Made available in DSpace on 2014-07-29T15:07:34Z (GMT). No. of bitstreams: 1 Dissert-Danilo Ferreira Rodrigues.pdf: 452744 bytes, checksum: 0d66f9b822194b140597d873ca8c4a8c (MD5) Previous issue date: 2011-02-25 / Peritoneal adhesions are highly important in clinical and surgical practice because of the potential to cause sequelae such as chronic abdominal pain, infertility and bowel obstruction; thus, preventing the formation of this process becomes a challenge for surgeons. Due to biocompatibility, bioabsorption, healing and antimicrobial properties, chitosan is a polymer which has aroused interest of researchers studying the prevention of peritoneal adhesions. The objective of this research was to determine whether the intraperitoneal implantation of polypropylene mesh coated with a film of chitosan/polyethylene glycol is effective for the prevention of adhesions in rats with induced abdominal defects. We sought to further characterize the tissue reactions in acute and chronic phases of the inflammation process, and the tissue repair processes triggered by this biomaterial and the polypropylene mesh without coating. Defect of about 1cm was made in the abdominal wall, with reference to anatomical aponeurosis of the fascia of the rectus abdominis and obliquus externus abdominis muscles, followed by the implantation of polypropylene meshes coated with chitosan / polyethylene glycol (QP Group, n= 12) and uncoated polypropylene meshes (PP group, n= 12). Six animals from each group were euthanized at four and 45 days after implantation and the involvement of the mesh area by the adhesion, the type of adhesion, the inflammation process and tissue repair, and the quantification and characterization of collagen fibers were evaluated. The average of the meshes involved by adhesions in the QP and PP groups was 39.088% and 84.024%, respectively, and the structure involved in abdominal adhesions in the QP group usually consisted of the omentum around the implant, while in the PP group, adhesions were usually on the surface of the mesh. After four days of surgery, the amount of mononuclear cells was lower in QP, but the amount of polymorphonuclear cells, fibrin and collagen fibers was in greater quantity in this group; besides, the later variable showed better spatial organization. After 45 days of biomaterials implantation, there was a reduction in the amount of inflammatory cells, and spatial organization of collagen fibers in the QP group remained strong. It is concluded that the meshes coated with the film based on chitosan/polyethylene glycol did not prevent adhesion formation, but significantly minimized the seriousness of this process. In the acute phase of inflammation and tissue repair, the foreign body reaction was less intense in the group receiving the coated polypropylene mesh, but the tissue reactions caused by the studied biomaterials were similar to the chronic phase. The mesh coated with film-based chitosan/polyethylene glycol stimulated higher production and better orientation of collagen type I in the acute phase of inflammation and tissue repair, and better orientation of collagen fibers in the chronic phase of this process. / As aderências peritoniais possuem grande importância na prática clínico-cirúrgica pela possibilidade de causarem sequelas como dor abdominal crônica, infertilidade e obstrução intestinal, portanto a prevenção do desenvolvimento deste processo torna-se um desafio para os cirurgiões. Devido às propriedades de biocompatibilidade, bioabsorção, cicatrizante e antimicrobiana, a quitosana é um polímero que tem despertado interesse no estudo da prevenção de aderências peritoniais. O objetivo com esta pesquisa foi verificar se o implante intraperitonial de telas de polipropileno revestidas com o filme de quitosana/polietilenoglicol é eficiente na prevenção de aderências em ratos com defeitos abdominais induzidos, além de caracterizar as reações teciduais nas fases aguda e crônica do processo de inflamação e reparo tecidual desencadeadas por este biomaterial, e a tela de polipropileno sem revestimento. Foram realizados defeitos na parede abdominal de aproximadamente 1cm, tendo como referência anatômica a aponeurose das fáscias externas dos músculos reto abdominal e oblíquo abdominal externo, seguida da implantação de telas de polipropileno revestidas com quitosana/polietilenoglicol (Grupo QP, n=12) e telas de polipropileno sem revestimento (Grupo PP, n=12). Seis animais de cada grupo foram submetidos à eutanásia aos quatro e 45 dias após o implante, e foram avaliadas a área de envolvimento da tela pela aderência, o tipo de aderência, o processo de inflamação e reparo tecidual, além da quantificação e tipificação de fibras colágenas. A média das telas envolvidas pelas aderências no grupo QP foi de 39,088% e de 84,024% no grupo PP e a estrutura abdominal envolvida nas aderências no grupo QP geralmente consistiam pelo omento ao redor do implante, no grupo PP, as aderências geralmente estavam na superfície da tela. Após quatro dias do procedimento cirúrgico, a quantidade de células mononucleares foi menor no grupo QP, porém a quantidade de células polimorfonucleares, fibrina e fibras colágenas estavam em maior quantidade nesse grupo, além desta última variável apresentar melhor organização espacial. Após 45 dias da implantação dos biomateriais, houve a redução na quantidade de células inflamatórias, e a organização espacial das fibras colágenas no grupo QP foi permaneceu acentuada. Conclui-se que as telas revestidas com o filme à base de quitosana/polietilenoglicol não preveniram a formação de aderências, porém minimizaram significativamente a gravidade deste processo. Na fase aguda do processo de inflamação e reparo tecidual, a reação de corpo estranho foi menos intensa no grupo que recebeu a tela de polipropileno com revestimento, porém as reações teciduais provocadas pelos biomateriais analisados foram similares na fase crônica. A tela revestida com o filme a base de quitosana/polietilenoglicol estimulou a maior produção e melhor orientação de fibras colágenas tipo I na fase aguda do processo de inflamação e reparação tecidual, e melhor orientação de fibras colágenas na fase crônica deste processo.

[pt] A informação sobre o posicionamento de objetos estranhos no interior do corpo humano é essencial para a sua eficiente remoção cirúrgica. Entretanto, os métodos convencionalmente utilizados não fornecem informação suficiente sobre a localização do objeto metálico para garantia de sucesso cirúrgico. No presente trabalho foi desenvolvido um sistema automatizado para mapear a densidade de fluxo magnético estático produzido por corpos ferromagnéticos posicionados em variados graus de liberdade 3D, utilizando um sensor de baixo custo, baseado no fenômeno da magnetoimpedância gigante (GMI - Giant Magnetoimpedance), que detecta somente campos magnéticos variantes no tempo. Assim, as medições automatizadas foram realizadas com a amostra em movimento a uma velocidade constante. Por meio de modelagens computacionais do campo magnético gerado foi possível reproduzir o comportamento da densidade de fluxo magnético gerado por uma fonte de campo magnético como a agulha retilínea utilizada nas medições in vitro. O software considerou as características do sensor GMI utilizado e a condição de medição com a fonte magnética em movimento. Os resultados da simulação foram validados por meio de comparações com os resultados experimentais, possibilitando a solução do problema direto com a caracterização da configuração espacial da densidade de fluxo magnético para variados posicionamentos da fonte magnética em relação ao sensor magnético GMI. Com a validação dos resultados simulados, os mesmos podem ser empregados no desenvolvimento de procedimento para solução do problema inverso de imageamentos clínicos utilizando o sensor GMI de baixo custo, limitado a medições magnéticas variantes no tempo, realizados para detecção e posicionamento de corpos estranhos que geram campos magnéticos estáticos. / [en] Information about the positioning of foreign objects inside the human body is essential for its efficient surgical removal. However, the methods conventionally used do not provide sufficient information on the location of the metallic object to guarantee surgical success. In the present work, an automated system was developed to map the static magnetic flux density produced by ferromagnetic bodies positioned in varying degrees of 3D freedom, using a low-cost sensor based on the giant magnetoimpedance phenomenon (GMI - Giant Magnetoimpedance), which detects only time-varying magnetic fields. Thus, automated measurements were performed with the sample moving at a constant speed. Through computational modeling of the generated magnetic field, it was possible to reproduce the behavior of the magnetic flux density generated by a magnetic field source, such as the straight needle used in in vitro measurements. The software considered the GMI sensor s characteristics and the measurement condition with the magnetic source in motion. The simulation results were validated through comparisons with the experimental results, enabling the solution of the direct problem with the characterization of the spatial configuration of the magnetic flux density for various magnetic source positions in relation to the GMI magnetic sensor. With the validation of the simulated results, they can be used in the development of a procedure to solve the inverse problem of clinical imaging using the low-cost GMI sensor, limited to time-varying magnetic measurements, performed for the detection and positioning of foreign bodies that generate static magnetic fields.

[pt] Este trabalho apresenta a estratégia híbrida das abordagens Quality by Design e Failure Mode and Effects Analysis para aplicação no desenvolvimento de tecnologia biomédica. A qualidade e a gestão de riscos precisam evoluir com o avanço do desenvolvimento farmacêutico e tecnológico na saúde, permitindo um maior controle dos processos de fabricação de cada dispositivo biomédico. A Análise de Modo e Efeito de Falha (FMEA) tem sido amplamente utilizada na saúde para garantir a qualidade dos produtos fabricados. Ela se dedica a prevenir eventos ou incidentes adversos, identificando falhas para priorizar o risco e otimizando recursos, para uma melhor qualidade em dispositivos ou equipamentos médicos. A Qualidade por Projeto (QbD), por sua vez, tem sido aplicada na indústria farmacêutica para garantir a qualidade dos produtos desde a fase de desenvolvimento. Os resultados obtidos com a estratégia híbrida desenvolvida determinaram os atributos da qualidade e os parâmetros do processo mais críticos para aplicação no desenvolvimento de sistema de medição para localização de corpo estranho metálico em pacientes. A abordagem QbD-FMEA contribuiu para a determinação mais adequada e abrangente dos elementos críticos mais relevantes e associadas ações preventivas do que o emprego dessas sistemáticas isoladamente. / [en] This work presents the hybrid strategy of Quality by Design and Failure Mode and Effects Analysis approaches for application in biomedical technology development. Quality and risk management need to evolve with the advancement of pharmaceutical and technological development in healthcare, allowing greater control of the manufacturing processes of each biomedical device. Failure Mode and Effect Analysis (FMEA) has been widely used in healthcare to ensure the quality of manufactured products. It is dedicated to preventing adverse events or incidents, identifying failures to prioritize risk, and optimizing resources for better medical devices or equipment quality. In turn, Quality by Design (QbD) has been applied in the pharmaceutical industry to ensure product quality from the development phase. The results obtained with the developed hybrid strategy determined the most critical quality attributes and process parameters for application in developing a measurement system for metallic foreign body location in patients. The hybrid QbD-FMEA approach contributed to a more adequate and comprehensive determination of the most relevant critical elements, associating with preventive actions, than the isolated use of these approaches.

In der vorliegenden Arbeit wurden retrospektiv 66 Fälle stattgehabter Fremdkörperaspirationen aus den kinderchirurgischen Akten und Ambulanzakten der Jahre 1997 bis 2008 des Universitätsklinikums Leipzig rekonstruiert und im Hinblick auf die nachfolgend genannten Kriterien statistisch ausgewertet: Geschlecht und Alter der Kinder, Indikation zur Bronchoskopie, Einweisungsart, Dauer bis zur Diagnosestellung, endoskopische Sichtbarkeit des Fremdkörpers, Entfernungsschritte, erforderliche Nachbeatmung, Anhalt durch Röntgenbefund, auskultatorischer Anhalt, anamnestischer Anhalt, Eigen- oder Fremdanamnese, das Vorhandensein von Husten und die Dauer bis zur Entlassung aus dem Krankenhaus. Die Kriterien wurden standardisiert und mittels des Statistikprogramms SPSS 18 ausgewertet. Anhand der ausgewerteten Daten wurde ein Algorithmus zur innerklinischen Vorgehensweise bei Verdacht auf Fremdkörperaspiration erstellt, welcher eine interdisziplinäre Arbeitsrichtlinie (Standard Operation Procedure, SOP) für andere Kinderzentren liefern kann. Das geschärfte Bewusstsein für die Häufigkeit einer Fremdkörperaspiration vor allem bei Kindern und die damit verbundene problemorientierte Diagnostik und Therapie soll mit einer einschlägigen Arbeitsrichtlinie in Zukunft mehr Kindern als bisher zunutze gemacht werden.

Thanks to pioneering scientists and clinicians, prosthetic devices that are controlled by intra-cortical electrodes recording one's 'thoughts' are a reality today, and no longer merely in the realm of science fiction. However, widespread clinical use of implanted electrodes is hampered by a lack of reliability in chronic recordings, independent of the type of electrodes used. The dominant hypothesis has been that astroglial scar electrically impedes the electrodes. However, recent studies suggest that the impedance changes associated with the astroglial scar are not high enough to interfere significantly impair neural recordings. Furthermore, there is a time delay between when scar electrically stabilizes and when neural recordings fail (typically >1 month lag), suggesting that scar, per se, does not cause chronic recording unreliability. In this study, an alternative hypothesis was tested in a rat model, namely, that chronic inflammation surrounding microelectrodes causes a local neurodegenerative state. Chronic inflammation was varied in three ways: 1) stab wound control, 2) age-matched control, and 3) inter-shank spacing of a multishank electrode. The results of this study suggest that chronic inflammation, as indicated by activated microglia and reactive astrocytes, is correlated with local neurodegeneration, marked by neuron cell death and dendritic loss. Surprisingly, axonal pathology in the form of hyperphosphorylation of the protein Tau (the hallmark of many tauopathies, including Alzheimer's Disease) was also observed in the immediate vicinity of microelectrodes implanted for 16 weeks. Additionally, work is presented on a fast, non-invasive method to monitor the astrocytic response to intra-cortical electrodes using electrical impedance spectroscopy. This work provides a non-invasive monitoring tool for inflammation, albeit an indirect one, and fills a gap which has slowed the development of strategies to control the inflammatory tissue response surrounding microelectrodes and thereby improve the reliability of chronic neural recordings. The results of these experiments have significance for the field of neuroengineering, because a more accurate understanding of why recordings fail is integral to engineering reliable solutions for integrating brain tissue with microelectrode arrays.

Bei angeborenen Herzfehlern, die bei 1 bis 1,2 % aller Lebendgeburten auftreten und so-mit die häufigste behandlungsbedürftige Organfehlbildung darstellen, kommen regelhaft kardiovaskuläre Implantate im Rahmen der chirurgischen bzw. interventionellen Therapie zum Einsatz. Hierzu zählen u. a. Shunts, Patches und Okkluder, die aus verschiedenen Implantatmaterialien hergestellt werden. Das Ziel dieser Arbeit bestand darin – basierend auf histologischen Untersuchungen – Unterschiede bzw. Gemeinsamkeiten bezüglich der Biokompatibilität nicht-metallischer Implantatmaterialien zu prüfen, da eine bewusste Materialauswahl kardiovaskulärer Implantate zur Therapie angeborener Herzfehler zu besseren Langzeitergebnissen der Implantate beitragen kann. Untersucht wurden Implantate, die im Rahmen von Korrekturoperationen entnommen wurden und anschließend im Forschungslabor für Pädiatrische Kardiologie und Intensivmedizin der Universitätsmedizin Göttingen ausgewertet wurden: Shunts aus PTFE (n = 21, durchschnittliche Implantationszeit: 18 Monate), Patches aus PTFE (n = 13, durchschnittliche Implantationszeit: 247 Monate) und Polyester (n = 4, durchschnittli¬che Implantationszeit: 321 Monate) sowie Okkluder aus PTFE (n = 3, durchschnittliche Implantationszeit: 74 Monate), Polyester (n = 9, durchschnittliche Implantationszeit: 30 Monate) und PVA (n = 2, durchschnittliche Implantationszeit: 23 Monate). Zur Herstellung histologischer Präparate wurden metallhaltige Implantate (Okkluder) sowie solche mit bereits makroskopisch sichtbarer Verkalkung in Methylmethacrylat-Kunstharz eingebettet und anschließend gesägt und geschliffen, sodass sie lichtmikroskopisch ausgewertet werden konnten. Die anderen Implantate wurden in Paraffin eingebettet und geschnitten. Neben konventionellen Färbungen zur Übersicht und Darstellung von Verkalkungen wurden immunhistochemische Färbungen eingesetzt. Unabhängig vom Implantatmaterial konnte regelhaft eine endothelialisierte und neovaskularisierte Pseudointima, hauptsächlich am ehesten aus Myofibroblasten und Fibroblasten bestehend, dargestellt werden. Das im Implantatmaterial neu gebildete Gewebe bestand hauptsächlich aus Fibroblasten und war neovaskularisiert. Implantatassoziierte, chronische Entzündungsreaktionen – getragen durch Makrophagen und Lymphozyten – sowie Fremdkörperreaktionen – getragen durch FKR – waren bei den Polyester- und PVA-Implantaten stärker ausgeprägt als bei den PTFE-Implantaten. Verkalkungen in Pseudointima- und Implantatgewebe wurden bei den Polyester-Implantaten ab einer Implantationszeit von 3 Jahren und 4 Monaten, bei den PTFE-Implantaten ab einer Implantationszeit von 5 Jahren und 10 Monaten beobachtet. Die durch Polyester hervorgerufene, stärker ausgeprägte Entzündungsreaktion ist als Ursache der zu einem früheren Zeitpunkt einsetzenden Verkalkung von Polyester-Implantaten anzusehen. Während bei den Polyester-Implantaten häufig eher ungleichmäßig verteilte und unregelmäßig geformte, punktförmige Verkalkungen bis hin zu kleinen Kalkaggregaten in Pseudointima- und Implantatgewebe vorhanden waren, wiesen die PTFE-Implantate zumeist gleichmäßige, großflächig-konfluierende Verkalkungen auf. Es konnte gezeigt werden, dass bei Implantaten, die Polyester- oder PTFE-Anteile enthal¬ten, mittelfristig mit der Entwicklung von lokalen Verkalkungen zu rechnen ist, die im Langzeitverlauf zu Komplikationen führen können. Dies muss bei der Implantatauswahl beachtet werden. Möglicherweise kann in Zukunft durch die Entwicklung neuartiger Materialien eine Verminderung der Verkalkungstendenz, zum Beispiel durch Biodegra¬dierbarkeit des Implantatmaterials, erreicht werden.

L’immuno-isolation des îlots de Langerhans est proposée comme moyen d’effectuer des transplantations sans prise d’immunosuppresseurs par le patient. Cette immuno-isolation, par l’entremise d’une microcapsule composée d’alginate et de poly-L-lysine (microcapsule APA), protège le greffon d’une éventuelle attaque du système immunitaire du receveur grâce à sa membrane semi-perméable. Cette membrane empêche le système immunitaire du receveur de pénétrer la microcapsule tout en laissant diffuser librement les nutriments, le glucose et l’insuline. Avant l’application de cette technique chez l’humain, quelques défis doivent encore être relevés, dont la biocompatibilité de ce système. La biocompatibilité fait ici référence à la biocompatibilité du biomatériau utilisé pour la fabrication des microcapsules, l’alginate, mais aussi la biocompatibilité des microcapsules reliée à leur stabilité. En effet, il a été remarqué que, lors d’implantation in vivo de microcapsules fabriquées avec de l’alginate non purifiée, ceci induisait un phénomène nommé Réaction de l’Hôte contre la Microcapsule (RHM). De plus, il est connu que la stabilité des microcapsules APA peut influencer leur biocompatibilité puisqu’une microcapsule endommagée ou brisée pourrait laisser s’échapper les cellules du greffon chez le receveur. Nous croyons qu’une compréhension des processus d’initiation de la RHM en fonction de l’efficacité des procédés de purification d’alginate (et donc des quantités de contaminants présents dans l’alginate) ainsi que l’augmentation de la stabilité des microcapsules APA pourront améliorer la biocompatibilité de ce dispositif, ce que tente de démontrer les résultats présentés dans cette thèse. En effet, les résultats obtenus suggèrent que les protéines qui contaminent l’alginate jouent un rôle clé dans l’initiation de la RHM et qu’en diminuant ces quantités de protéines par l’amélioration des procédés de purification d’alginate, on améliore la biocompatibilité de l’alginate. Afin d’augmenter la stabilité des microcapsules APA, nous décrivons une nouvelle technique de fabrication des microcapsules qui implique la présence de liaisons covalentes. Ces nouvelles microcapsules APA réticulées sont très résistantes, n’affectent pas de façon négative la survie des cellules encapsulées et confinent les cellules du greffon à l’intérieur des microcapsules. Cette dernière caractéristique nous permet donc d’augmenter la biocompatibilité des microcapsules APA en protégeant le receveur contre les cellules du greffon. / Islet of Langerhans inmmunoisolation is proposed as a way to avoid the use of immunosuppressive drugs after transplantation. Microcapsules, the immuno-isolating device, are composed of alginate and poly-L-lysine and the protection of the graft is granted by a semi-permeable membrane. This membrane allows small molecules to freely diffuse within the microcapsule, such as nutrients, glucose and insulin while protecting the graft against the host immune system. Biocompatibility is one of the challenges that must be addressed before the successful clinical application of this device. Microcapsules biocompatibility is related, first, to the biocompatibility of alginate, the polymer used to made microcapsules and second, to the in vivo stability of these microcapsules. In facts, it is well know that the use of an unpurified alginate containing many foreign contaminants to make microcapsules induce the host reaction against microcapsule (HRM). Moreover, damaged or broken microcapsules can allow the dissemination of cells from the encapsulated graft, activating the host immune system. We believe that a better understanding of the initiation processes of the HRM in terms of alginate purification efficacy to remove contamination as well as an improve microcapsule stability will increase microcapsules biocompatibility. Results reported in this thesis suggest that foreign proteins found in alginate are playing a key role in the initiation of HRM and that the reduction of these foreign proteins, by the improvement of alginate purification processes, improves microcapsules biocompatibility. In order to increase microcapsules stability, we also described and characterized an innovative type of microcapsules which involve covalent bonds. These covalently cross-linked microcapsules were found to by highly resistant and stable. The novel fabrication process of these microcapsules was not harmful for the encapsulated cell survival and was also found to confine the graft inside the microcapsules. This characteristic enables us to increase microcapsules biocompatibility by the protection of the host from the encapsulated cells.

L’immuno-isolation des îlots de Langerhans est proposée comme moyen d’effectuer des transplantations sans prise d’immunosuppresseurs par le patient. Cette immuno-isolation, par l’entremise d’une microcapsule composée d’alginate et de poly-L-lysine (microcapsule APA), protège le greffon d’une éventuelle attaque du système immunitaire du receveur grâce à sa membrane semi-perméable. Cette membrane empêche le système immunitaire du receveur de pénétrer la microcapsule tout en laissant diffuser librement les nutriments, le glucose et l’insuline. Avant l’application de cette technique chez l’humain, quelques défis doivent encore être relevés, dont la biocompatibilité de ce système. La biocompatibilité fait ici référence à la biocompatibilité du biomatériau utilisé pour la fabrication des microcapsules, l’alginate, mais aussi la biocompatibilité des microcapsules reliée à leur stabilité. En effet, il a été remarqué que, lors d’implantation in vivo de microcapsules fabriquées avec de l’alginate non purifiée, ceci induisait un phénomène nommé Réaction de l’Hôte contre la Microcapsule (RHM). De plus, il est connu que la stabilité des microcapsules APA peut influencer leur biocompatibilité puisqu’une microcapsule endommagée ou brisée pourrait laisser s’échapper les cellules du greffon chez le receveur. Nous croyons qu’une compréhension des processus d’initiation de la RHM en fonction de l’efficacité des procédés de purification d’alginate (et donc des quantités de contaminants présents dans l’alginate) ainsi que l’augmentation de la stabilité des microcapsules APA pourront améliorer la biocompatibilité de ce dispositif, ce que tente de démontrer les résultats présentés dans cette thèse. En effet, les résultats obtenus suggèrent que les protéines qui contaminent l’alginate jouent un rôle clé dans l’initiation de la RHM et qu’en diminuant ces quantités de protéines par l’amélioration des procédés de purification d’alginate, on améliore la biocompatibilité de l’alginate. Afin d’augmenter la stabilité des microcapsules APA, nous décrivons une nouvelle technique de fabrication des microcapsules qui implique la présence de liaisons covalentes. Ces nouvelles microcapsules APA réticulées sont très résistantes, n’affectent pas de façon négative la survie des cellules encapsulées et confinent les cellules du greffon à l’intérieur des microcapsules. Cette dernière caractéristique nous permet donc d’augmenter la biocompatibilité des microcapsules APA en protégeant le receveur contre les cellules du greffon. / Islet of Langerhans inmmunoisolation is proposed as a way to avoid the use of immunosuppressive drugs after transplantation. Microcapsules, the immuno-isolating device, are composed of alginate and poly-L-lysine and the protection of the graft is granted by a semi-permeable membrane. This membrane allows small molecules to freely diffuse within the microcapsule, such as nutrients, glucose and insulin while protecting the graft against the host immune system. Biocompatibility is one of the challenges that must be addressed before the successful clinical application of this device. Microcapsules biocompatibility is related, first, to the biocompatibility of alginate, the polymer used to made microcapsules and second, to the in vivo stability of these microcapsules. In facts, it is well know that the use of an unpurified alginate containing many foreign contaminants to make microcapsules induce the host reaction against microcapsule (HRM). Moreover, damaged or broken microcapsules can allow the dissemination of cells from the encapsulated graft, activating the host immune system. We believe that a better understanding of the initiation processes of the HRM in terms of alginate purification efficacy to remove contamination as well as an improve microcapsule stability will increase microcapsules biocompatibility. Results reported in this thesis suggest that foreign proteins found in alginate are playing a key role in the initiation of HRM and that the reduction of these foreign proteins, by the improvement of alginate purification processes, improves microcapsules biocompatibility. In order to increase microcapsules stability, we also described and characterized an innovative type of microcapsules which involve covalent bonds. These covalently cross-linked microcapsules were found to by highly resistant and stable. The novel fabrication process of these microcapsules was not harmful for the encapsulated cell survival and was also found to confine the graft inside the microcapsules. This characteristic enables us to increase microcapsules biocompatibility by the protection of the host from the encapsulated cells.

In der vorliegenden Arbeit wurden retrospektiv 66 Fälle stattgehabter Fremdkörperaspirationen aus den kinderchirurgischen Akten und Ambulanzakten der Jahre 1997 bis 2008 des Universitätsklinikums Leipzig rekonstruiert und im Hinblick auf die nachfolgend genannten Kriterien statistisch ausgewertet: Geschlecht und Alter der Kinder, Indikation zur Bronchoskopie, Einweisungsart, Dauer bis zur Diagnosestellung, endoskopische Sichtbarkeit des Fremdkörpers, Entfernungsschritte, erforderliche Nachbeatmung, Anhalt durch Röntgenbefund, auskultatorischer Anhalt, anamnestischer Anhalt, Eigen- oder Fremdanamnese, das Vorhandensein von Husten und die Dauer bis zur Entlassung aus dem Krankenhaus. Die Kriterien wurden standardisiert und mittels des Statistikprogramms SPSS 18 ausgewertet. Anhand der ausgewerteten Daten wurde ein Algorithmus zur innerklinischen Vorgehensweise bei Verdacht auf Fremdkörperaspiration erstellt, welcher eine interdisziplinäre Arbeitsrichtlinie (Standard Operation Procedure, SOP) für andere Kinderzentren liefern kann. Das geschärfte Bewusstsein für die Häufigkeit einer Fremdkörperaspiration vor allem bei Kindern und die damit verbundene problemorientierte Diagnostik und Therapie soll mit einer einschlägigen Arbeitsrichtlinie in Zukunft mehr Kindern als bisher zunutze gemacht werden.:1. Einführung 1.1. Einleitung 1.2. Anatomische Grundlagen a) Allgemeiner Aufbau der Atemwege/luftleitenden Strukturen b) Physiologische Besonderheiten beim Kind c) Ätiologie des Aspirationsereignisses d) Ablauf/Störungen des Schluckaktes 2. Aufgabenstellung und Zielsetzung 3. Materialien und Methode 3.1. Materialien 3.2. Methode a) Statistische Analyse b) Literatur 4. Ergebnisse 5. Diskussion 6. Zusammenfassung 7. Literaturverzeichnis 8. Anlagen 8.1. Lebenslauf 8.2. Danksagung 8.3. Eigenständigkeitserklärung

Introduction: Les implants dans le système nerveux périphérique (SNP) peuvent potentiellement restaurer les capacités sensorielles et motrices chez les patients avec des amputations des membres supérieures. Cependant, la réaction à un corps étrangers affecte significativement la fonction à long-terme et la biocompatibilité de ces systèmes avec le temps. Le dendrimère (DND) et la Poly-D-Lysine (PDL) sont deux polymères synthétiques qui peuvent potentiellement améliorer la performance de ces implants. Pour cette étude, notre objectif est de déterminer si ces polymères peuvent promouvoir la formation d’éléments présynaptiques sur des surfaces synthétiques in vivo dans un modèle animal. Méthodes: Pour l’étude in vivo, nous avons utilisé un modèle d’écrasement du nerf sciatique chez le rat. Des billes enduites de DND et PDL et contrôle ont été injectées dans le nerf sciatique aux sites d’écrasement et 5 mm distaux au site d’écrasement. Après 4, 6 et 8 semaines, les nerfs ont été retirés et marqués avec des anticorps spécifiques au neurofilament et à la synaptophysine. Nous avons ensuite compté le nombre d’éléments présynaptiques retrouvant sur la surface de chaque bille pour toutes les conditions. Pour l’étude de l’électrode, deux électrodes ont été implantées dans le nerf sciatique du rat. Nous avons ensuite effectué des enregistrements nerveux à chaque semaine, et le potentiel d’action dans le nerf a été mesuré en variant uniquement la largeur de l’impulsion. Résultats: L’étude in vivo a démontré que les billes enduites de DND pouvaient promouvoir une accumulation significative de synaptophysine sur leurs surfaces comparé aux billes contrôles de 4 à 8 semaines. À 4 semaines, les billes dans la condition DND avaient également une accumulation de synaptophysine significativement supérieure à celles dans la condition PDL pour le site distal à l’écrasement. L’étude de l’électrode a démontré que les deux électrodes pouvaient stimuler et acquérir des signaux nerveux du nerf sciatique jusqu’à 1 et 2 semaines respectivement avant de ne plus fonctionner. Conclusion: Les résultats de notre étude suggèrent que DND possède une propriété à promouvoir la synaptogenèse qui est supérieure à PDL in vivo et que notre modèle d’électrode peut être utilisé pour évaluer la stabilité du signal des implants SNP. / Background: Implants in the peripheral nervous system (PNS) can potentially restore sensory feedback, improve motor control and alleviate phantom-limb pain in upper-limb amputees. However, nervous system implants have poor long-term function and biocompatibility when implanted into the body due to foreign body reaction. Dendrimer (DND) and Poly-D-Lysine (PDL) are two synthetic polymers with properties that could improve the performance of these interfaces. In my masters’ research, my objective is to determine whether these synthetic polymers could promote the formation of presynaptic elements on artificial surfaces in vivo making intraneural implants more biocompatible and long-lasting. Methods: In the coated microsphere in vivo experiment, a nerve crush injury model in the rat was used for the study. PDL-coated, DND-coated and uncoated beads were injected into the rat sciatic nerve at the crush site and 5 mm distal to the crush site. The nerves were then harvested after 4, 6 and 8 weeks and stained for neurofilament and synaptophysin. Synaptophysin puncta were then counted on the bead surface for each group. Additionally, in a proof-of-concept experiment, two uncoated electrodes were implanted into the rat sciatic nerve. Nerve recordings were then performed every week, and the threshold nerve potential in the sciatic nerve was measured by only varying the pulse duration of the stimulation. Results: The coated microsphere in vivo experiment demonstrated that DND-coated microspheres had a significantly higher number of synaptophysin puncta around their surface from 4 to 8 weeks compared to uncoated beads. At 4 weeks, the DND condition also showed a significantly higher number of synaptophysin puncta around its microbeads vs. the PDL condition for the distal site. In the uncoated electrode in vivo experiment, the results showed that the two implants could stimulate and record threshold nerve potentials in the rat sciatic nerve for one week and two weeks respectively before being non-functional. Conclusion: Our study showed for the first time that DND has a stable synapse-promoting property that is superior to PDL in vivo and that our electrode design can be used to assess the long-term signal stability of peripheral nerve implants.