La sténose valvulaire aortique (SVA) est une pathologie associée au vieillissement et aux facteurs de risque cardiovasculaire. Afin d’étudier la SVA et d’explorer de nouvelles thérapies, plusieurs modèles animaux ont été récemment développés, mais la plupart de ces modèles ciblent les mécanismes de développement de la SVA reliés à l’hypercholestérolémie. Le syndrome de Werner (WS) est une maladie caractérisée par un vieillissement prématuré. Récemment, il a été découvert que des souris mutantes ayant une délétion du domaine hélicase du gène Werner, responsable du WS, démontraient un profile hémodynamique typique de la SVA. De ce fait, nous avons émis l’hypothèse que ces souris pourraient développer une SVA plus rapidement que des souris de type sauvage. Nous avons donc étudié les effets cette mutation chez des souris WrnΔhel/Δhel, en comparant le taux de progression d’une SVA entre des souris WrnΔhel/Δhel (WrnΔhel) et des souris de type sauvage comme groupe contrôle. À la suite d’une diète riche en sucre et en gras sur une période de 24 semaines, les souris WrnΔhel ont démontré une diminution plus prononcée de leur aire de valve aortique (mesures échocardiographiques) que les souris contrôles, supportée par les analyses histologiques concernant la fibrose des valves aortiques. Les souris n’ont toutefois développé aucun signe évident d’athérosclérose comme l’infiltration de lipides ou l’inflammation, bien que certaines caractéristiques liées à la dysfonction endothéliale semblent être augmentées chez les souris WrnΔhel. D’autres mesures échocardiographiques indiquant une SVA, comme une hypertrophie du ventricule gauche dans le groupe WrnΔhel, ont été obtenues. Nous avons aussi observé des indices de vieillissement plus marqués quant aux analyses sanguines et de la moelle osseuse des souris WrnΔhel en comparaison avec les souris contrôles. Par conséquent, ce modèle expérimental de vieillissement pourrait être utilisé pour les études futures sur la SVA sans les principaux effets athérogéniques des autres modèles expérimentaux. / Aortic valve stenosis (AVS) is associated with aging and classical cardiovascular risk factors. Different animal models were recently developed to study AVS and explore new therapies, however, most of these models rely almost exclusively on hypercholesterolemia-related mechanisms for AVS development. Werner syndrome (WS) is a disorder characterized by premature aging. It was recently demonstrated that mutant mice with a deletion of the helicase domain of the Werner gene, the gene responsible for WS, showed hemodynamic profile typical of AVS. We therefore hypothesized that mice with the WrnΔhel deletion could develop AVS earlier than wild-type (WT) mice. We studied the effect of the WrnΔhel mutation by comparing the rate of progression of AVS in homozygous mutant versus WT mice. By twenty-four weeks on a high-fat/high-carbohydrate diet, WrnΔhel/Δhel (WrnΔhel) mice showed a stronger decrease of the aortic valve area measured by serial echocardiography than WT mice, supported by histological analyses of valve fibrosis but without developing major signs of atherosclerosis such as lipid infiltration or increased inflammation. Some features linked to endothelial dysfunction also appeared to be increased in WrnΔhel mice. Other echocardiographic measurements were typical of AVS, such as left ventricle hypertrophy in the WrnΔhel group. We also observed stronger aging properties from WrnΔhel mice bone marrow and blood analyses compared to the WT group. Consequently, this experimental aging model could be used for AVS research without the major confounding atherogenic effects of other experimental models.

The objectives of this thesis are: a) To examine racial/ethnic differences in coronary artery calcification (CAC) and CAD, between symptomatic South Asians and Caucasians, matched for age, gender and conventional cardiovascular risk factors, b) To assess, using a meta-analysis model, the natural history of and stability of measurements of coronary artery calcium scoring (CACs) based on data collected from two large published trials: St Francis and EBEAT, c) To investigate the prevalence of coronary artery calcification in individuals with CT evidence for AVC, mitral valve calcification (MAC) or of both of them (AVC+MAC), d) To assess any potential association between premature CAD (<55 years in first-degree male relatives and <65 years in first-degree female relatives) and CAC in a large cohort of asymptomatic individuals. We found that coronary artery calcification is more extensive and diffuse in symptomatic patients of South Asian ethnic origin as compared to Caucasians, despite similar conventional risk factors for CAD. This is more evident in those >50 years of age, suggesting potential genetic or other risk factors yet to be determined. The natural history of coronary artery calcification was overtime progression in the majority of subjects, irrespective of gender. The higher variability in RCA measurements could be related to the low baseline CACs or exaggerated movement of the right side atrioventricular ring, whereas those for LCA brances are influenced by the branch allocation of the CACs. Valve calcification is not isolated but involve also and the coronary arteries. The presence of calcification in the aortic valve or combined aortic and mitral valves predicted coronary artery calcification. Additionally patients in whom both valves have become calcified tend to have severe coronary artery calcification. And finally, there is no relationship between the prevalence and extent of coronary artery calcification and the presence of family history of coronary heart disease in asymptomatic individuals with none of the conventional risk factors for atherosclerosis.

Aortic stenosis is the most common acquired heart valve condition. Open-heart surgical aortic valve replacement is an effective treatment for patients who receive it. However, approximately one-third of patients who require this treatment do not receive it due to the risks associated with the surgery. Percutaneous aortic valve replacement (PAVR) is a minimally invasive technique that can replace the aortic valve of patients contraindicated for open-heart surgery. Although PAVR is now entering clinical practice, a closed bore hybrid x-ray/MRI (CBXMR) imaging system is under development to improve the safety and efficacy of PAVR. This system will harness the complementary strengths of x-ray imaging (surgical tool/vascular imaging) and MRI (cardiac soft tissue contrast) to deploy a bioprosthesis in the aortic annulus. An x-ray C-arm will be placed about 1 m from the entrance of the MRI scanner to facilitate smooth intermodality patient transfer during the procedure. The performance of a rotating-anode x-ray tube in the magnetic fringe field of a 1.5 T MRI scanner was investigated. A rotating-anode x-ray tube provides the fluoroscopy and angiography needed for PAVR. The magnetic fringe field can affect the ability of the x-ray tube to dissipate heat. It was shown that the fringe field perpendicular to the anode rotation axis can reduce anode rotation frequency. These effects can limit the maximum permissible power that can be safely dissipated on the anode track during a single exposure. In the fringe field strengths at the C-arm position (4-5 mT), anode rotation frequency only decreased by about 1%, which will have negligible impact on tube heat loadability. The fringe field can cause a field of view shift. The field of view shifted by approximately 3 mm, which can be corrected by active magnetic shielding and further collimation. An active magnetic shielding system was constructed that can correct focal spot deflection. These results are facilitating the construction of a prototype CBXMR system, the goal of which is to improve success rates for PAVR procedures.

[pt] A substituição de válvula aórtica por cateter (Transcatheter Aortic Valve Replacement, TAVR) tornou-se uma poderosa alternativa para pacientes com estenose aórtica e com alto risco de serem submetidos à cirurgia tradicional de peito aberto. O conhecimento da distribuição da pressão, bem como a tensão cisalhante na superfície da aorta podem ajudar a identificar regiões críticas, onde o processo de remodelamento da aorta pode ocorrer. O objetivo do presente trabalho é avaliar numericamente a influência do posicionamento do orifício da válvula protética no campo de escoamento. O estudo foi realizado com base em um paciente submetido a TAVR. Um modelo 3D foi gerado a partir de angiotomografia e de segmentação de imagens da aorta. Dados experimentais obtidos anteriormente na mesma geometria indicaram que o fluxo do jato através da válvula de entrada é de natureza turbulenta. O escoamento foi determinado numericamente com o software comercial FLUENT. A turbulência foi modelada com o modelo de dois equações k-omega SST. Para representar um fluxo pulsátil, foram impostos diferentes fluxo de massa na entrada da válvula. Para todas as vazões investigadas, obteve-se um padrão de escoamento semelhante. Mostrou-se que uma pequena variação dos ângulos de inclinação pode modificar a natureza do fluxo, deslocando a posição dos vórtices e alterando a localização das regiões de alta tensão de cisalhamento, assim como de alta pressão, na superfície interna da aorta. Mostrou-se também que um aumento da intensidade da turbulência na entrada diminui os valores de tensão cisalhante e de pressão nas paredes da aorta. Essas características hemodinâmicas podem ser relevantes no processo de remodelação aórtica e os estresses mecânicos podem influenciar na durabilidade da prótese valvular. / [en] Transcatheter Aortic Valve Replacement (TAVR) has become a powerful alternative for patients with aortic stenosis and a high surgical risk to face a traditional open chest surgery. The knowledge of the pressure distribution as well as shear stress at the aortic surface may help identify critical regions, where aortic remodeling process may occur. The purpose of the present work is to evaluate numerically the influence of the positioning of the prosthetic valve orifice in the flow field. The study was carried out on the basis of a particular patient who had undergone a TAVR. A 3D model was generated from computed tomography angiography and image segmentation of the aorta. Experimental data previously obtained in the same geometry indicated that the jet flow through the inlet valve is turbulent flow. The flow field was numerically determined with the commercial software Fluent. The turbulence was modeled with the two-equation k-omega SST model. To represent a pulsatile flow, different mass flow rates were imposed at the inlet valve. Similar flow pattern was observed for all flow rates investigated. It was shown that small variations of the tilt angle can modify the nature of the flow, displacing the position of the vortices and altering the location of high shear stress, as well as high pressure, at the aortic inner wall. It was also shown that an increase of the turbulent intensity at the entrance decreases the values of shear stress and pressure on the walls. These hemodynamic features may be relevant in the aortic remodeling process and the mechanical stresses may influence the durability of the valve prosthesis.

Cette thèse traite plusieurs aspects scientifiques inhérents à la simulation numérique de problèmes d'interaction fluide-structure impliquant de fines membranes déformables. Deux cas spécifiques relatifs à la biomécanique cardiovasculaire sont considérés : l'interaction de l'écoulement sanguin avec la valve aortique (qui se produit à l'échelle macroscopique), et l'interaction de la membrane des globules rouges avec ses fluides interne et externe (qui se produit à l'échelle microscopique). Dans les deux cas, le couplage fluide-structure est géré par l'intermédiaire d'un formalisme de frontières immergées, en représentant la membrane par un maillage Lagrangien se mouvant au travers d'un maillage fluide Eulérien. Lorsque l'on traite la dynamique des globules rouges, la membrane est considérée comme étant une structure sans masse et infiniment fine. La première question à laquelle on s'intéresse dans cette thèse est la manière de modéliser la microstructure complexe de la membrane des globules rouges. Un moyen possible pour caractériser un modèle de membrane adapté est de simuler l'expérience des pinces optiques, qui consiste en une configuration expérimentale bien contrôlée qui permet d'étudier la mécanique individuelle d'un globule rouge isolé dans une large gamme de déformations. Plusieurs modèles pertinents sont identifiés, mais les caractéristiques de déformation mesurées durant l'expérience des pinces optiques se révèlent n'être pas assez sélectives pour être utilisées dans un contexte de validation. Des mesures de déformation additionnelles sont proposées, qui pourraient permettre une meilleure caractérisation de la mécanique de la membrane des globules rouges. En ce qui concerne les configurations macroscopiques, une méthode numérique innovante est proposée afin de gérer des simulations numériques de membranes 3D continues, en conservant le formalisme de frontières immergées. Dans cette méthode, appelée méthode des frontières immergées épaisses, la membrane a une épaisseur finie. La précision et la robustesse de la méthode sont démontrées par l'intermédiaire d'une variété de cas tests bien choisis. La méthode proposée est ensuite appliquée à un problème d'interaction fluide-structure réaliste, à savoir l'interaction d'un écoulement (sanguin) pulsé avec une valve aortique biomimétique. Une étude combinée expérimentale et numérique est menée, montrant que la méthode est capable de capturer la dynamique globale de la valve, ainsi que les principales caractéristiques de l'écoulement en aval de la valve. Tous les développements ont été effectués dans le solveur YALES2BIO (http://www.math.univ-montp2.fr/~yales2bio/) développé à l'IMAG, qui est donc disponible pour toutes autres améliorations, validations et études applicatives. / This thesis deals with several scientific aspects inherent to the numerical simulation of fluid-structure interaction problems involving thin deformable membranes. Two specific cases relevant to cardiovascular biomechanics are considered: the interaction of the blood flow with the aortic valve (which occurs at the macroscopic scale), and the interaction of the red blood cells membrane with its inner and outer fluids (which occurs at the microscopic scale). In both cases, the fluid-structure interaction coupling is handled using an immersed boundary formalism, representing the membrane by a Lagrangian mesh moving through an Eulerian fluid mesh.When dealing with red blood cells dynamics, the membrane is considered to be an infinitely thin and massless structure. The first question which is addressed in the present thesis work is how to model the complex microstructure of the red blood cells membrane. A possible way to characterize a suitable membrane model is to simulate the optical tweezers experiment, which is a well-controlled experimental configuration enabling to study the individual mechanics of an isolated red blood cell in a large range of deformation. Some relevant membrane models are identified, but the deformation characteristics measured during the optical tweezers experiment reveal to be not selective enough to be used in a validation context. Additional deformation measurements are proposed, which could allow a better characterization of the red blood cell membrane mechanics.Regarding the macroscopic configurations, an innovative numerical method is proposed to handle numerical simulations of 3D continuum membranes, still within the immersed boundary formalism. In this method, called immersed thick boundary method, the membrane has a finite thickness. The accuracy and robustness of the method are demonstrated through a variety of well-chosen test cases. Then, the proposed method is applied to a realistic fluid-structure interaction problem, namely the interaction of a pulsatile (blood) flow with a biomimetic aortic valve. A combined experimental and numerical study is led, showing that the method is able to capture the global dynamics of the valve, as well as the main features of the flow downstream of the valve.All the developments were performed within the YALES2BIO solver (http://www.math.univ-montp2.fr/~yales2bio/) developed at IMAG, which is thus available for further improvements, validations and applicative studies.

Made available in DSpace on 2015-07-22T20:50:12Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-25. Added 1 bitstream(s) on 2015-08-11T03:25:30Z : No. of bitstreams: 1 Publico-00375a.pdf: 1753829 bytes, checksum: b27d39899f37ed74cdddcf1f909cb0f6 (MD5). Added 1 bitstream(s) on 2015-08-11T03:25:30Z : No. of bitstreams: 2 Publico-00375a.pdf: 1753829 bytes, checksum: b27d39899f37ed74cdddcf1f909cb0f6 (MD5) Publico-00375b.pdf: 1511839 bytes, checksum: 43d714b64137888aafbeea17f2c9bec1 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / OBJETIVO: A troca valvar aórtica é procedimento rotineiro, envolve substituição da valva nativa/prótese. Na maioria destes pacientes o risco é aceitável, porém, em alguns casos, o risco predito pode justificar contra-indicação. O implante de valva aórtica minimamente invasivo transcateter tem se mostrado viável e com menor morbi-mortalidade. O objetivo deste trabalho foi desenvolver bioprótese aórtica nacional, montada em cateter, para implante em posição aórtica. MÉTODO: Após desenvolvimento em animais, 14 pacientes com EuroSCORE elevado foram submetidos ao implante. A idade média dos pacientes foi 75,5 anos. O Euroscore logístico médio foi 43,7 %. Quatro pacientes apresentavam disfunção de biopróteses e o restante, estenose aórtica grave com gradiente de pico médio de 79,7 mmHg. Após minitoracotomia e sob controle ecocardiográfico e fluoroscópico, cateter-balão foi posicionado sobre posição aórtica e insuflado. Após, segundo cateter-balão, com endoprótese valvada, foi posicionado e liberado sob alta frequência ventricular ou hipotensão controlada por drenagem sanguínea. Controles angiográficos e ecocardiográficos foram realizados e pacientes encaminhados para UTI. RESULTADOS: O implante foi possível em 13 casos. Não ocorreram óbitos intraoperatórios. A mortalidade geral foi 42%. O gradiente valvar aórtico reduziu de forma significativa para 25,0 mmHg em média, a função ventricular esquerda apresentou melhora significativa nos primeiros 7 dias de evolução. A regurgitação aórtica perivalvar esteve presente na maioria dos casos, porém foi predominantemente leve. A contagem plaquetária reduziu de forma significativa após o procedimento, recuperando seu valor inicial apenas após 30 dias. CONCLUSÃO: O implante de bioprótese transapical montada em cateter mostrou ser procedimento factível. Detalhes técnicos e a curva de aprendizado demandam discussão. / OBJECTIVE: The aortic valve replacement is a routine procedure, and involves replacement of the native valve/prosthesis. In most of the patients who undergo such procedure the risk is acceptable, but in some cases, such risk can justify contraindication. The minimally invasive transcatheter aortic valve implantation has been viable, with lower morbidity and mortality. The aim of this study was to develop a national catheter-mounted aortic bioprosthesis for aortic position implant. METHOD: After animal studies, 14 patients with high EuroSCORE underwent transcatheter aortic valve implantation. Median age was 75,5 years. Median Logistic EuroSCORE was 43,7%. Four patients presented with disfunctional bioprosthesis, remainig ones presented calcified aortic stenosis (peak transvalvular gradient 79,7 mmHg). After minithoracotomy and under echocardiograph and fluoroscopic control, a balloon catheter was placed on aortic position and inflated. After, a second balloon with valved endoprosthesis was positioned and released under high ventricular rate. Echocardiograph and angiograph controls were performed and the patients were referred to ICU. RESULTS: Implant was feasible in 13 cases. There were no intra-operative deaths. Mortality was 42%. Median peak transvalvular aortic gradient reduced to 25,0 mmHg, and left ventricular function improved in the first 7 post operative days. Paravalvular aortic regurgitation was mild and present in most cases. Platelet count significantly decreased after procedure, but returned to normal values after 30 days. CONCLUSION: The transapical implantation of catheter-mounted bioprosthesis was a feasible procedure. Technical details and learning curve require further discussion. / TEDE / BV UNIFESP: Teses e dissertações

A necessidade de cirurgia para substituição da aorta ascendente e valva aórtica com reimplante coronariano em grupos especiais de pacientes onde a anticoagulação é indesejável, como nos idosos, é crescente nos últimos anos. Este estudo objetiva desenvolver e avaliar em modelo animal o comportamento de um conduto valvado aórtico feito com pericárdio bovino tratado pelo glutaraldeído (CVAP). Para tanto, CVAPs foram implantados em 8 ovinos jovens e explantados após serem sacrificados com 150 dias de pós-operatório. Realizou-se estudo angiográfico e hemodinâmico no préoperatório e antes do explante. Ecodopplercardiogramas foram realizados nos dias 30 e 150 de pós-operatório (teste) e também em 5 ovinos não operados (controle). Após explantados, submetemos os CVAPs à avaliação macroscópica, radiológica e histológica por microscopia óptica e eletrônica de transmissão. A análise estatística foi feita com teste não-paramétrico de Mann-Whitney, teste não-paramétrico de Wilcoxon e teste exato de Fisher. O nível de significância utilizado foi 5%. Na análise hemodinâmica houve acréscimo (p>0,05) das pressões arterial e capilar pulmonar entre os dias 0 e 150. Na análise ecodopplercardiográfica, o grupo teste apresentou valores maiores (p>0,05) dos diâmetros diastólicos e sistólicos do ventrículo esquerdo. No grupo teste entre os dias 30 e 150 houve acréscimo (p>0,05) de: peso, espessura das paredes do ventrículo esquerdo, gradiente transvalvar máximo, gradiente transvalvar médio, diâmetro diastólico do ventrículo esquerdo e decréscimo da fração de ejeção. Dois animais com endocardite podem explicar essas diferenças. A macroscopia demonstrou calcificação de grau variável, além de abaulamento na região dos seios em todos os CVAPs, sem aumento de diâmetro. A microscopia óptica revelou dados similares ao da literatura com o uso do pericárdio bovino tratado pelo glutaraldeído. A quantificação realizada com a microscopia eletrônica de transmissão mostrou percentual maior (p>0,05) de colágeno nos seios e nas cúspides e maior conteúdo hídrico na porção mais distal do CVAP. Estes dados indicam que a bioprótese aqui estudada permite a realização desse tipo de experimento no modelo proposto e que os resultados hemodinâmicos encontrados se assemelham aos parâmetros fisiológicos. / The need for replacement surgery of the ascending aorta and aortic valve with coronary reimplantation in special patient groups where anti-coagulation in undesirable, such as elderly, is increasing in the past years. This study aims to develop and evaluate the performance of an aortic valved conduit made with glutaraldehyde treated bovine pericardium (AVCP) in an animal model. Therefore, AVCPs were implanted in 8 young ovine and explanted after being euthanized at 150 days of the post-operative period. An angiographic and hemodynamic study was performed at pre-operative and prior the explant. EchoDoppercardiograms were performed at day 30 and 150 of post-operative (test) as well as in 5 non-operated on ovines (control). Following the explant, AVCPs were submitted to a macroscopical, radiological and histological evaluation by optic and electronic transmission microscopy. A statistics analysis was performed with a Mann-Whitneys nonparametric test, Wilcoxons non-parametric test and Fishers exact test. The significance level used was 5%. In the hemodynamic analysis an increase (p>0,05) of arterial and pulmonary capillary pressure occurred between day 0 and 150. In the echoDoppercardiographic analysis, the test group presented higher values (p>0,05) in the diastolic and systolic diameters of the left ventricle. In the test group, between day 30 and 150, occurred an increase (p>0,05) of weight, thickness of the left ventricle walls, maximum transvalvar gradient, medium transvalvar gradient, left ventricle diastolic diameter and a decrease in the ejection function. Two animals with endocarditis could explain those differences. Macroscopy showed a calcification in variable degrees besides a bulging of the sinus region in all AVCPs, without an increase of diameter. Optic microscopy revealed data similar to literature with the use of glutaraldehyde treated bovine pericadium. The quantification performed by electronic transmission microscopy showed a higher percentage (p>0,05) of collagen in sinus and cusps, and a higher watery content in the most distal portion of the AVCP. These data indicate that the bioprosthesis hereby studied allows the performance of this kind of experiment in the proposed model and that the hemodynamic outcomes found are similar to physiological parameters.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2016-08-12T11:35:34Z No. of bitstreams: 1 rafaelladesouzasalomaozanette.pdf: 856423 bytes, checksum: c95ae99e7ef9f490e08fd22c64f7237f (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-08-15T13:04:26Z (GMT) No. of bitstreams: 1 rafaelladesouzasalomaozanette.pdf: 856423 bytes, checksum: c95ae99e7ef9f490e08fd22c64f7237f (MD5) / Made available in DSpace on 2016-08-15T13:04:26Z (GMT). No. of bitstreams: 1 rafaelladesouzasalomaozanette.pdf: 856423 bytes, checksum: c95ae99e7ef9f490e08fd22c64f7237f (MD5) Previous issue date: 2016-02-25 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O coração é um órgão vital, que bombeia o sangue permitindo a sua circulação pelo corpo. A valva aórtica, e as suas estruturas de apoio ventriculares, formam a peça central do coração. Todas as câmaras do coração estão relacionadas diretamente à valva e seus folhetos são incorporados diretamente no esqueleto cardíaco. Falhas dos folhetos aórticos são as mais comuns entre as doenças relacionadas às valvas do coração, tendo como consequência o impacto negativo na vida do paciente, bem como nas despesas dos sistemas de saúde em todo o mundo. O desenvolvimento de materiais capazes de substituir com eficácia o tecido danificado da valva aórtica é de grande interesse na medicina regenerativa e engenharia de tecidos. As valvas mecânicas e biológicas têm sido amplamente estudadas, mas várias questões relacionadas com a integração ao hospedeiro ainda não foram resolvidas. Promissores protocolos para descelularização de tecidos de valva cardíaca têm sido investigados como uma alternativa para a preparação do material de substituição e para ser empregado como um substrato para a recelularização da matriz. No entanto, alguns protocolos de descelularização utilizados hoje em dia têm algumas limitações, uma vez que ainda não existe um método que permita a descelularização e ao mesmo tempo mantenha a estrutura da matriz extracelular para a posterior recelularização, evitando a rejeição após a implantação e futura substituição da prótese. Assim, o presente trabalho visa a obtenção de uma matriz biológica que satisfaça esses requisitos, por meio da descelularização dos folhetos aórticos de suínos e sua recelularização com células-tronco da polpa dentária humana. Foram testados três protocolos de descelularização, sendo que os protocolos que utilizaram tripsina foram reprodutíveis e forneceram matrizes com menor quantidade de DNA, quando comparados com o protocolo que utilizou apenas detergente e não foi reprodutível. As células-tronco podem ser isoladas de dentes decíduos (SHEDs) humanos e pesquisas apontam sua importância na medicina regenerativa visando à reconstrução de folhetos aórticos. Nesse trabalho, as SHEDs foram obtidas e caracterizadas fenotipicamente, sendo positivas para marcadores mesenquimais e embrionários e negativas para marcadores hematopoiéticos, além de apresentarem, in vitro, potencial de diferenciação osteogênica. Tais células foram então cultivadas em placas de petri com as matrizes descelularizadas, porém não houve sucesso na recelularização. Para tentar contornar esse problema, foi construído um biorreator a fim de aumentar a eficiência da recelularização do tecido, no entanto os resultados estão sendo testados. Vários pontos ainda devem ser abordados a fim de superar os obstáculos da técnica de descelularização do tecido para obter sucesso na recelularização. / Heart is a vital organ that pumps blood allowing its circulation through the body. The aortic valve and their ventricular support structures form the centerpiece of the heart. All chambers of the heart are directly related to the valve, and its leaflets are directly incorporated into the heart skeleton. Amongst heart valves diseases, failures in the aortic leaflets are the most common ones, leading to negative impacts on patients’ life, as well as increases on the costs for health systems worldwide. For regenerative medicine and tissue engineering, the development of materials capable of effectively replace damaged aortic valve tissue is of great interest. The mechanical and biological valves have been widely studied, but several issues related to integration to the host have not yet been resolved. Promising decellularization protocols for tissue heart valve have been investigated as an alternative for the preparation of replacement material and to serve as a substrate for matrix repopulation. However, some of the decellularization protocols used today have some limitations, since until now there is still no method which can achieve a complete decellularization while maintaining the structure of extracellular matrix for later repopulation, avoiding rejection after implantation and future replacement of the prosthesis. Thus, the present work aims at obtaining a biological matrix that satisfies such requirements through the decellularization of pigs aortic leaflets and its repopulation with stem cells isolated from human dental pulp. So far, we tested three decellularization protocols. The protocols using trypsin were shown reproducible and yielded a smaller amount of DNA, when compared to the protocol where only detergent was employed, which was also not reproducible. About the stem cells, they can be isolated from human deciduous teeth (SHEDs), with studies indicating its importance in regenerative medicine aimed at the reconstruction of aortic leaflets. In this work, SHEDs were obtained and phenotypically characterized, being positive for mesenchymal and embryonic markers and negative for hematopoietic markers, in addition to presenting in vitro osteogenic differentiation potential. The SHEDs were then cultured in Petri plates with the decellularized matrices, however there was no success in the matrix repopulation. In an attempt to overcome such problem, a bioreactor was built in order to increase the tissue repopulation efficiency, however the results are being tested. Several points still need to be addressed in order to overcome the obstacles of tissue decellularization technique and then achieve a successful recellularization.

Orientador: Maria Cecilia Bueno Jayme Gallani / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-04T09:44:49Z (GMT). No. of bitstreams: 1 Padilha_KatiaMelissa_M.pdf: 6172357 bytes, checksum: b3712bccfb1c3bfe26b9df7e8aee7cf6 (MD5) Previous issue date: 2005 / Resumo: Este trabalho teve como objetivo verificar as propriedades psicométricas do IDCV- Instrumento para mensuração do Impacto da Doença no Cotidiano do Valvopata no que se refere a: - validade de conteúdo; validade de constructo, por meio da análise fatorial e da validade divergente correlacionando-o com o IQV - Índice de Qualidade de Vida de Ferrans e Powers; -validade de critério correlacionando-o com a Medida Geral de Impacto da Doença e com o IEQ - Questionário para Avaliação do Impacto de Doenças Crônicas; e - consistência interna. A população deste estudo foi constituída por 120 pacientes valvopatas, atendidos no Ambulatório de Cardiologia do HCIUNICAMP. Os resultados obtidos foram submetidos às análises: descritiva; análise fatorial, correlação de Pearson e alfa de Cronbach. A análise fatorial gerou quatro fatores que explicaram em 58% a variância nas respostas: 1. Impacto físico da doença (sintomas); 2.Impacto da doença nas atividades cotidianas; 3.Impacto social e emocional da doença e 4. Adaptação à doença. Houve correlação que variou de fraca a moderada magnitude entre o escore total do IDCV e de seus fatores (3 e 4) e o IQV, comprovando sua validade divergente. Verificou-se correlação de forte magnitude entre o escore total do IDCV e o IEQ, de fraca a moderada magnitude entre os fatores 1, 2 e 3 do IDCV e o IEQ e entre o escore total do IDCV e de seus fatores 2 e 3 e a Medida Geral de Impacto da Doença, comprovando sua validade de critério. Foi constatado coeficiente alfa de Cronbach = 0,74, o que evidência a consistência interna do instrumento. Os resultados do presente estudo confirmam a validade de constructo e de critério, e apontam para consistência interna do IDCV. Estudos futuros são necessários para confirmar sua confiabilidade e fornecer maiores subsídios para a melhor compreensão do significado das dimensões do impacto mensuradas pelo IDCV, bem como para avaliar a responsividade do instrumento a intervenções de saúde. Apesar das características do estudo que impedem a generalização dos resultados e ampla recomendação do uso do IDCV, pode-se afirmar que este instrumento representa um passo inicial importante nos esforços para a maior compreensão sobre a vivência com a valvopatia e a eficácia das intervenções de saúde implementadas neste contexto / Abstract: The aim of this work was to verify the psychometric properties of the IDCV -Instrument for Measuring the Impact of Illness in the Daily Life of the valve heart in terms of: -content validity construct validity, by means ofthe factorial analysis and analysis of the divergent validity correlating it with the Life Quality Index (IQV) and - criterion validity, correlating it with the General Measure of Impact of Illness and with the Illness Effects Questionnaire (IEQ) - reliability.The subjectsthis studywere 120 patients with valve heart disease, who were attended at in the Cardiology Clinic at HC/UNICAMP. Data were submitted to the following analyses: descriptive, factorial analysis, Pearson's correlation coefficient and Cronbach' s Alpha coefficient. The factorial analysis generated four factors that explained in 58% the answers variance: 1. Physical impact of the illness (symptoms); 2.Impact of illness in the daily activities; 3.Social and emotional impact of illness and 4. Adaptation to illness. Correlation ranged from weak to moderate magnitude between the IDCV total score, and its factors (3 and 4), and between IDCV and the IQV, pointing to divergent validity. Strong magnitude correlation was verified between the IDCV total score and the IEQ; weak to moderate magnitude was verified between factors 1, 2 and 3 of the IDCV and the IEQ, and among the IDCV total score (its factors 2 and 3) and General Measure of Impact of Illness to, pointing to criterion validity. It was observed a Cronbach's Alpha of 0,74, which points to the internal consistency of instrument. The results of the current study confirm the construct and criterion validity, and points to internal consistency of the IDCV. Future studies are necessary to confirm its reliability and to provide more informations for a better understanding of the meaning of the dimensions in the impact measured by the IDCV, as well as to evaluate the response ofthe instrument to health interventions. Although this study presents characteristics that do not allow the generalization of the results and a strong recommendation for using the IDCV, it can be affirmed that this instrument represents an important first step in the efforts for an understanding of the experience with the vale heart disease and the effectiveness of the health interventions implemented in this context / Mestrado / Enfermagem e Trabalho / Mestre em Enfermagem

Introduction: La surcharge de pression ventriculaire augmente à l’exercice chez les patients avec une sténose de valve aortique (SVA). Lorsqu’il n’y a aucun symptôme apparent, il est cependant difficile d’indiquer l’intervention chirurgicale en utilisant seulement les indices de surcharge de pression ventriculaire. D’autres paramètres, tels que la dispersion de la repolarisation ventriculaire (d-QT), qui augmentent avec le gradient de pression transvalvulaire (GPT), n’ont pas été étudiés dans la SVA. L’objectif de l’étude était de déterminer le modèle de réponse du segment QT et de la d-QT à l’épreuve d’effort chez des enfants avec une SVA congénitale modérée afin d’évaluer l’impact de la surcharge de pression ventriculaire selon une perspective électrophysiologique. Matériel et méthodes: 15 patients SVA modérés ont été comparés à 15 sujets contrôles appariés pour l’âge (14.8±2.5 ans vs. 14.2±1.5 ans) et pour le sexe (66,7% de sujets mâles). Tous les sujets ont fait une épreuve d’effort avec enregistrement électrocardiographique à 12 dérivations. Le segment QT a été mesuré à partir du début du complexe QRS jusqu’à l’apex de l’onde T (QTa) au repos, à l’effort maximal ainsi qu’après 1 et 3 minutes de récupération. La longueur du segment QT a été corrigée selon l’équation de Fridericia et la d-QT a été calculée. Résultats: La longueur du segment QT corrigée (QTc) était similaire au repos entre les groupes d’étude, mais était significativement élevée chez les SVA en comparaison avec le groupe contrôle à l’effort maximal (p=0.004) ainsi qu’après 1 (p<0.001) et 3 (p<0.001) minutes de récupération. Une interaction significative a été identifiée entre les groupes pour la d-QT (p=0.034) et les tests post hoc ont révélé une différence significative seulement au repos (p=0.001). Conclusions: Les anomalies de repolarisation ventriculaire peuvent être révélées par l’évaluation de la repolarisation électrique lors de l’épreuve d’effort chez les SVA modérées asymptomatiques. L’utilisation de la réponse du QT à l’effort pourrait être bénéfique pour l’optimisation de la stratification du risque chez ces patients. / Introduction: Pressure overload increases in patients with moderate aortic valvular stenosis (AVS) during acute exercise. In the absence of symptoms, it remains difficult however, to discriminate patients for surgery based only on pressure overload. Other parameters, such as the dispersion of ventricular repolarisation (d-QT), which reportedly increases with the transvalvular pressure gradient, have not been fully studied in AVS. The aim of the study was to determine the pattern of QT and d-QT response to exercise testing in children with moderate AVS to evaluate the impact of pressure overload from an electrophysiological perspective. Materials and methods: 15 moderate AVS patients were compared to 15 controls paired for age (14.8±2.5 vs. 14.2±1.5 years old) and gender (66.7% male). All subjects underwent exercise stress testing with 12-lead ECG recording. QT was measured from the onset of QRS to the apex (QTa) at rest, peak exercise, 1 and 3 minutes of recovery. QT was corrected using the Fridericia equation and d-QT was calculated. Results: Resting QTc was similar among the study groups, but increased significantly in AVS in comparison to the control group at maximal effort (p=0.004) and after 1 (p<0.001) and 3 (p<0.001) minutes of recovery. A significant interaction was identified between groups for d-QT (p=0.034) and post-hoc tests revealed a significant difference only at rest (p=0.001). Conclusions: Ventricular repolarisation abnormalities can be unmasked and highlighted by the assessment of electrical repolarisation during exercise challenge in asymptomatic moderate AVS. Using QT response to exercise could be beneficial for better optimisation of risk stratification in these patients.