Combinatorial techniques, which overcome limitations of actual models of material research permitting to effectively address this large amount of variables, are utilized in this work to prepare combinatorial libraries of the blend of the biodegradable polymers Poly(e-caprolactone) and Poly(lactic acid). These libraries present continuous composition and temperature gradients in an orthogonal fashion that permit to obtain multiple surface morphologies with controllable microstructures due to the blends low critical solution phase behavior (LCST). The goal of this study is to investigate the effect of surface morphology (surface chemical patterning and surface topography) on cell behavior. The varied surface topography of the libraries is used as a valuable tool that permits to assay the interaction between MC3T3-E1 cells and hundreds of different values of critical surface properties, namely, surface roughness and microstructure size. The outcome of this tool is a rapid screening of the effect of surface topography on cell behavior that is orders of magnitude faster than the standard 1-sample for 1 measurement techniques. The results obtained show that cells are very sensitive to surface topography, and that the final effect of surface properties on cell function is intimately related with the stage of the cell developmental process. Meaning that, for example, areas with optimal characteristics to elicit enhancement of cell attachment is not necessarily the same that promotes cell proliferation. This study imparts an improved understanding of an often neglected factor in biomaterials performance: surface morphology (particularly surface topography). The results provide a new insight into the importance of taking into consideration both chemistry and physical surface features for superior biomaterial design.

Surgical repair of torn rotator cuff tendons have a high rate of failure and does not address the underlying pathophysiology. Tissue engineering strategies, employing the use of multipotent progenitor cells or growth factors, represent potential therapies to improve the outcome of rotator cuff surgery. The use of glycosaminoglycan-based biomaterials in these therapies may enhance the effectiveness of cell and growth factor delivery techniques. Furthermore, understanding the cellular and molecular mediators in tendon overuse can help elucidate the causes of tendon degeneration. Thus the overall goals of this dissertation were to 1) develop heparin-based biomaterials to enhance cell pre-culture and maintain growth factor bioactivity and 2) characterize the histological and enzymatic changes in a supraspinatus tendon overuse model. To investigate the use of heparin in enhancing dynamic signaling, mesenchymal stem cells (MSCs) were encapsulated in heparin-containing hydrogels and evaluated for differentiation markers when cocultured with a small population of differentiated cells. To probe the effect of sulfation of heparin on the interactions with protein, selectively desulfated heparin species were synthesized and evaluated for their ability to bind and protect proteins. Finally, to develop a tendon overuse model that can become a test bed for testing future targeted therapeutics, an animal model was evaluated for tissue damage and protease activity. Together these studies represent a multi-pronged approach to understanding how tendon tissues become degenerative and for developing technologies to improve the biological fixation of tendon to bone in order to reduce the need for revision surgeries.

A queimadura de 3º grau é um dos maiores traumas a que um ser humano pode ser submetido e em geral são as lesões mais frequentes na população mundial, tratando-se de um importante problema de saúde pública. Pensando neste problema, observou-se que a utilização da membrana amniótica (MA) pode ser o melhor tratamento que esses pacientes possam receber. Ela atua em benefício da epitelização, facilita a migração e a adesão das células epiteliais basais, a matriz estromal possui inibidores de proteases que previne a apoptose e restaura o fenótipo epitelial, antibacteriana do córion e âmnion, além de proteger a ferida e atuar na redução da dor. A descelularização, trata-se da retirada de todas as células e núcleo da MA utilizada, como forma de se evitar qualquer tipo de histoincompatibilidade. O presente trabalho teve como objetivo avaliar a eficácia do uso da membrana amniótica descelularizada (MAD) no tratamento de queimaduras de 3o grau em humanos, testada em ratos da linhagem Wistar. Trata-se de uma pesquisa descritiva, a qual investiga fundamentalmente a especificação e a disposição de dados acerca dos resultados obtidos a partir de um processo regenerativo de queimaduras utilizando-se a MAD. A amostra foi constituída de constituída por 20 ratos machos adultos da linhagem Wistar, dividida em 2 grupos (n=10): grupo controle (GC)–sem MA e grupo transplantado–com MA (TMAD). Os dois grupos foram submetidos a um processo padronizado de queimadura térmica, sendo retiradas duas amostras de tecido para análise, no 14º dia e no 30º dia. As amostras foram analisadas com o emprego de técnicas anatomopatológicas onde foram montadas em lâminas fragmentos de MA corados com solução Hematoxilina Eosina (HE) e, para análise histomorfométrica,as lâminas foram coradas com picro sirius red sob luz polarizada para verificação da descelularização e quantificação de colágeno do tipo I, II e III. As imagens foram quantificadas utilizando o programa Image Pro Plus®, versão 5.0 para Windows®. Os dados estatísticos foram analisados utilizando o programa computacional SSPS v.21.0. Observou-se um aumento significativo da quantificação de colágenos do tipo III no 14º dia no grupo TMAD em relação ao GC, do colágeno do tipo II no 14º e 30º dia comparado ao GC e do colágeno tipo I no 14º e 30º dia, mostrando que a utilização da MA foi eficaz no grupo TMAD. Concluiu-se que a MAD, aplicada topicamente, demonstrou eficácia no processo cicatricial em queimaduras de 3º grau. Esperava-se que a mesma promovesse a aceleração de cicatrização dos ferimentos, mas, o que se observou ao término do trabalho é que a MAD não só promoveu a cicatrização como foi mais eficiente no processo de regeneração dos tecidos lesados. / 3rd degree of burns are one of the greatest injury that a human being can be submitted and generally are the most frequent injuries in the world population, in the case of a major public health problem. Thinking this problem, it has been observed that the use of amniotic membrane (AM) may be the best treatment these patients can receive. It acts on behalf of epithelialization, facilitates migration and adhesion of basal epithelial cells, stromal matrix has protease inhibitors which prevent apoptosis and restores epithelial, antibacterial phenotype of the chorion and amnion, and protect the wound and act on reducing pain. The decellularization, it is the withdrawal of all cells and core AM used as a way to avoid any kind of histoincompatibility. This study aimed to evaluate the efficacy of the decellularized amniotic membrane (DAM) in the treatment of third degree burns in humans, tested in Wistar rats. It is a descriptive research which investigates mainly the specification and the arrangement of data on the results obtained from a regenerative process burns using DAM. The sample consisted of consisted of 20 Wistar adult male rats, divided into 2 groups (n = 10): control group (CG) - without MA, and group transplanted - with AM (TDAM). The two groups were submitted to a standard process of thermal burn, two samples being removed tissue for analysis on day 14 and day 30. Samples were analyzed with the use of pathological techniques which were mounted on slides DNA fragments stained with solution hematoxylin eosin (HE) for histomorphometric analysis, the slides were stained with picrosirius red under polarized light for verification of decellularization and quantifying collagen type I, II and III. Images were quantified using Image Pro Plus ®, version 5.0 for Windows. Statistical data were analyzed using the computer program SPSS v.21.0. There was a significant increase in type III collagen quantification on the 14th day in TDAM group compared to CG, type II collagen in the 14th and 30th day compared to the CG and type I collagen in the 14th and 30th day, showing that the use of MA were effective in TDAM group. It was concluded that the DAM, applied topically, has shown efficacy in the healing process in 3rd degree burns. It was expected that it would promote accelerated wound healing but which was observed at the end of work that MAD is not only promoted as the healing was more efficient the regeneration process of the damaged tissues.

CNPQ / O presente trabalho tem como objetivo o desenvolvimento de um biosensor, utilizando redes de Bragg em fibras óticas, capaz de mensurar deformações em tecido ósseo. Inicialmente foram realizados testes de esterilização e desinfecção com sensores, após o processo completo, os sensores foram testados quanto a sua sensibilidade. Com isso os métodos de esterilização e desinfecção autoclave, óxido etileno, hipoclorito de sódio, cloramina T e ácido peracético, foram eficazes para eliminação e ou inativação dos microrganismos, sem que haja alterações no comportamento das FBGs. Com isso definem-se métodos aplicáveis para possíveis estudos, onde fibras óticas foram implantadas em animais in vivo para estudos de comportamento metabólico com a presença de materiais inorgânicos. Na continuidade dos trabalhos foram realizados testes de compatibilidade de fibras óticas inseridas no tecido subcutâneo de ratos. Baseado na análise descritiva dos cortes histológicos pode-se concluir que as fibras ópticas sem revestimento e com revestimento foram biocompatíveis quando implantadas em subcutâneo de ratos. Sendo assim, foi possível contemplar o desenvolvimento de um sensor baseado em redes de Bragg em fibras óticas para implantação direta no tecido ósseo. Posteriormente foram realizados testes-piloto para o desenvolvimento da metodologia de construção e implante no desenvolvimento de biosensores. Com todo o trabalho feito até à data, o desenvolvimento e construção de um biosensor que foi implantado in vivo foi possível. O biossensor foi fixado na região mandibular de um bovino quatro meses de idade. Uma semana após a implantação, os animais receberam diferentes tipos de alimentos e foi monitorizada durante a ingestão de alimentos. Com as deformações medidas foi possível a análise dos diferentes padrões mastigatórios e diferentes magnitudes de força e frequência mastigatória que o animal desempenhada durante o processo de mastigação. / This study aims to develop a biosensor using Bragg gratings in optical fibers, capable of measuring deformation in bone tissue for this took a lot of work. Initially sterilization and disinfection with sensors tests were performed after the entire process, the sensors were tested for their sensitivity. In the same study methods of sterilization and disinfection have been tested to assess its efficiency. Thus methods of sterilization and disinfection autoclave, ethylene oxide, sodium hypochlorite, chloramine T and peracetic acid were effective for removing or inactivating microorganisms and, without changes in the behavior of the FBGs. With this set up methods applicable for possible studies, where optical fibers are implanted in in vivo animal studies metabolic behavior in the presence of inorganic materials. Continuing the work was carried out optical fiber compatibility tests inserted in the subcutaneous tissue of rats. Based on the descriptive analysis of histological sections can be concluded that the optical fiber uncoated and coated were biocompatible when implanted in rat subcutaneous tissue. Thus it is possible to contemplate the development of a sensor based on Bragg gratings in optical fibers for direct implantation into the bone tissue. Following on from the work were carried out pilot tests for development of construction methodology and implant in biosensor development. With all the work done to date, the development and construction of a biosensor that was implanted in vivo was possible. The biosensor was fixed in the mandibular region of a bull four months old. A week after implantation, the animal received different types of food and was monitored during the intake of foods. With the deformations measured parse the different masticatory patterns and different magnitudes of force and masticatory frequency that the animal developed during the chewing process completing the objectives of this study.

Made available in DSpace on 2015-03-26T13:47:24Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2414435 bytes, checksum: e0ee8e2ec29d15623795df16ecdca852 (MD5) Previous issue date: 2014-07-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Periodontal disease is a major problem both in human and in veterinary medicine, because presents as a serious consequence of the loss of alveolar bone, besides the supporting tissues of the dental piece: cementum and periodontal ligament. The treatment of this disease is the manual removal of plaque associated with the use of different antimicrobials to prevent systemic consequences that disease can cause. However, the conventional treatment prevents the formation of periodontal tissue support, occurring only proliferation of cells of the junctional epithelium and gingival connective tissue fibroblasts. Bone defects resulting from the need to fill condition replacements for bone regeneration to occur at the appropriate location. The use of biocompatible and osteoconductive biomaterials, preferably resorbable, serves as support for bone formation and are gradually reabsorbed by the body without any adverse reaction. The aim of this study was to characterize two membranes composed of synthetic hydroxyapatite (HAP 91 PLUS ®) and polycaprolactone (PCL) in different concentrations (75HAP/25PCL and 60HAP/40PCL) and it evaluation of the use in bone defects in the alveolar bone of the jaw dogs. The analysis of the membrane by scanning electron microscopy revealed complex surface in the membrane 75HAP/25PCL and less complex in the membrane 60HAP/40PCL. Both showed high crystallinity, evaluated by x-ray diffraction, and the presence of hydroxyapatite crystals adhered to the membrane, which improve their complex topography. The average pore diameters were 15.7 (± 8.1)μm and 10.4 (± 6.3) μm without, however, show the presence of macropores or interconnection between the existing pores. The experimental groups were divided into control and treated, containing six animals each. Clinical evaluations and radiographs were performed two weeks after the implantation of the membrane and 60 days after operation. Histology and microtomography scan was performed at 60 days postoperatively. Were not clinically observed pain and purulent discharge, and the clinical insertion level showed no statistical difference between the groups. In treated animals, 50% of the animals showed dehiscence and 83% had membrane exposure. Radiographic images show alveolar bone radiopacity equal in both groups. The computed microtomography revealed that the control group showed, with statistical difference, the greater ratio of bone volume over the total volume of the defect in relation to the treated group, however in all animals, the furcation area was not filled by new alveolar bone. Histology revealed the presence of extensive inflammatory infiltration and connective tissue in both groups. New bone was only observed in the apical border of the defect in both the control and the treated group. / A doença periodontal é um importante problema tanto na medicina humana quanto na veterinária, pois apresenta como grave consequência a perda dos tecidos de sustentação da peça dental, sendo que o tratamento convencional retirada mecânica da placa - não permite a formação dos tecidos periodontais de sustentação. Os defeitos ósseos resultantes da afecção necessitam de preenchimento com substitutos para que ocorra a regeneração óssea apropriada no local. A utilização de biomateriais biocompatíveis e osteocondutores, preferencialmente reabsorvíveis, serve como suporte para formação óssea e são reabsorvidos gradativamente pelo organismo, sem qualquer reação adversa. O objetivo desse trabalho foi caracterizar duas membranas compostas por hidroxiapatita sintética (HAP 91 PLUS ®) e policaprolactona (PCL) em diferentes concentrações (75HAP/25PCL e 60HAP/40PCL) e avaliação da utilização da segunda em defeitos periodontais na mandíbula de cães. A análise da membrana pela microscopia eletrônica de varredura revelou superfície complexa na membrana 75HAP/25PCL e menos complexa na membrana 60HAP/40PCL. Ambas apresentaram alta cristalinidade, avaliadas pela difração de raios-X e a presença de cristais de hidroxiapatita na superfície da membrana nos quais auxiliam a complexidade da topografia. Os diâmetros médios dos poros foram de 15,7(±8,1) μm e 10,4 (±6,3) μm sem, entretanto, evidenciar a presença de macroporos nem a interconexão entre os poros existentes. Para explicação in vivo, os grupos experimentais foram divididos em controle e tratado, contendo seis animais cada. A doença periodontal foi induzida em três estágios em todos os animais, e seis deles receberam tratamento com a técnica de regeneração tecidual guiada. As avaliações clínicas foram realizadas durante duas semanas após a implantação da membrana e após 60 dias do pós-operatório. Radiografias foram retiradas no pré operatório imediato e aos 60 dias após tratamento. A histologia e a tomografia microcomputadorizada foram realizadas aos 60 dias do pós-operatório. Clinicamente não foram observados dor e secreção purulenta, e não houve diferença estatística no nível clinico de inserção entre os grupos avaliados. Nos animais tratados, 50% apresentaram deiscência e 83% exposição de membrana. As imagens radiográficas demonstraram radiopacidade igual ao osso alveolar na região do defeito em ambos os grupos. A tomografia microcomputadorizada revelou que o grupo controle apresentou, com diferença estatística, maior relação de volume ósseo sobre o volume total do defeito em relação ao grupo tratado, no entanto em todos os animais, a região de furca não foi preenchida por novo osso alveolar. A histologia revelou presença de grande infiltrado inflamatório e presença de tecido conjuntivo em ambos os grupos. Osso novo apenas foi observado na borda apical do defeito tanto no grupo controle quanto no grupo tratado. Assim, o biomaterial utilizado não obteve sucesso quando utilizado em defeitos periodontais, sendo necessários assim, maiores estudos quanto a sua utilização em outras regiões do organismo.

Os equipamentos médicos, sob regime da Vigilância Sanitária, compreendem todos os equipamentos de uso em saúde com finalidade médica, odontológica, laboratorial ou fisioterápica, utilizados direta ou indiretamente para diagnóstico, terapia, reabilitação ou monitorização de seres humanos. A manutenção desses equipamentos é atribuída a área de engenharia clínica. A engenharia clínica atua no desenvolvimento, gerenciamento, manutenção, incorporação e capacitação dos equipamentos médicos-hospitalares (EMH) dentro do hospital, visando contribuir com a qualidade de vida dos pacientes. Para melhorar a gestão destes equipamentos, aliou-se a informática, por meio dos sistemas de informação à área de manutenção. Os softwares destinados à área hospitalar existentes, são na maioria comerciais, caros e estão voltados à gestão dos pacientes, gestão de leitos e prontuários. A existência dos sistemas de informatização com a finalidade de auxiliar na manutenção, pode reduzir o tempo e o custo destinado a esta atividade. O objetivo deste estudo é analisar e implementar um sistema de informação para gerenciamento de manutenção de EMH. Este sistema é 100% Web, multiplataforma, que usa um método de priorização para manutenções preventiva e corretivas. Foram criados formulários personalizados para cada EMH visando a padronização de como serão armazenadas as soluções de cada manutenção. Para o desenvolvimento do sistema, foi aplicada a Análise Orientada a Objetos, por meio de técnicas da engenharia de software. O código do sistema foi desenvolvido usando a linguagem de programação PHP, utilizando um framework frontend Bootstrap e um banco de dados PostgreSQL. Com o sistema pronto foram criados métodos de testes de software disponibilizando roteiros para testar a usabilidade do software. Comparando o SIGEMH e outros softwares disponíveis no mercado, foi constatado que a maioria contempla os aspectos gerenciais de EMH, enquanto que o SIGEMH tem como meta a priorização e a padronização de formulários referentes às manutenções. Algumas funcionalidades existentes no outros softwares não foram desenvolvidas, mas implementou-se novas funcionalidades, tais como: o armazenamento de normas técnicas e manuais para pesquisas on-line, a padronização dos formulários pós manutenção e o método de priorização que define as ordens dos equipamentos para a realização dos serviços. / The medical equipment, under sanitary vigilance rules, comprehend all equipment in use in the health field with medical, odontological, laboratorial or physiotherapeutic purpose, which are used either directly or indirectly for diagnosis, therapy, rehabilitation, or monitoring of human beings. The maintenance of these equipment is attributed to the clinical engineering area. The clinical engineering acts on development, management, maintenance and capacitation of medical-hospital equipment (MHE), within the hospital, aiming to contribute to the patients? quality of life. In order to improve the management of these equipment, it has allied with informatics, by means of information systems in the maintenance field. Most of the existing softwares for the hospital area are commercial, expensive and are meant to patients, beds and medical records managing. The existence of informatization systems with the aim of helping maintenance may cut down the time and the cost for this activity. The purpose of this study is to analyze and to implement an information system for managing the maintenance of MHE. This system is 100% Web, multiplatform, and uses a prioritizing method for preventive and corrective maintenance. Customized forms were created for each MHE, aiming the standardization of storage of solutions for each maintenance. For the developing of the system, the Object Oriented Analysis was applied, with techniques of software engineering. The system code was developed using PHP programming language, using a Bootstrap frontend framework and a PostgreSQL database. When the system was ready, software testing methods were created, providing itineraries to test the software?s usability. Comparing to SIGEMH and other softwares on the market, it was found that the majority regards the managing aspects of MHE, while SIGEMH aims the prioritization and standardization of forms relative to maintenance. Some functionalities present in other softwares were not developed, but new functionalities were implemented, such as: storage of technical rules and manuals for online searching, standardization of post-maintenance forms and the method which defines the order of equipment for service execution.

Une lésion au niveau d’un nerf périphérique peut provoquer la perte de fonction sensorielle et motrice, et dans le cas de neurotmésis, la régénération spontanée ne se produira pas. De plus, si l’espace entre les deux segments de nerf est trop important, une suture directe n’est pas possible et l’implantation d’une greffe est nécessaire afin de créer une liaison entre les deux segments de nerf. L’autogreffe de nerf est le « gold standard » pour des procédés de réparation nerveuse : une portion d’un nerf sein (qui est considéré comme un nerf moins important) est prise du même patient et implantée au site de la lésion. Cependant, il existe plusieurs désavantages avec ce procédé comme une deuxième chirurgie, la perte de fonction au site du don, la possibilité de développer un neurome sur ce même site, ainsi qu’un taux de réussite de 50% dans les cas où l’espace entre les deux segments de nerf est très important. Il reste donc, un besoin de trouver un procédé alternatif afin d’augmenter le taux de réussite et d’éliminer les désavantages de l’autogreffe. L’objectif de cette étude est d’avancer vers une solution alternative de l’autogreffon en utilisant des biomatériaux. Cette thèse se divise en trois parties. La première se focalise sur le développement d’un modèle de guide nerveux basé sur des nanofibres de fibroïne de soie. Ce matériau est composé d’une organisation complexe qui inclut deux surfaces de nanofibres alignées avec une couche de nanofibres aléatoires à l’intérieur afin d’améliorer des propriétés mécaniques du matériau sans la perte d’orientation des fibres pour la régénération nerveuse. Le matériau est ensuite manipulé pour fabriquer un tube, multi-canaux avec une « enveloppe » supplémentaire afin de faciliter le procédé d’implantation chirurgicale. Ce guide nerveux a été soumis pour l’obtention d’un brevet européen le 12 juillet 2017 et cela est le sujet d’un deuxième article qui a été soumis pour publication. La deuxième partie de cette étude explore des possibilités d’une fonctionnalisation du matériau afin d’améliorer son efficacité pour la régénération nerveuse. Cette étude explore la fonctionnalisation de la fibroïne de soie avec une deuxième protéine, plusieurs facteurs de croissance, et des nanoparticules. Chacune de ces fonctionnalisations donne une possibilité d’ajouter des propriétés favorables à la fibroïne de soie, un matériau naturel et biocompatible. La troisième partie de cette étude examine l’efficacité d’un guide nerveux composé de la fibroïne de soie fonctionnalisée avec des facteurs de croissance pour la régénération nerveuse périphérique en comparaison avec un guide nerveux composé de la fibroïne de soie sans aucune fonctionnalisation et une suture direct (qui simule une autogreffe). Trois techniques d’évaluation différentes de la régénération nerveuse ont été réalisées afin d’obtenir une analyse plus complète. Il y a de nombreux mécanismes impliqués dans la régénération nerveuse, il est donc nécessaire d’étudier différents paramètres pour analyser l’efficacité de régénération. Les résultats d’analyses histologiques, d’électromyographie, et de capture de mouvement, ont été considérées ensemble afin d’arriver à une conclusion sur la réussite d’une régénération nerveuse pendant cette étude. Pour conclure cette étude, les guides nerveux fonctionnalisés avec une combinaison de facteurs de croissance démontrent une meilleure régénération nerveuse et une récupération de fonction supérieure. / Injury to a peripheral nerve can cause loss of sensory and motor function, and if the injury is very severe where the nerve undergoes neurotmesis, unassisted nerve regeneration may not occur. In this case, where the gap between nerve segments is too large to carry out a direct end to end suture, a graft is sutured to bridge the gap between sectioned nerve segments. The autologous nerve graft, where a portion of a less important nerve from the same patient is removed and grafted between nerve segments, continues to be the gold standard procedure for nerve repair. However, there are several drawbacks of this technique including a second surgical procedure, loss of function at the donor site, possibility of developing a painful neuroma at the donor site, and the 50% success rate of autografts used in large gaps. There is therefore a need for a tissue engineered nerve graft that can replace the autograft, and this study aims to advance toward an effective autograft alternative. This PhD is presented as a three part study consisting first of the development of a novel nerve guidance conduit based on a tri-layered silk fibroin nanofiber material comprised of a complex organization including two aligned fiber surfaces and a randomly deposited fiber interior to improve the mechanical properties of the material while not compromising the guidance capabilities of aligned nanofibers for nerve regeneration. The material is then used to fabricate a multi-channeled tube with an additional “jacket layer” in order to facilitate surgical implantation. This NGC has been submitted to be patented on July 12, 2017 and is the subject of the second article submitted for review for publication. The second part of this study explores the different possibilities of the functionalization of the material in order to improve the effectiveness for nerve regeneration. This study explores functionalizing the silk fibroin with a second protein, several growth factors, and nanoparticles that all have potential to add favorable properties to the natural biocompatible silk fibroin material. The final part of this study tests the effectiveness of growth factor-embedded silk fibroin NGCs for peripheral nerve regeneration in comparison with non-functionalized silk fibroin devices and a direct suture to simulate results obtained with an autograft. Three different techniques for the evaluation of nerve regeneration were used in order to produce a more comprehensive analysis. As there are many mechanisms involved in nerve regeneration, only one or two analysis techniques cannot paint a complete picture of the success of nerve regeneration. Therefore, histological analyses, electromyography analyses, and motion capture analyses were carried out and considered together in order to make a conclusion on the level of nerve regeneration success during this study. The conclusions from this study were that a NGC functionalized with a combination of growth factors appeared to exhibit the most successful nerve regeneration and functional recovery.

La lésion de nerfs périphériques peut engendrer des déficits moteurs et/ou sensoriels permanents. En dépit des progrès techniques réalisés au cours de ces 25 dernières années, une récupération complète suite à ces lésions n’est pas encore possible aujourd'hui. L’autogreffe nerveuse, toujours considérée comme le standard clinique, est la seule technique capable d’offrir les meilleurs résultats en termes de récupération fonctionnelle. Cependant, la survenue de complications post-opératoires lors d’autogreffes d’un nerf et la quantité limitée de nerfs disponibles conduisent à mettre au point d’autres stratégies alternatives. Dans ce contexte, la mise au point de biomatériaux pour substituts nerveux devient une nécessité clinique. Malgré les efforts de la recherche, ces prothèses ne permettent toujours pas une régénération du nerf à la hauteur de l’autogreffe. Le biomatériau utilisé doit notamment présenter des propriétés physiques et chimiques proches de celui du nerf natif. La soie, aux propriétés mécaniques uniques, représente une bonne alternative pour mettre au point ce type de prothèses. En effet, la protéine de soie déjà utilisée dans le domaine biomédical est biocompatible. Les modifications chimiques de cette protéine améliore et favorise l’adhérence et la croissance cellulaires par l’incorporation de facteurs de croissance ou d’autres molécules d'intérêt. Ce travail de thèse propose de développer un nouveau type de biomatériau à base de soie fonctionnalisée par deux facteurs de croissance : le Nerve Growth Factor (NGF) et le Ciliary NeuroTrophic Factor (CNTF). Étant donné l’architecture complexe qui compose la structure nerveuse, une matrice supportant la repousse des tissus de façon orientée semble primordiale. Nous démontrons, dans un premier temps, le pouvoir de ces nanofibres alignées (produites par electrospinning) à orienter la régénération tissulaire de différents organes par culture d’explants. Les nanofibres de soie alignées, biocompatibles sont bio-activées par ajout de NGF spécifique de la régénération nerveuse. Cette matrice créée présente un gradient de concentration en NGF qui permet d’orienter la repousse axonale en stimulant la croissance axonale dans une seule direction. Afin d’optimiser la croissance de deux populations cellulaires, nous avons incorporé du CNTF pour produire des nanofibres bifonctionnalisées. Ces nanofibres bifonctionnalisées ont conduit à une longueur des neurites 3 fois plus grande à leurs contacts, stimulant la croissance des cellules gliales. Ainsi, nous avons produit des conduits nerveux à base de soie biofonctionnalisée pour implantation chez le rat. Les analyses physico-chimiques et les propriétés mécaniques démontrent le caractère biomimétique de nos tubes de guidage. Les premières études de la locomotion et l’observation de coupes du nerf sciatique de rat, suite à l’implantation de nos conduits donnent des résultats très prometteurs. L’ensemble de ces travaux démontre l’efficacité de nos guides nerveux à base de soie et les présente comme une alternative prometteuse à l’autogreffe nerveuse pratiquée en clinique. / Peripheral nerve injury causes sensory and/or motor functions deficits. Despite technological advances over the past 25 years, a complete recovery from these injuries remains unsatisfactory today. The autograft still considered the "gold standard" in clinical practice. This is the only technique able to offer complete functional recovery. However, the occurrence of postoperative complications in autologous nerve and the limited amount of available nerves lead to develop alternatives strategy.In this context, development of nerve graft substitutes becomes by far a clinical necessity. Despite research efforts, these artificial prostheses design based on biomaterial doesn’t allow nerve regeneration as found in autograft nerve procedures. The biomaterial used must have the physical and chemical properties similar to that of the native nerve. Silk, well known for its unique mechanical properties, proposes a good alternative to develop these prostheses. Indeed, the silk protein is commonly used in the biomedical field and regenerative medicine. This protein biocompatibility may be improved through chemical modifications to promote adhesion and cell growth by the incorporation of growth factors or other molecules of interest. Therefore, this thesis proposes to develop a new type of functionalized silk biomaterial based on two growth factors : Nerve Growth Factor (NGF) and Ciliary NeuroTrophic Factor (CNTF). Given the complex architecture that consists of nerve structure, a matrix which is able to support and manage the outgrowth of tissue becomes essential. We demonstrate the power of these aligned nanofibers (produced by electrospinning) to guide and manage tissue regeneration from different organ explants culture. Aligned silk nanofibers, were biocompatible and bio-activated by adding NGF involved for nerve regeneration. This matrix has been created with a concentration gradient of NGF to guide neuritis outgrowth in only one direction. The presence of this gradient demonstrated a better axonal growth in one direction versus the uniform concentration conditions. Nerve cells consist essentially of two cell populations which are neurons and Schwann cells. To optimize the culture and growth of these two populations, in addition to NGF, we incorporated CNTF to produce bifunctionalized nanofibers. These biofunctionalised nanofibers led to a length 3 times larger on contact with neurites. The glial cells growth, alignment and migration were stimulated by CNTF. Thus, we produced bi-functionalized nerve guidance conduits for rat implantation. The physico-chemical analyzes demonstrate the biomimetic of our guide tubes. Early studies of locomotion and observing histological sections of rat sciatic nerve, following the implementation of our conduits gave very promising results.These studies demonstrate the relevance of our nervous guides’ silk-based developed as an effective alternative to nerve autograft performed in the clinic.

Researches on alloy elements such as niobium and silicon have been added to titanium alloys as an alternative for new materials to be used in orthopedic implants, once they present biocompatibility and favor reductions in the elasticity modulus. Therefore, the objective of this study was to investigate the phase transformations, mechanical properties and corrosion resistance of the Ti-Nb and Ti-Nb-Si alloys for biomedical applications. The results showed that Si addition is effective to stabilizing the £] phase, which also acts as a ç suppressor, and to produce grain refinement. In all samples, the hardness values increased with the increasing of Si content, while the elastic modulus was reduced in the Ti-15Nb-xSi and Ti-35Nb-xSi alloys; however, for Ti-40Nb-xSi samples increased with the increasing of Si content. Lower elastic moduli were observed in water-quenched Ti-35Nb-xSi alloys, for which tensile tests and corrosion resistance were performed. The increase in Si concentration also produced an increase in the alloys mechanical strength. Ti-35Nb-0,55Si alloy, however, showed brittle mechanical behavior which was related to the presence of silicides, referred as (Ti,Nb)3Si, at the grain boundaries. Corrosion tests showed that the alloys exhibit high resistance in the studied environment. The results suggest that the Ti-35Nb-0,35Si alloys presents the best combination of properties: good mechanical strength, low elastic modulus and excellent corrosion resistance. / Elementos de liga como niobio e silicio tem sido adicionados a ligas de titanio como uma alternativa na busca de novos materiais para implantes ortopedicos, visto que apresentam biocompatibilidade e favorecem a reducao do modulo de elasticidade. Dessa forma, o objetivo deste trabalho foi investigar as transformacoes de fases, propriedades mecanicas e de resistencia a corrosao de ligas dos sistemas Ti-Nb e Ti-Nb-Si para aplicacoes biomedicas. Os resultados mostraram que a adicao de Si foi efetiva na estabilizacao da fase £], o qual tambem atua como supressor de ç, alem de produzir refinamento de grao. Em todas as amostras estudadas, os valores de dureza se elevaram com o aumento do teor de Si, enquanto que o modulo de elasticidade sofreu uma reducao nas ligas dos grupos Ti-15Nb-xSi e Ti-35Nb-xSi; no entanto, para as amostras Ti-40Nb-xSi foi verificado um aumento de seus valores com a elevacao do teor de Si. Menores modulos de elasticidade foram observados nas ligas Ti-35Nb-xSi resfriadas em agua, para as quais foram realizados ensaios de tracao e de resistencia a corrosao. Foi observado que o aumento da concentracao de Si tambem produziu um aumento na resistencia mecanica das ligas, entretanto, a liga com 0,55% de Si, apresentou comportamento mecanico fragil atribuido a presenca de silicetos, indicados como (Ti,Nb)3Si nos contornos de graos. Os ensaios de corrosao mostraram que as ligas apresentaram uma alta resistencia no meio estudado. Os resultados obtidos sugerem que a liga Ti-35Nb-0,35Si apresenta a melhor combinacao de propriedades: boa resistencia mecanica, baixo modulo de elasticidade e excelente resistencia a corrosao.

Indiana University-Purdue University Indianapolis (IUPUI) / Hydrogels prepared from photopolymerization have been widely used in many biomedical applications. Ultraviolet (200-400 nm) or visible (400-800 nm) light can interact with light-sensitive compounds called photoinitiators to form radical species that trigger photopolylmerization. Since UV light has potential to cause cell damage, visible light-mediated photopolymerization has attracted much attention. The conventional method to fabricate hydrogels under visible light exposure requires usage of co-initiator triethanolamine (TEA) at high concentration (∼200 mM), which reduces cell viability. Therefore, the first objective of this thesis was to develop a new method to form poly(ethylene glycol)-diacrylate (PEGDA) hydrogel without using TEA. Specifically, thiol-containing molecules (e.g. dithiothreitol or cysteine-containing peptides) were used to replace TEA as both co-initiator and crosslinker. Co-monomer 1-vinyl-2-pyrrolidinone (NVP) was used to accelerate gelation kinetics. The gelation rate could be tuned by changing the concentration of eosinY or NVP. Variation of thiol concentration affected degradation rate of hydrogels. Many bioactive motifs have been immobilized into hydrogels to enhance cell attachment and adhesion in previous studies. In this thesis, pendant peptide RGDS was incorporated via two methods with high incorporation efficiency. The stiffness of hydrogels decreased when incorporating RGDS. The second objective of this thesis was to fabricate hydrogels using poly(ethylene glycol)-tetra-acrylate (PEG4A) macromer instead of PEGDA via the same step-and-chain-growth mixed mode mechanism. Formation of hydrogels using PEGDA in this thesis required high concentration of macromer (∼10 wt.%). Since PEG4A had two more functional acrylate groups than PEGDA, hydrogels could be fabricated using lower concentration of PEG4A (∼4 wt.%). The effects of NVP concentration and thiol content on hydrogel properties were similar to those on PEGDA hydrogels. In addition, the functionality and chemistry of thiol could also affect hydrogel properties.