both the LF group as for the MB Group because: there was a statistically significant increase in orbital volume on the postoperative period; there was no statistically significant difference in the evaluation of symmetry between the postoperative period left and right orbital volumes. Still, there was no statistically significant difference between the postoperative orbital volumes when compared to normal ranges

A expansão rápida da maxila assistida cirurgicamente (ERMAC) é o procedimento de eleição para o tratamento da deficiência transversa de maxila em pacientes esqueleticamente maduros. Controvérsias em relação ao procedimento ainda persistem principalmente relacionadas aos métodos de avaliação, técnica cirúrgica utilizada, morbidade, eficácia clínica e estabilidade de resultados. A utilização da tomografia computadorizada para a avaliação da eficiência da ERMAC é uma opção atual e ainda pouco divulgada. Consequentemente, os parâmetros anatômicos para a utilização deste método ainda não foram totalmente estabelecidos. Os objetivos do presente estudo foram avaliar a confiabilidade da metodologia proposta e as alterações esqueléticas maxilares decorrentes da ERMAC com a utilização do método de avaliação por tomografia computadorizada. No período de junho de 2004 a maio de 2005, 15 pacientes, sendo 10 do sexo feminino, foram submetidos a ERMAC. A técnica cirúrgica utilizada constou de osteotomia maxilar do tipo Le Fort I, sem osteotomia da sutura ptérigo-maxilar, associada a osteotomia sagital mediana do palato. O aparelho expansor utilizado foi do tipo Hyrax. Os pacientes foram submetidos a exames de tomografia computadorizada, em tomógrafos de múltiplos detectores, no pré-operatório e após 6 meses, utilizando-se metodologia idealizada para a avaliação específica deste procedimento cirúrgico. Para a determinação do padrão das alterações transversais e ântero-posteriores, medidas lineares e angulares foram realizadas diretamente na estação de trabalho dos aparelhos de tomografia. Através de imagens obtidas nas aquisições tomográficas axiais e em reconstruções coronais, as regiões anterior, média e posterior da maxila foram avaliadas separadamente. A área de secção transversa da maxila foi também calculada. Após a avaliação dos resultados observou-se que a confiabilidade do método foi estatisticamente comprovada. Um significativo aumento da área de secção transversa da maxila foi observado (p<0.05). Entretanto, o padrão de expansão transversa não se mostrou uniforme. O aumento das dimensões transversas nas regiões anterior e média foi estatisticamente comprovado. Não foi observada expansão transversa significativa na região posterior da maxila. Ao se avaliar a relação entre a abertura do parafuso expansor e a efetiva expansão esquelética da maxila, observou-se que esta foi menor que a abertura do parafuso, em termos absolutos. O aumento transverso relativo à expansão do parafuso foi estatisticamente maior nas regiões anterior e média da maxila do que em sua região posterior. Em conclusão, a avaliação por tomografia computadorizada para a análise das alterações esqueléticas da região maxilar, em pacientes submetidos à ERMAC é metodologia confiável e reprodutível. A expansão transversa da maxila decorrente do procedimento de ERMAC utilizado no presente estudo acarretou uma expansão não uniforme da maxila, com predomínio da expansão transversa nas regiões anterior e média. / Surgically assisted rapid palatal expansion is the procedure of choice for treating transverse maxillary deficiency in mature patients. Some controversies regarding surgically assisted rapid palatal expansion remain, mainly concerning technical aspects such as type and location of osteotomy sites, as well as surgical morbidity, clinical efficiency, and stability. The evaluation of transverse expansion is still a theme of discussion. On conventional anteroposterior radiographs, anatomical structures are superimposed, resulting in a high number of image artifacts, as well as hindering the tracing and evaluation. The use of computed tomography as a method of evaluating the efficiency of this procedure has not been widely reported. Consequently, few landmarks for use in evaluating maxillary expansion have been defined. The goals of the present study were to define parameters to assess skeletal changes after surgically assisted palatal expansion, to evaluate the reliability of the proposed method and to use computed tomography to assess those parameters. From June of 2004 to May of 2005, 15 patients underwent surgically assisted rapid palatal expansion (a modified Le Fort I maxillary osteotomy without pterygomaxillary separation, together with a sagital palatal osteotomy) according to a defined protocol, using a Hyrax appliance. To determine the pattern of transversal and anteroposterior expansion, linear and angular measurements were performed on multislice computed tomography, using computed software directly on the workstation. The anterior, intermediate and posterior portions of the maxilla were evaluated separately, using a specific method, in axial acquisition and coronal reconstructed views. The cross-sectional area of the maxilla was calculated to obtain general information about maxillary expansion. The reliability of the method was statistically confirmed. Significant maxillary overall expansion was observed. However, different patterns of expansion were seen in the three regions analyzed. In the anterior and intermediate portions of the maxilla, the increase in maxillary width was significantly greater than that observed in the posterior portion. The opening of the jackscrew was greater than skeletal expansion. Comparing jackscrew opening and transverse expansion, the same pattern of asymmetric expansion was verified. No change was observed in anteroposterior dimensions. The method of computed tomography evaluation is a useful tool for evaluation of surgically assisted rapid palatal expansion changes. The accurate evaluation of the postoperative changes was heavily dependent upon images acquired through computed tomography. An overall maxillary expansion was confirmed. However, transverse expansion of the maxilla achieved through surgically assisted rapid palatal expansion without pterygoid plate separation was less than uniform.

Introdução- Obstrução das vias aeríferas superiores associadas a mudanças nos padrões de sono, estão diretamente relacionados a problemas de crescimento e aprendizagem, o que interfere com a qualidade de vida das crianças com este quadro. Métodos de expansão maxilar já mostraram efeito favorável sobre a função respiratória. Aparelhos removíveis intra-orais têm sido usados no tratamento do ronco e apneia do sono, buscando reequilibrar a postura da mandíbula e da língua para melhorar a função respiratória. O objetivo deste trabalho é avaliar as dimensões da faringe, o crescimento craniofacial e os sintomas respiratórios obstrutivos em crianças com ronco e aumento das tonsilas faríngeas e palatinas em tratamento com um aparelho ortodôntico e ortopédico bucal. Métodos- Quarenta crianças de 6 a 9 anos de idade com tonsilas faríngeas e palatinas graus 3 e 4 e apresentando maxila atrésica e sobressaliência anterior foram divididos em dois grupos aleatórios: 24 pacientes tratados com o aparelho oral e 16 controles não tratados. As dimensões da faringe foram medidas por faringometria acústica. Cefalometria avaliou o crescimento facial, incluindo os valores relacionados com a apnéia do sono. Os pais preencheram questionários sobre os sintomas respiratórios da criança. Os pacientes foram reavaliados após 6 meses, em ambos os grupos. Resultados: A faringometria acústica confirmou um aumento volumétrico de 3,1 cm3 (d.p. ± 2,5) na faringe, no grupo de estudo e uma redução volumétrica de -1,2 cm3 (d.p. ± 1,3) no não tratado (p <0,001). A área mínima de colapsibilidade no grupo de estudo apresentou incremento de 1,1 cm2 (dp ± 0,2) para 1,3 cm2 (d.p. ± 0,2) e uma redução no grupo controle de 1,5 cm2 (dp ± 0,3) para 1,3 cm2 (d.p. ± 0,3) estatisticamente significante (p <0,001). A cefalometria comprovou crescimento craniofacial mais favorável no grupo de estudo, em comparação aos controles, incluindo os valores relacionados a apnéia do sono, como ANB, MMPA e H-ML (p <0,001) . O questionário de sintomas confirmou uma melhoria no padrão de respiração e sono no grupo tratado. Conclusão- As crianças que foram submetidos a esse protocolo de tratamento apresentaram aumento de dimensões da faringe, direção de crescimento mais favorável, e uma melhora na respiração e qualidade do sono / Introduction- Airway obstruction due to associated changes in sleep patterns are directly related to problems of growth and learning, which interfer with the quality of life for these children. Maxillary expansion methods have shown favorable effect on respiratory function. Intra-oral removable appliances have been used in the treatment of snoring and sleep apnea, seeking to rebalance the posture of the jaw and tongue to improve breathing function. The purpose of this thesis is evaluate the facial growth, pharyngeal dimensions and respiratory symptoms in snoring children with enlarged tonsils and adenoids under treatment with an orthodontic and orthopedic oral appliance. Methods- Forty snoring children ages 6 to 9 years old with tonsil and adenoid enlargement grades 3 and 4 and presenting constricted maxilla were divided into two randomized groups: 24 patients treated with the oral appliance and 16 untreated controls. Pharyngeal size was measured by acoustic pharyngometry. Cephalometry evaluated the facial growth including values related to sleep apnea. Pharyngeal size was measured by acoustic pharyngometry. Parents filled out questionnaires about their child\'s respiratory symptoms. Patients were re-evaluated after 6 months, in both groups Results- Acoustic pharyngometry confirmed a volumetric gain of 3.1 cm3 (s.d. ±2.5) in the pharynx at the study group and a volumetric reduction of -1.2 cm3 (s.d. ±1.3) at the untreated (p<0.001). The minimum area on collapsibility at the study group showed an increment from1.1 cm2 (s.d.±0.2) to 1.3 cm2 (s.d.±0.2) and a reduction on the control group from 1.5 cm2 (s.d.±0.3) to 1.3 cm2 (s.d.±0.3) statistically significant (p<0.001). Cephalometry showed a more favorable facial growth on the study group compared to controls, including values related to sleep apnea prediction such as ANB, MMPA and H-ML (p<0.001). The symptoms questionnaire confirmed an improvement on the breathing pattern at the group treated. Conclusions- Children who underwent this treatment protocol presented more favorable growth direction, enlargement of pharyngeal dimensions, and an improvement in breathing and sleep

Le syndrome de Goldenhar ou OAVS est une maladie du développement impliquant les deux premiers arcs branchiaux. Très hétérogène, elle est caractérisée par des anomalies des oreilles,des yeux et des vertèbres ainsi que par une microsomie hémifaciale. Des causes environnementales (exposition à l’Acide Rétinoïque (AR) durant la grossesse) et des causes génétiques (anomalies chromosomiques) ont été évoquées, mais aucun gène n’était directement associé à ce spectre. L’objectif de ce projet est donc d’identifier des gènes impliqués dans le spectre OAV. Des approches pangénomiques par séquençage nouvelle génération (exome,panels de gènes ciblés) ont été utilisées pour identifier des gènes candidats. L’identification de mutations dans MYT1 et l’inactivation transitoire de son l’homologue myt1a chez le poisson zèbre ont confirmé son rôle dans le développement cranio-facial et son implication dans l’OAVS. La validation fonctionnelle de ces mutations a été réalisée in vitro. Cible de la voie de l’Acide Rétinoïque (AR), MYT1 agit également comme répresseur des Récepteurs de l’AR permettant son rétrocontrôle négatif. Une approche toxicologique (traitements à l’AR de souris gestantes pendant une période clef du développement embryonnaire) a permis l’identification de protéines et de voies de signalisation dérégulées chez les embryons traités. L’étude de ces protéines modulées et notamment de celles déjà impliquées dans le développement cranio-facial tend à renforcer le lien entre AR et OAVS et offre des pistes intéressantes quant à l’identification de nouveaux gènes candidats pour ce syndrome, ces protéines pouvant être codées par des gènes potentiellement mutés chez des patients OAVS. / Goldenhar syndrome or Oculo-Auriculo-Vertebral Spectrum (OAVS) is a rare developmental disorder involving the first and the second pharyngeal arches. Extremely heterogeneous, it is characterized by hemifacial microsomia, asymmetric ears, ocular and vertebral abnormalities. Various etiologies have been suggested including environmental factors, especially embryonic Retinoic Acid (RA) exposure during pregnancy, and genetic causes (various chromosomal abnormalities). However, no gene had been formally implicated in this syndrome so far. The goal of this project is to identify genes involved in OAVS. Novel pangenomic approaches by Next generation Sequencing (Whole Exome Sequencing and Target genes Panel) were used to find new candidate genes. Identification of mutations in MYT1 and the transient knockdown experiments in zebrafish confirmed its implication in OAVS. Our in vitro studies provided functional characterization of these mutations and supported the link between MYT1 and RA signaling pathway. Thus, MYT1 is a target of RA but also acts as a repressor of RA Receptors and so, participates at the negative feedback. Toxicological approach was also performed by treatment of gestational mice by all-trans RA during a critical window of embryonic development. It led to a deregulation of proteins and to a modulation of cellular pathways in treated embryos. Studying the proteins whose expression is altered following the treatment, especially the proteins already involved in craniofacial development, could led to the identification of new candidate genes for OAVS and thus, may allow to better decipher the pathogenic mechanisms.

- 1 - Abstract The intent of this thesis is to evaluate the differences in facial morphology of children between 3 and 15 years of age based on 3D facial models and cross-sectional data. Due to improper use of cross-sectional data for studying growth, the next part of the thesis is focused on the comparison of cross-sectional and longitudinal approaches in research. The longitudinal observation of facial developmental changes can be considered as actual growth. The cross-sectional database contains 839 3D facial models (397 boys, 442 girls). Three previously published longitudinal databases were used for comparison. Their age intervals were as follows: 3 to 6 years (12 boys, 14 girls), 6 to 12 years (15 boys, 18 girls), 12 to 15 years (23 boys, 22 girls). Geometric morphometric methods were used to analyse facial models (Coherent Point Drift - Dense Correspondance Analysis, Per Vertex T-Test and Principal Component Analysis). The results were visualized using superimposition colour maps, shell distance significance maps and their interlacing. When annual consecutive age intervals were used for cross-sectional data, we could not observe the fluency of differences in facial morphology between age categories, which we can observe during actual growth. When wider age intervals were used for cross-sectional...