Orientador: Carlos Roberto Grandini / Banca: Sandra Giacomin Schnider / Banca: Eduardo Carlos Bianchi / O Programa de Pós-Graduação em Ciência e Tecnologia de Materiais, PosMat, tem caráter institucional e integra as atividades de pesquisa em materiais de diversos campi da Unesp / Resumo: A utilização de biomateriais do reparo do tecido ósseo tem revolucionado a Ortopedia e a Odontologia atuais. Um metal estudado para estas aplicações é o titânio e suas ligas, pois possuem um excelente grau de biocompatibilidade com o organismo humano, uma excelente resistência à corrosão em meio biológico e uma ótima relação resistência mecânica/densidade. Estas propriedades são fortemente influenciadas pela presença de lementos intersticiais e as medidas de atrito interno têm sido muito utilizadas para verificar o comportamento destes elementos em materiais. O objetivo neste trabalho foi estudar o efeito de elementos intersticiais nas propriedades da liga Ti-10Mo (% em peso), por meio de medidas de difração de raios X, microscopia eletrônica e varredura, testes de citotoxidade, corrosão e espectroscopia mecânica. As amostras foram submetidas a tratamentos térmicos de recozimento e dopagem com oxigênio. Os testes de biocompatibilidade mostraram que não houve nenhuma resposta indireta. Os extratos dos materiais não interferiram no desenvolvimento, crescimento e proliferação das células. Foi detectada uma melhora na taxa de corrosão, juntamente com um aumento do potencial de corrosão, com a dopagem do material, características estas interessantes para um biomaterial. Os resultados de espectroscopia mecânica mostraram a presença de uma estrutura de relaxação após a dopagem com oxigênio em torno de átomos dos elementos que compõem a liga. Observou-se que a adição de oxigênio intersticial causou um aumento no módulo de elasticidade, o que é ruim quando se trata de um biomaterial para implantes ortopédicos. / Abstract: The use of the biomaterials in the repair of the bone tissue has been revolutionizing the Orthopedics and the Odontology. A metal very studied for these applications is the titanium and its alloys, because they possess an excellent corrosion resistance in a biological media and a good relationship mechanical strength/density. These properties are strongly influenced by the presence of interstitial elements and the internal friction measurements have been a lot used to verify the behavior of these interstitial elements in materials. The objective of this work was to study the effect of interstitial elements in the properties of the Ti-10Mo (%in weight) alloy, through X-ray diffraction, scanning electron microscopy, cytotoxicity tests, corrosion and mechanical spectroscopy. The samples were submitted to heat treatments of annealing and oxygen doping. The biocompatibility tests showed that there was not any indirect cytotoxic answer. The extracts of the materials didn't interfere in the development, growth and proliferation of the cells. An increase in the corrosion rate was detected, with an increase of the corrosion potential, with the oxygen doping, interesting for applications as biomaterial. The mechanical spectroscopy results showed the presence of a relaxation structure after the oxygen doping that was attributed mainly to the stress-induced ordering of the oxygen atoms around the atoms that compose the alloy. It was observed an increase in the elasticity with the oxygen doping, what is bad when it is a biomaterial. / Mestre

Orientador: Carlos Roberto Grandini / Banca: Paulo Afonso Silveira Francisconi / Banca: Ana Paula Rosifini Alves Claro / O Programa de Pós-Graduação em Ciência e Tecnologia de Materiais, PosMat, tem caráter institucional e integra as atividades de pesquisa em materiais de diversos campi da Unesp / Resumo: O crescente aumenta na expectativa de vida da população tem resultado numa igual demanda e procura por biomateriais metálicos. Biomateriais metálicos são utilizados na substituição ou recuperação de ombros, joelhos, quadril e cotovelos. As ligas a base de Ti são materiais promissores para tais fins, pois apresentam uma excelente biocompatibilidade, ótima resistência à corrosão em meio biológico. Essas características são grandemente influenciadas pela presença de elementos substitucionais e oxigênio intersticial e a técnica do atrito interno constitui um aliado bastante importante na análise dos resultados deste trabalho. Dessa forma, o objetivo deste trabalho é analisar a influência do exigênio intersticial nas propriedades das ligas Ti-8Ta e Ti-16Ta (% em peso). As medidas de atrito interno foram utilizadas para obtenção do módulo de elasticidade dinâmico das ligas estudadas, pois esta grandeza está experimentalmente associada ao amortecimento das vibrações livres do sistema, a dissipação de energia elástica e a frequencia de oscilação do sistema. Para tais medidas empregou-se um Pêndulo de Torção com frequencia de oscilação de aproximadamente 30Hz e temperatura na faixa de 230 a 350K. As ligas estudadas passaram por processos de tratamento térmico e dopagens com oxigênio e foram caracterizadas por medidas de densidade, difração de raios X e posterior refinamento de Rietveld, microdureza, módulo de elasticidade e testes de citotoxicidade in vitro. Os testes iniciais de biocompatibilidade mostram que não houve nenhuma resposta citotóxica, originando resposta celular satisfatória. As medidas de microdureza apresentam valores menores que os encontrados na literatura e as medidas de raios X mostram picos característicos da fase 'alfa' do titânio. Na análise de Rietveld verificou-se a presença de estrutura martensítica 'alfa' e fase 'beta' para ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The accelerating increase in life expectancy of the population has resulted in greater demand and more searches for metallic biomaterials. Metallic are extensively used in the replacement or restoration fo shoulders, knees, hips and elbows. Ti-based alloys are promising materials for such purposes, because they present excellent biocompatibility, a great mechanical strengh/density relation and a high corrosion resistance in a biological medium. These characteristics are greatly influenced by the presence of substitutional elements and interstitial oxygen. The internal friction technique is a very important ally in the analysis of the results of this work. Thus, the purpose of this work is to analyze the influence of insterstitial oxygen on the properties of Ti-8Ta and Ti-16Ta alloys (wt%). Internal friction measurements were used to obtain the dynamic elasticity modulus of the alloys studied, because this quantity is experimentally associated with the damping of free vibration of the sytem, which is related to the elastic energy dissipation and the oscillation frequency of the system. For such measurements, a torsion pendulum was employed, with an oscillation frequency of approximately 30 Hz and temperature in the range of 230-350 K. The alloys underwent heat treatment and oxygen doping processes, and were characterized by density, x-ray diffraction and subsequent refinement of Rietveld, microhardness and elasticity modulus measurements, and in vitro cytotoxicity tests. The initial biocompatibility tests showed that there was no cytotoxic answer, giving origin to a satisfactory cellular response. Microhardness measurements had values less than found in the literature, and x-rays showed characteristic peak of the titanium 'alfa' phase. In the Rietveld analysis, observed were the presence of martensitic 'alfa' structure and a 'beta' phase for all sample and conditions studied. The ... (Complete abstract click electronic access below) / Mestre

O objetivo desse estudo foi avaliar o comprometimento de pequenas vias aéreas na Fibrose Pulmonar Idiopática (FPI) analisando curvas Pressão- Volume (PV) do sistema respiratório. Coletamos curvas PV de doze pacientes antes da biopsia pulmonar, que confirmou FPI em sete pacientes e Pneumonite de Hipersensibilidade em cinco. Todas as curvas foram ajustadas com modelo sigmóide, V = a + b / (1 + e -(P-c/d)), e exponencial V = A - B . e -k.P (aplicado apenas à parte superior). O modelo exponencial, apesar do bom ajuste à parte superior, não representou a parte inicial da curva, gerando parâmetros sem significado. O modelo sigmóide ajustou bem toda a curva e gerou parâmetros com significado fisiológico, que sugerem a presença de colapso de pequenas vias aéreas na FPI. / The objective of this study was to evaluate small airways compromise in Idiopathic Pulmonary Fibrosis (IPF) using pressure-volume (PV) curves of the respiratory system. We collected PV curves from twelve patients before lung biopsy, which confirmed IPF in seven patients and Hipersensitivity Pneumonia in five. All curves were fitted with a sigmoid model, V = a + b / (1 + e -(P-c/d)), and an exponential model, V = A - B . e -k.P (applied only to the superior part of the curve). The exponential model, despite having a good fit to the superior part of the curve, did not represent the initial part, and yielded parameters with no physiological meaning. The sigmoid model had a good fit to the entire curve and yielded parameters with physiological meaning, suggesting the presence of small airways collapse in IPF.

Doenças pulmonares intersticiais fibrosantes são doenças que afetam homens, mulheres e crianças, tem prognóstico ruim e os pacientes possuem sobrevida estimada entre 3 e 5 anos após a confirmação diagnóstica, sobretudo os portadores de Fibrose pulmonar idiopática. Estudos recentes demonstram a capacidade das células-tronco em se diferenciar em diferentes linhagens celulares e tecidos, como já comprovado em órgãos como coração, fígado, trato gastrointestinal, sistema nervoso e pulmão. Os objetivos deste trabalho foram os de estabelecer a espécie suína como modelo experimental e utilizar a terapia celular experimentalmente como possibilidade de estudo para tratamento das doenças pulmonares intersticiais fibrosantes. Para estabelecer o modelo experimental e induzir a doença nos dois grupos de animais estudados (grupos tratado e controle) foi utilizado sulfato de bleomicina pela via intratraqueal em procedimento único. Após a instalação da doença, os animais dos grupos tratado e controle foram submetidos a tomografia computadorizada de alta resolução (TCAR); um grupo tratado com terapia celular e após noventa dias os dois grupos reavaliados com TCAR antes da eutanásia, totalizando para os dois grupos, cento e oitenta dias de doença instalada. As análises tomográficas mostraram que o tempo para que a doença intersticial seja estabelecida ocorre até três meses após a infusão de bleomicina. As provas histológicas corroboram a viabilidade do modelo testado e as análises imunohistoquímicas sugerem a migração das células mononucleares de medula óssea para os pulmões, bem como a presença de populações celulares que indicam provável reestruturação do parênquima pulmonar. / Fibrotic Interstitial pulmonary illnesses affect men, women and children, and presents bad prognostic, 3-5 years depending on the diagnostic confirmation, mainly in idiopathic pulmonary fibrosis. Recent studies demonstrate the capacity of the stem cells in differentiating into different cellular lineages and different tissues, as in heart, liver, gastrointestinal, nervous system and lung. The objectives of this study were to investigate the possibility to consider the swine as experimental model of fibrotic pulmonary disease and experimental stem cell therapy. Bleomycin sulphate was injected into the trachea to induce the pulmonary disease in control and treatment groups. High resolution computed scan (TCAR) was carried out in both groups after the confirmation of the disease. The tomographic analyses showed that the interstitial illness was established after three months of the bleomicine infusion. Histologic investigation revealed the viability of the tested model and the imunohistochemical analyses suggest the migration of the mononuclear cells to the lungs, as well as the presence of new cellular populations that would indicate probable reorganization of the pulmonary parenchyma.

Introduction : Les pneumopathies interstitielles diffuses (PID), incluant les fibroses pulmonaires idiopathiques (FPI), sont des pathologies rares et sévères. Dans moins de 20% des cas, un défaut moléculaire d'un gène du complexe des télomérases ou du surfactant est identifié. L'objectif de ce travail a été de documenter, dans une cohorte de patients pédiatriques et adultes présentant une PID sans cause identifiée, des variations des gènes codant les protéines du surfactant (SP), en particulier SP-A1, SP-A2 et SP-D. Méthodes : Les gènes SFTPA1, SFTPA2 et SFTPD, ont été séquencés et les variations identifiées ont été étudiées par des analyses fonctionnelles et tissulaires. Résultats : Dans une cohorte de 345 patients, une mutation hétérozygote du gène SFTPA1 (p.Trp211Arg) a été identifiée dans une large famille. Les patients présentaient des PID de l'enfant et des PID/FPI l'adulte, associées pour certains à des adénocarcinomes pulmonaires. La mutation W211R concernait un acide aminé invariant au cours de l'évolution et altérait la sécrétion de SP-A1 et l'expression tissulaire de SP-A. Cinq nouvelles variations de SFTPA2 ont aussi été identifiées. Au total, une variation de l'un des gènes du complexe du surfactant a été retrouvée pour 7,5% des patients. Conclusion : Cette étude a permis d'impliquer pour la première fois le gène SFTPA1 dans les PID de l'enfant et de l'adulte. L'association entre les mutations de SFTPA1 et SFTPA2 et un risque plus élevé de cancers pulmonaire soulève la question d'un conseil génétique complexe, et ouvre des hypothèses physiopathologiques nouvelles ciblant, entre autres, les anomalies de prolifération et de réparation de la surface alvéolaire. / Background: Idiopathic interstitial pneumonias (IIP) comprise a heterogeneous group of rare lung parenchyma disorders with high morbidity and mortality. In adults, the most common form of IIP is idiopathic pulmonary fibrosis (IPF). A genetic cause is identified in up to 20% of cases, telomerase and surfactant genes mutations being the first etiologies. This study aims to investigate the implication of the genes encoding the surfactant protein (SP), in particular SP-A1, SP-A2, and SP-D, in pediatric and adult patients with unexplained IIP.Methods: Surfactant genes, in particular SFTPA1, SFTPA2 and SFTPD, were sequenced. New variations were studied with functional and tissues experimentations. Results: The study involved 345 patients. A heterozygous missense mutation (p.Trp211Arg) in SFTPA1 was identified in a large family. The IIP were isolated or associated with an adenocarcinoma of the lung in patients with IPF. The W211R mutation involves an amino-acid that is invariant throughout evolution, impairs SP-A1 secretion, and is associated with an abnormal tissue expression of SP-A. Five new SFTPA2 mutations were also identified, and all together, a surfactant gene variation was described for 7.5% of the patients.Conclusion: This study has involved for the first time SFTPA1 in pediatric and adult IIP. The association of SFTPA1 and SFTPA2 mutations with an increased risk of lung cancer raises the issue of a complex genetic counseling in the involved families. It also brings out new pathophysiologic hypothesis such as aberrant proliferation and epithelial repair of the alveolar surface.

Master of Science / Department of Clinical Sciences / Barret J. Bulmer / Degenerative valve disease is the most common cardiac affliction facing our canine population. To date, canine research has focused on characterizing the disease itself and the histopathological features. Because of the ability to routinely repair or replace diseased valves in human medicine, research focus in humans has been on perfecting these techniques rather than elucidating etiology. The recent interest in valvular interstitial cells has been primarily due to their capacity to degrade collagen with the knowledge that disorganized collagen is a hallmark characteristic of degenerative valve disease. In this project, an easily reproducible cell culture protocol for canine valvular interstitial cells was developed. These cells were phenotyped by utilization of RT-PCR and immunocytochemistry. The use of these cells in a research project looking at response to endothelin exposure with and without protection of vitamin E is demonstrated as an example of the unlimited possibilities for these cells to elucidate not only the etiology of the disease process but also the response to therapy.

Les zones humides (ZH) alluviales constituent des secteurs à forte valeur écologique etpatrimoniale. Pour comprendre et prédire l’évolution à long terme de ces zones, en particulierface aux changements globaux, il apparaît primordial de traduire les perturbationshydrologiques attendues en termes de perturbations biologiques, afin d'estimer lesconséquences potentielles de ces changements pour la biodiversité et le fonctionnement globalde ces écosystèmes. Cette thèse s’articule donc autour de deux objectifs: proposer un modèleconceptuel de fonctionnement écologique des ZH soumises à des assèchements saisonniers etestimer la vulnérabilité écologique de communautés faunistiques vivant dans ces ZH, enconsidérant conjointement les peuplements benthiques et interstitiels (principalementCrustacés).Nos résultats démontrent que la morphologie des ZH (eau relativement courante, eaustagnante, apports phréatique) et leurs modes d’assèchement (formation de mouilles isoléesou assec complet) sont essentiels dans la structuration des communautés microbiennes etd’invertébrés et dans la réponse saisonnière de la biodiversité et des processus écologiques.L’intensification probable des étiages et des assèchements augmentera l’hétérogénéité spatialeet temporelle des habitats et leur richesse spécifique mais réduira la biodiversité globale desplaines alluviales (disparition des espèces rares). Ces étiages plus sévères modifieront lesprocessus écologiques, diminuant le recyclage de la matière organique et la qualité des eauxsuperficielles. Les recherches futures devraient s’orientées vers des stratégies et des moyensefficaces pour garantir la durabilité écologique des ZH. / Alluvial wetlands offer many ecological goods and services. In order to evaluate thelong term impact of global changes, it appeared essential to translate the hydrological changesin term of biological perturbation, so as to estimate the consequences on biodiversity andecosystem functioning. This PhD thesis focuses on two points: to propose a conceptual modelof ecological functioning of the wetlands that are subjected to seasonal drying and to estimatethe ecological vulnerability of aquatic invertebrate communities of the wetlands facing theincreasing drought risk, by considering both the benthic and interstitial assemblages (with aparticular interest on Crustaceans).Our main results demonstrate that wetlands’ morphology (running water, groundwaterupwelling, stagnant water) and their drying modalities (formation of isolated pools orcomplete drying) influence both the microbial and invertebrate community structures andcontrol the seasonal responses of biodiversity and ecological processes. The potential increasein the low flow period and drying intensity may increase the spatial and temporalheterogeneity of habitats and their specific richness but may decrease the global biodiversityof floodplain (rarefaction of sensible species). Severe low flow periods may also disturb theirecological functioning, reducing the organic matter recycling and the surface waters quality.Future studies should develop efficient strategies to support the ecological durability ofwetlands.

The subject of this thesis is preparation of CoCrFeMnNiNx high entropy mixtures via the methods of mechanical alloying and spark plasma sintering (SPS). Three series of specimens were fabricated in this thesis: samples milled in argon (benchmark materials), samples milled in nitrogen atmosphere (to observe their ability of nitrogen absorption) and samples microalloyed with CrN, FeN nitrides (to observe their dissociation into the solid solution potential). The fabricated powders and SPS compacts were subsequently observed by electron microscopy and their phase content by X-Ray diffraction (XRD) and elemental composition by EDS analysis were carried out. A method of reduction melting in inert atmosphere was used to determine the exact oxygen and nitrogen content in powders, while the respective particle size distribution measured by laser diffraction method. The influence of nitrogen content on the hardness of the samples was studied via the microhardness measured. After completing the process of mechanical alloying under the Nitrogen atmosphere was the maximal concentration of nitrogen in the structure 0,208% after 24 hours of milling (dependency on time was linear), which means, the method of milling under the Nitrogen atmosphere was successful. XRD of milled samples showed the existence of the only FCC single solid solution phase, while samples milled under the Nitrogen atmosphere showed the trend of the growth of the lattice parameter with the increasing nitrogen content. There was observed the presence of the chromium nitrides precipitates on the grain boundaries of the FCC phase in microalloyed samples. All specimen were contaminated by a mixture of metallic oxides and manganeese sulphides, which were present in the default manganeese powder. The greatest value of microhardness showed the duplex sample. The increase in values of microhardness (344 HV 0,3) in comparison with the standard sample (262,9 HV 0,3) was recorded on the samples milled under the nitrogen atmosphere, which conforms the positive influence of the nitrogen content on strength characteristics of this alloy.

Oxygen strengthened biomedical beta titanium alloys Beta titanium alloy Ti-35Nb-6Ta-7Zr-0.7O (wt. %) exhibits a low Young's modulus and high strength thanks to interstitial strengthening effect of oxygen. These mechanical properties are promising for implant production. In this thesis, the aforementioned alloy is thoroughly studied in conditions prepared by various thermomechanical treatments, namely: material after casting, hot compression testing, die-forging, hot rolling and finally, cold-swaged and annealed condition. Mechanical properties and microstructure of prepared conditions are studied with special interest of enhancing the fatigue performance that is poor after casting due to porosity and large grains. The hot rolling was shown to improve the fatigue performance significantly, mainly through grain refinement. It is shown that by combining the effects of cold-swaging and recrystallization annealing, extensive refinement of grain structure can be achieved. Industrial aspects of implant production by the studied methods were discussed. In the second part of the thesis, the possibility of lowering the Young's modulus by reducing the beta phase stability (by reducing of Nb and Ta content), while retaining high strength caused by high interstitial oxyen content is explored. Several alloys were...

Harnblase. Weiterhin bestärkt die beobachtete Kopplung die Hypothese, dass das Myofibroblastennetzwerk afferente Signale Verstärken und modulieren kann. Die Verbesserung des Verständnisses der Harnblasenfunktion ist von grundlegendem Interesse für die Behandlung von Miktionsstörungen. Eine Störung von mechanischer Stimulierbarkeit und Kopplung der suburothelialen Myofibroblasten kann eine Störung der Sensorik der Harnblase plausibel erklären und stellt einen neuen Therapieansatz dar.:1 Abkürzungsverzeichnis 2 Einführung 2.1 Hintergrund 2.2 Anatomische und physiologische Grundlagen der Harnblasenfunktion 2.2.1 Aufbau der Harnblasenwand 2.2.2 Terminologie und Klassifikation interstitieller Zellen der Harnblase 2.2.3 Phänotypisierung der suburothelialen Myofibroblasten 2.2.4 Funktionsweise der Myofibroblasten 2.2.5 Innervation des unteren Harntraktes und neuronale Kontrolle der Harnblasenfunktion 2.2.6 Lokale sensorische Netzwerke der Harnblase 3 Aufgabenstellung 4 Material und Methoden 4.1 Gewinnung und Zellkultur suburothelialer Myofibroblasten 4.2 Lösungen und Chemikalien 4.3 Calcium imaging 4.4 Datenanalyse 4.4.1 Erzeugung der FI-Ratio Datensätze 4.4.2 Automatische Fluoreszenz-Signal-Analyse 4.5 Aufbau und Anordnung der Experimente 4.5.1 Spontanaktivität 4.5.2 Mechanische Stimulation mittels Glasmikropipette 4.5.3 Mechanische Stimulation durch Scherstress 4.5.4 Hypoosmolare Stimulation 5 Ergebnisse 5.1 Spontane Kalziumaktivität humaner suburothelialer Myofibroblasten 5.2 Mechanische Stimulierbarkeit durch Druck mittels Glasmikropipette 5.2.1 Intrazelluläre Ausbreitung mechanisch induzierter Kalziumsignale 5.2.2 Interzelluläre Ausbreitung mechanisch induzierter Kalziumsignale in kultivierten humanen suburothelialen Myofibroblasten 5.3 Mechanische Stimulierbarkeit durch Scherstress 5.4 Mechanische Stimulierbarkeit durch osmotischen Stress 6 Diskussion 6.1 Beobachtete Kalziumsignale suburothelialer Myofibroblasten 6.2 Bedeutung der mechanischen Stimulierbarkeit 6.3 Identifikation der stretch-activated channels 6.4 Bedeutung der interzellulären Kopplung 6.5 Konzept zur Rolle der suburothelialen Myofibroblasten 6.6 Methodischer und experimenteller Ansatz 6.6.1 Selbst entwickelte Fluoresence Analysis Software 6.6.2 Methoden der mechanischen Stimulierbarkeit 6.7 Pathophysiologische Aspekte 7 Zusammenfassung der Arbeit 8 Literaturverzeichnis 9 Anlagen 9.1 Selbstständigkeitserklärung 9.2 Lebenslauf 9.3 Publikationen 9.4 Danksagung