This thesis further delineates the molecular genetic basis of a relatively common craniofacial condition, coronal craniosynostosis. It used whole-exome sequencing to identify novel disease genes in patients with non-syndromic coronal synostosis and negative genetic testing. Initially, 2 patients were identified with damaging, frameshift mutations in a gene not previously linked with craniosynostosis – Transcription Factor 12 (TCF12). A further intronic mutation was identified in a third patient. This gene encodes a transcription factor that dimerises with TWIST1, mutations of which cause Saethre-Chotzen syndrome, also associated with coronal synostosis. Screening 344 undiagnosed patients identified 35 further mutations, all with coronal synostosis with 14 cases arising de novo. This work was published and testing for TCF12-related craniosynostosis was translated clinically. Significant non-penetrance (60%) was identified in mutation-positive relatives and the genetic background was investigated. Firstly, analysis of parental origins of de novo mutations identified 6 of paternal origin and helped refine haplotype assignment. Secondly, haplotype analysis of TCF12-mutation carriers revealed modest correlation with phenotypic status, but this was insufficient to be useful in clinical testing. Thirdly, TCF12 haplotypes were analysed for association with non-syndromic coronal synostosis, but no significant association was found. Further exome sequencing revealed a de novo frameshift mutation in Transcription Factor 20 (TCF20) in a patient with coronal synostosis and autism, although the mutation only correlated with the latter phenotype. Analysis of 5 trios revealed a novel variant in myosin heavy chain 4 (MYH4) in 1 family, although its role in suture development is uncertain. Reviewing pooled exome data from 19 mutation-negative patients revealed no further disease genes. In summary, this thesis describes novel gene discovery, defines a new clinical entity and investigates genetic background of penetrant and non-penetrant individuals. Further exome sequencing identified another disease gene, a de novo mutation and compiled lists of damaging variants to allow future work.

Im Berliner Zentrum für Craniofaciale Fehlbildungen in der Klinik für Mund-, Kiefer- und Gesichtschirurgie- Klinische Navigation und Robotik der Charité Campus Virchow-Klinikum in Berlin wird der mobile CT Philips Tomoscan M in die Implantation von extraoralen kraniofazialen Knochenankern zur Epithesenfixierung eingebunden. Es stellt sich dabei die Frage nach der Ortstreue und Dimensionstreue von knöchernen Strukturen und extraoralen kraniofazialen Implantaten im CT. Ein Knochenphantom dient der Vermessung von Lochpaaren verschiedener Abstände, einer Knochenschraube, eines Abutments und eines Magneten unter variierten Scanparametern durch Werkzeuge der Scannerkonsole. Die Ortstreue von Löchern und Implantaten liegt in >99% im Intervall ±0,2mm. Fenstereinstellungen haben einen geringen Einfluss auf Lochabstände. Lochdurchmesser zeigen unter Anhebung des Window Level (WL) Korrelation (r2=0,776) und eine lineare Größenzunahme bei maßstabgerechter Skalierung, variieren aber wenig unter Variation der Window Width (WW). Alle Implantate werden zu groß abgebildet, außer in extremen Fenstereinstellungen. Implantatdimensionen nehmen bei maßstabgerechter Skalierung mit höheren WL linear ab. Die Variabilität der Darstellung nimmt mit dichterem Implantatmaterial ab. Kontaktflächen von Implantaten mit Gewebe sind im CT nicht beurteilbar. / In the Berlin Centre for Craniofacial Malformations at the Clinic for Oromaxillofacial Surgery of the Charité, Campus Virchow- Clinic in Berlin, a mobile CT Scanner Philips Tomoscan M is integrated into the process of extraoral bone anchor implantation for prosthesis fixture. With implementation of the scanner, accuracy of localisation and dimensional accuracy of bony structures and implants have been questioned. A bone phantom is used for measuring distances between holes, hole diameters and dimensions of an implant fixture, an abutment and a magnet with measuring tools of the scanner under variation of scan parameters. Localisation accuracy of holes and implants is found in >99% within ±0.2mm. Window settings have little influence on hole distances. Diameters show linear growth at standardized scaling and correlation with window level augmentation (r2=.776). Little Variation of diameters is found with variation of window width. All Implants are magnified. Implant dimensions diminish with window level augmentation at standardized scaling. Variability of dimension decreases with more radio opaque implant material. Implant contact with tissues cannot be assessed in CT scans.

Orientador: Fausto Viterbo de Oliveira Neto / Resumo: As deformidades craniofaciais decorrentes de traumas, ressecções de tumores ou malformações congênitas são freqüentes na prática médica e o tratamento destas necessitam de cirurgia reparadora, com técnicas especializadas e profissionais qualificados para corrigir os defeitos e proporcionar melhor qualidade de vida, aprimorar a fala, respiração, mastigação e deglutição. Há diversas técnicas descritas para corrigir os defeitos ósseos, cada uma com vantagens e desvantages, escolhidas de acordo com o tipo de deformidade. Este estudo avaliou a formação óssea em um retalho tubular vascularizado, gáleo-periostal, enriquecido com o uso de pó de osso, plasma rico em plaquetas, células-tronco mesenquimais e ácido hialurônico, em coelhos, que tenha capacidade de substituir o enxerto ósseo nas reconstruções, principalmente nos defeitos faciais. No estudo, utilizou-se 98 coelhos divididos em doze grupos, submetidos à cirurgia para confecção do retalho em calota craniana. Foram realizados retalhos tubulares com o periósteo voltado para dentro e preenchidos com pó de osso, plasma rico em plaquetas (PRP), células-tronco mesenquimais (CTM) e ácido hialurônico. O Grupo 1 não foi manipulado. No Grupo 2 foi realizado o retalho tubular e mantido vazio. O Grupo 3 teve o retalho preenchido com pó de osso, no Grupo 4 o retalho foi mantido vazio. O Grupo 5 teve o retalho preenchido com PRP. No Grupo 6 o retalho foi preenchido com PRP e pó de osso. O Grupo 7 foi preenchido com CTM. O Grupo 8 teve o ... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

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No. of bitstreams: 1 brock_rs_dr_bot.pdf: 138052465 bytes, checksum: f5b517e489dbadd6cf6b83fbef881ecd (MD5) Previous issue date: 2017-01-31 / As deformidades craniofaciais decorrentes de traumas, ressecções de tumores ou malformações congênitas são freqüentes na prática médica e o tratamento destas necessitam de cirurgia reparadora, com técnicas especializadas e profissionais qualificados para corrigir os defeitos e proporcionar melhor qualidade de vida, aprimorar a fala, respiração, mastigação e deglutição. Há diversas técnicas descritas para corrigir os defeitos ósseos, cada uma com vantagens e desvantages, escolhidas de acordo com o tipo de deformidade. Este estudo avaliou a formação óssea em um retalho tubular vascularizado, gáleo-periostal, enriquecido com o uso de pó de osso, plasma rico em plaquetas, células-tronco mesenquimais e ácido hialurônico, em coelhos, que tenha capacidade de substituir o enxerto ósseo nas reconstruções, principalmente nos defeitos faciais. No estudo, utilizou-se 98 coelhos divididos em doze grupos, submetidos à cirurgia para confecção do retalho em calota craniana. Foram realizados retalhos tubulares com o periósteo voltado para dentro e preenchidos com pó de osso, plasma rico em plaquetas (PRP), células-tronco mesenquimais (CTM) e ácido hialurônico. O Grupo 1 não foi manipulado. No Grupo 2 foi realizado o retalho tubular e mantido vazio. O Grupo 3 teve o retalho preenchido com pó de osso, no Grupo 4 o retalho foi mantido vazio. O Grupo 5 teve o retalho preenchido com PRP. No Grupo 6 o retalho foi preenchido com PRP e pó de osso. O Grupo 7 foi preenchido com CTM. O Grupo 8 teve o retalho preenchido com CTM e pó de osso. O Grupo 9 teve o retalho preenchido com CTM e PRP. No Grupo 10, o retalho tubular foi preenchido com PRP, CTM e pó de osso. O Grupo 11 foi mantido vazio e o Grupo 12 foi preenchido com ácido hialurônico. Os resultados foram avaliados através de métodos de imagem e avaliação histológica. Os resultados demonstraram que, no modelo experimental utilizado, os grupos com apenas periósteo, isto é, retalho tubular vazio, apresentaram formação óssea pequena e irregular. No grupo com PRP também houve a formação óssea irregular e imatura. Quando o PRP foi associado ao pó de osso houve uma formação mais regular e organizada. O grupo com célula-tronco mesenquimal também apresentou formação óssea, com características teciduais organizadas, próprias do tecido ósseo maduro. Quando associada ao pó de osso e ao PRP, as características histológicas apresentaram-se com tecido organizado, regular, maduro com células bem formadas e organizadas. O uso de materiais com fatores de crescimento celular ósseo melhoram a qualidade e organização do tecido neoformado. Quanto maior o número de fatores de enriquecimento usados, melhores foram os resultados quanto a qualidade tecidual neoformada. / Craniofacial deformities caused by traumas, tumor ressections or congenital malformation are frequent in medical practice, and their treatment with reconstructive surgeries are common, especially in plastic surgery, which aim to provide the patients with better quality of life and functional improvement of speach, breathing, chewing and swallowing. Many different techniques are described to correct bone defects. They have advantages and disadvantages, chosen according to the type of deformity. This study evaluated a vascularized galeal and periosteum flap filled with bone fragments, platelet rich plasma, mesenchymal stem cells and hyaluronic acid, using rabbits, which could possibily substitute the bone graft in reconstructive surgery, especially for facial defects. It was an experimental study, with 98 rabbits divided into twelve groups, submitted to a surgical procedure to construct a calvaria flap. A tubular flap with the periosteum inside was constructed and filled with bone fragments, platelet rich plasma (PRP), mesenchymal stem cells (MSC) and hyaluronic acid. Group 1 was not manipulated. In Group 2, the tubular flap was maintained empty. Group 3 had the flap filled with bone fragments, in Grupo 4 the flap was maintained empty. Group 5 had the flap filled with PRP. In Group 6 the flap was filled with PRP and bone fragments. The Group 7 was filled with MSC. Group 8 had the flap filled with MSC and bone fragments. The Group 9 had the flap filled with MSC and PRP. In Group 10, the tubular flap was filled with PRP, MSC and bone fragments.The Group 11 was maintained empty and Group 12 was filled with hyaluronic acid. The results were evaluated using image methods and histological analysis. The results demonstrated that, in the experimental model used, the groups with only periosteum, this is the empty tubular flap, presented small and irregular bone formation. In the group with PRP, it also had irregular and imature bone formation. When the PRP was associated to bone fragments it had a more regular and organized formation. The group with mesenchymal stem cell also presented bone formation, with organized tissue characteristics, proper of mature osseous tissue. When associated to bone fragments and PRP, the histological characteristics presented organized, regular, mature tissue with organized and well formed cells. The use of materials with osseous cellular growth factors improves the quality and organization of the neoformed tissue. The more enrichment factor used, the better the neoformed tissue quality result was.