Crianças pequenas representam um grupo desafiador no transplante renal. O estudo analisa os resultados, do ponto de vista cirúrgico, do transplante renal em crianças com peso inferior a 15 kg utilizando o acesso cirúrgico extraperitoneal. Métodos: Foram revisados retrospectivamente os prontuários de 62 crianças com peso inferior a 15 kg submetidas a transplante renal entre 1998 e 2010, utilizando o acesso extraperitoneal e anastomose dos vasos renais dos doadores com a aorta ou artéria ilíaca comum e com a veia cava inferior ou ilíaca comum dos receptores. O ureter foi anastomosado à bexiga pela técnica extravesical de Lich- Grégoir. Resultados: Dos 62 transplantes, 32 enxertos (51,6%) eram provenientes de doadores vivos e 30 (48,4%) de doadores falecidos, sendo 28 deles pediátricos. A média de idade no transplante foi de 3,7 ± 2,2 anos (1 a 12), e o peso médio, de 12,3 ± 2,1 kg (5,6 a 14,9), sendo que 10 tinham peso inferior a 10 kg. Em 10 crianças (16,1%) o transplante foi preemptivo e em 5 (8,1%) havia trombose do sistema venoso prévio ao transplante. Em 1 e 5 anos, a sobrevida do paciente foi de 93,2% e 84,2% e a sobrevida do enxerto de 85,2% e 72,7%, respectivamente, sem diferença entre doadores vivos e falecidos. A função do enxerto com doador vivo foi melhor em 1 e 3 meses, mas a partir do 6o mês foi similar. Houve 6 complicações vasculares, sendo 4 tromboses vasculares, 1 laceração e 1 estenose de artéria renal e 2 coleções líquidas. Houve 17 perdas de enxerto, 6 por morte, sendo 5 com enxerto funcionante, 5 por complicações cirúrgicas, 3 por rejeição crônica e 3 por recorrência da doença de base. Conclusão: O acesso extraperitoneal é uma técnica válida no transplante renal de crianças com peso inferior a 15 kg, assegurando boa sobrevida do paciente e do enxerto e aceitável taxa de complicações, independentemente do tipo de doador, se vivo ou falecido, ou do tamanho do enxerto, se de adulto ou de criança. / Small children are a challenging group for kidney transplantation. This study analyzes the results of kidney transplantation in children weighing less than 15 kg using the extraperitoneal surgical access. Methods: A retrospective review of the records of 62 children weighting less than 15 kg was done. The kidney transplantation were performed between 1998 and 2010 using the extraperitoneal access and anastomosis of the renal vessels of donors to the aorta or common iliac artery and to the inferior vena cava or common iliac vein of the recipients. The ureter was anastomosed to the bladder using the Lich-Grégoir extravesical technique. Results: Thirty-two (51.6%) grafts of the 62 transplants were from living donors and 30 (48.4%) from deceased donors, 28 of them pediatric. The mean age at transplantion was 3.7 ± 2.2 years (1 to 12), and the mean weight, 12.3 ± 2.1 kg (5.6 to 14.9), and 10 of them weighed less than 10 kg. In 10 children (16.1%) the transplant was preemptive. Five 5 (8.1%) children presented previous thrombosis of the venous system. At 1 and 5 years, patient survival was 93.2% and 84.2% and graft survival was 85.2% and 72.7%, respectively, and there was no difference between living and deceased donors. The graft function of the living donor was better at 1 and 3 months, but was similar from the 6th month onward. There were 6 vascular complications (4 of them vascular thromboses, 1 laceration and 1 renal artery stenosis) and 2 perirenal collections. Seventeen grafts were lost, 6 due to death, 5 with a functioning graft, 5 due to surgical complications, 3 due to chronic rejection and 3 due to recurrence of the original disease. Conclusion: The extraperitoneal access is a valid kidney transplantantion technique in children weighing less than 15 kg, ensuring good patient and graft survival, and an acceptable rate of complications, independent of source of donor, living or deceased, or size of graft, whether from an adult or from a child.

Whilst the major caveat to the success of organ transplantation remains the severe lack of donor organs, rejection is still a primary confounding factor to transplant outcomes. This is an allospecific response that occurs when the recipient immune system recognises conserved proteins on donor-derived cells as 'non-self'. Currently, all immunosuppressive regimes target the recipient immune response, ignoring the large donor immune repertoire despite these cells playing a central role in acute rejection. This is likely as a result of a lack of understanding of the temporal migration of the donor compartment and its contribution to the inflammatory cascade that ensues. The development of ex vivo perfusion provides the opportunity to assess this in isolation, with no confounding factors. Furthermore, inducing the mobilisation of passenger leukocytes on an ex vivo circuit allows their removal prior to transplantation. Reducing the inflammatory burden of donor organs has the potential to impact on the clinical outcome of patients, manifesting as a reduction in the incidence or severity of acute rejection. The aim of this PhD thesis was to characterise the donor immune compartment of lungs and kidneys, to assess the impact of ex vivo perfusion on this, and determine the post-transplant impact of removing a proportion of these cells. For this purpose, donor lungs were perfused using ex vivo lung perfusion (EVLP) and the immune compartment characterised. A comparison of EVLP versus standard transplanted lungs was performed using a porcine transplant model. Clinical parameters were recorded and a histological assessment of cellular infiltration was performed to diagnose the incidence of acute rejection. To determine if these results were translatable to other organs, a porcine model of kidney ex vivo perfusion was established. In both models, a significant efflux of donor leukocytes was observed and inflammatory mediators detected. In a transplant model of EVLP, reducing the transfer of these passenger leukocytes translated into improved clinical outcomes, manifesting as a lower incidence of acute rejection, for animals receiving EVLP lungs compared to a standard transplant. Similar benefit is likely to occur following transplantation of perfused kidneys. This study describes for the first time the contribution of donor organs to the inflammatory processes that ensue following transplantation. It is clear that this untargeted population is of significant importance in clinical outcomes. Immunomodulatory strategies to alter the donor immune environment prior to transplantation therefore warrant development.

La transplantation pulmonaire est une alternative thérapeutique ultime dans de nombreuses maladies pulmonaires sévères, en particulier chez des patients atteints de mucoviscidose (Cystic Fibrosis) , de fibrose pulmonaire idiopathique (IPF idiopathie pulmonary fibrosis), delymphangioléiomyomatose (LAM) ou d'hypertension pulmonaire. Cependant, les besoins en greffe dépassent largement le nombre de transplantations pratiquées en France. Les causes de ce déséquilibre incluent un trop faible nombre de donneurs potentiels en raison de critères actuels pour l'acceptation du greffon qui sont restreints car les biomarqueurs biologiques de qualité et de viabilité du greffon n'ont pas été décrits à ce jour. L'une des possibilités d'augmenter le nombre de donneurs, est de mettre en évidence des biomarqueurs de la qualité du greffon et d'étendre les critères d'acceptabilité du greffon en permettant les prélèvements à partir de donneurs à coeur arrêté est une possibilité. Les prélèvements pulmonaires après arrêt cardiaque sont effectués, en clinique expérimentale chez l'homme, en Espagne depuis trois ans. Il existe dans ce pays une législation très favorable et surtout une conscience collective qui rend le don d'organes extrêmement facilité. Il est donc essentiel d'optimiser l' utilisation de cette ressource. Pour cela, la mise en place de critères de validation de la qualité du greffon est une donnée clé pour palier à ce manque de transplantation pulmonaire. Les critères d'acceptabilité d'un greffon pulmonaires sont basés sur des données cliniques n'existant pas de biomarqueurs biologiques de qualité et de viabilité du greffon. Nous proposons ici l'utilisation de la métabolomique par spectroscopie en Résonance Magnétique Nucléaire à haute résolution par rotation à angle magique (RMN HRMAS) pour mettre en évidence des biomarqueurs de la qualité du greffon. La métabolomique par la spectroscopie RMN HRMAS, est une technique d'analyse rapide (20 minutes) et originale, caractérisée par l'analyse directe d'un tissu biologique intact sans nécessité d'extraction. Cette technique étudie les profils métaboliques dans le but de mettre en évidence des biomarqueurs métaboliques. La métabolomique a été largement utilisée dans des études en cancérologie pour la détermination de la malignité d'un tissu (Bertini 1. et coll. , Canser Res. 2012/ Griffin JL et coll ., Nat Rev Cancer, 2004/ Li M. et coll. , PLoSOne, 2011/ O'connell TM. Bioanalysis. 2012/ Ma Y. et coll. Mol Biol Rep. 2012). Cependant, très peu de publications dans la littérature s'intéressent au domaine de la transplantation et à la qualité du greffon (Rocha C. et coll. , Journal of Proteome Research, 2011/ RobertR. et coll. J Critical Ca re 2010/ Stenlund H. et coll., Chemometrics and Intelligent Laboratory Systems, 2009/ Du a rte F.L. et coll. , Anal.Chem.2005). Ce projet de thèse avait pour objectifs de :1. Etudier la faisabilité d'utiliser la métabolomique par la spectroscopie RMN HRMAS pour évaluer la qualité du greffon pulmonaire.2. Mettre en évidence de potentiels biomarqueurs de la qualité du greffon pulmonaire.3. Evaluer une éventuelle introduction de cette technique en pratique courante dans un environnement hospitalier. Pour répondre à ces objectifs, il a été entrepris :• Etudier les métabolomes pulmonaires de différentes espèces animales, puis de les comparer au métabolome du poumonhumain afin d'identifier le modèle expérimental le plus adapté à la transplantation pulmonaire.• Evaluer la qualité du greffon chez un modèle animal (porc Large White) pour la transplantation pulmonaire (modèle expérimental de préservation pulmonaire en in situ chez le donneur à coeur arrêté, modèle de perfusion pulmonaire en ex vivo sur machine ocs™).• Evaluer l'effet de la perfusion de deux solutions de conservation sur la qualité du greffon pulmonaire chez le porc. / Lung transplantation is a therapeutic alternative in many severe pulmonary diseases, especially in patients suffering from CF (CysticFibrosis), idiopathie pulmonary fibrosis (IPF), lymphangioleiomyomatosis (LAM) or pulmonary hypertension . However, the need fortransplantation far outweighs the number of transplants performed in France. The causes of this imbalance include an insufficient number of potential donors because of the current criteria for the acceptance of the graft that are restricted as biomarkers of quality and viability of the graft have not been described so far.One of the possibilities to increase the donor pool is to identify biomarkers of the quality of the graft and expand the criteria foracceptability of the graft allowing withdrawals from non-heart-beating donors. The lung taking were performed after cardiac arrest inclinical trials carried out on humans for three years in Spain.lt is therefore essential to optimize the use of this resource. For this, the establishment of criteria for validating the quality of the graft isgiven a key to solve this problem of lung transplantation. The criteria for acceptability of a lung transplant are based on clinical data inabsence of biomarkers of quality and viability for the lung graft.We propose the use of the metabolomics by high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HRMASNMR) to highlight potential biomarkers for the quality of the graft.The Metabolomics by NMR HRMAS spectroscopy is an original analytical technique characterized by a rapid analysis (20 minutes)performed on intact biopsy samples without extraction prior to analysis. This technique studies the metabolic profiles in arder to identifymetabolic biomarkers. Metabolomics has been widely used in studies in oncology for the determination of malignancy of a tissue (Bertini et al. , Canser Res. 2012/ Griffin JL et al., Nat Rev Cancer, 2004/ Li M. et al. , PLoS One, 2011/ O'connel! TM. Bioanalysis. 2012/ Ma Y. et al. Mol Biol Rep. 2012). However, very few papers in the literature combine the use of the metabolomics NMR HRMAS and the assessment of the quality of the graft (Rocha C. et al. , Journal of Proteome Research, 2011/ Robert R. et al. J Critical Care 2010/ Stenlund H. et al, Chemometrics and Intelligent Laboratory Systems, 2009/ Duarte F.L. et al. , Anal. Chem. 2005).The purposes of this research work were:1. Studying the feasibility of using the metabolomics by NMR HRMAS for the assessment of the quality of the lung graft2. Assess the metabolome of the lung in degradation conditions and highlight potential biomarkers of the quality of the graft3. Assess the possible use of the metabolomics by NMR HRMAS as a tool in clinical practice within a hospital environment.To answer to these purposes we made experimental experiences as follows:- Studying the lung metabolome of various animal species, and compare them to the human metabolome to identify the most suitableexperimental model for lung transplantation.- Assessing the quality of the graft in an animal model (pig Large White) for lung transplantation (experimental model of lung preservation in situ in the case of non-heart-beating donor, lung model for an ex vivo perfusion using OCS ™ Machine)- Evaluating the effect of perfusion with two preservation solutions on the quality of lung graft in pig model.

Crianças pequenas representam um grupo desafiador no transplante renal. O estudo analisa os resultados, do ponto de vista cirúrgico, do transplante renal em crianças com peso inferior a 15 kg utilizando o acesso cirúrgico extraperitoneal. Métodos: Foram revisados retrospectivamente os prontuários de 62 crianças com peso inferior a 15 kg submetidas a transplante renal entre 1998 e 2010, utilizando o acesso extraperitoneal e anastomose dos vasos renais dos doadores com a aorta ou artéria ilíaca comum e com a veia cava inferior ou ilíaca comum dos receptores. O ureter foi anastomosado à bexiga pela técnica extravesical de Lich- Grégoir. Resultados: Dos 62 transplantes, 32 enxertos (51,6%) eram provenientes de doadores vivos e 30 (48,4%) de doadores falecidos, sendo 28 deles pediátricos. A média de idade no transplante foi de 3,7 ± 2,2 anos (1 a 12), e o peso médio, de 12,3 ± 2,1 kg (5,6 a 14,9), sendo que 10 tinham peso inferior a 10 kg. Em 10 crianças (16,1%) o transplante foi preemptivo e em 5 (8,1%) havia trombose do sistema venoso prévio ao transplante. Em 1 e 5 anos, a sobrevida do paciente foi de 93,2% e 84,2% e a sobrevida do enxerto de 85,2% e 72,7%, respectivamente, sem diferença entre doadores vivos e falecidos. A função do enxerto com doador vivo foi melhor em 1 e 3 meses, mas a partir do 6o mês foi similar. Houve 6 complicações vasculares, sendo 4 tromboses vasculares, 1 laceração e 1 estenose de artéria renal e 2 coleções líquidas. Houve 17 perdas de enxerto, 6 por morte, sendo 5 com enxerto funcionante, 5 por complicações cirúrgicas, 3 por rejeição crônica e 3 por recorrência da doença de base. Conclusão: O acesso extraperitoneal é uma técnica válida no transplante renal de crianças com peso inferior a 15 kg, assegurando boa sobrevida do paciente e do enxerto e aceitável taxa de complicações, independentemente do tipo de doador, se vivo ou falecido, ou do tamanho do enxerto, se de adulto ou de criança. / Small children are a challenging group for kidney transplantation. This study analyzes the results of kidney transplantation in children weighing less than 15 kg using the extraperitoneal surgical access. Methods: A retrospective review of the records of 62 children weighting less than 15 kg was done. The kidney transplantation were performed between 1998 and 2010 using the extraperitoneal access and anastomosis of the renal vessels of donors to the aorta or common iliac artery and to the inferior vena cava or common iliac vein of the recipients. The ureter was anastomosed to the bladder using the Lich-Grégoir extravesical technique. Results: Thirty-two (51.6%) grafts of the 62 transplants were from living donors and 30 (48.4%) from deceased donors, 28 of them pediatric. The mean age at transplantion was 3.7 ± 2.2 years (1 to 12), and the mean weight, 12.3 ± 2.1 kg (5.6 to 14.9), and 10 of them weighed less than 10 kg. In 10 children (16.1%) the transplant was preemptive. Five 5 (8.1%) children presented previous thrombosis of the venous system. At 1 and 5 years, patient survival was 93.2% and 84.2% and graft survival was 85.2% and 72.7%, respectively, and there was no difference between living and deceased donors. The graft function of the living donor was better at 1 and 3 months, but was similar from the 6th month onward. There were 6 vascular complications (4 of them vascular thromboses, 1 laceration and 1 renal artery stenosis) and 2 perirenal collections. Seventeen grafts were lost, 6 due to death, 5 with a functioning graft, 5 due to surgical complications, 3 due to chronic rejection and 3 due to recurrence of the original disease. Conclusion: The extraperitoneal access is a valid kidney transplantantion technique in children weighing less than 15 kg, ensuring good patient and graft survival, and an acceptable rate of complications, independent of source of donor, living or deceased, or size of graft, whether from an adult or from a child.

La pénurie d'organes a amené les équipes de transplantation à élargir l'acceptation des greffons à des organes provenant de donneurs marginaux.Le recours aux greffons marginaux impose de réduire les lésions induites durant l'IR, et doit conduire à une prise en charge optimisée de façon à limiter le risque de PNF et de RRF.C'est face à cette situation, que la question de la perfusion d'organe et de l'utilisation des machines de perfusion s'est posée et a justifié la réalisation de ce travail.Un modèle d'autotransplantation chez le porc a été choisi car il permet d'évaluer les effets de l'IR sur le rein.Les reins ont subi une ischémie contrôlée. La conservation des reins a été randomisée soit en incubation statique dans IGL-1 ou Belzer MPS soit sur machine de perfusion.Nous avons utilisé comme paramètre d'étude : la survie des animaux, différents dosages biologiques, une analyse histologique et une évaluation immunologique par RTqPCR des certains gènes impliqués dans le mécanisme lésionnel d'IR.Nos résultats montrent au total :-Que la machine de perfusion RM3 diminue le risque de RRF et PNF post greffe.-La supériorité de l'IGL-1 sur le Belzer MPS ; il existe un effet propre de la solution IGL-1 pour moduler les mécanismes inflammatoires et immunologiques liés aux lésions d'IR.-Que la reprise de fonction et la PNF dépendent du liquide utilisé et qu'il existe un effet d'addition entre l'IGL-1 et la machine de perfusion. / Organ shortage has led transplant teams to re-evaluate their acceptance criteria and to increase their use of marginal donor organs (ECD and DDAC). For this reason, it is necessary to reduce the lesions due to IR. By optimizing organ conservation, the risk of PNF and RRF can be limited, and organ survival increased. The question of organ perfusion and the use of perfusion machines arose in this context, leading to the work we present here.We chose to evaluate the effects of IR on a porcine auto-graft model, that being the gold standard for the study of IR lesions.The kidneys were subject to hot ischemia for 60 minutes, and then to cold ischemia for 22 hours, during which they were randomly conserved either through static incubation in IGL-1 or Belzer MPS or on a perfusion machine.We studied the animal survival rate, various bioassays, histological analysis and immune reactions though RTqPCR of certain genes involved in IR lesion mechanisms.Our results show:-That the RM3 perfusion machine decreases the RRF and PNF post-graft risk and thus the perfusion machine conservation is better than static conservation.-IGL-1 superiority over Belzer MPS; IGL-1 solution independently modulates inflammatory and immunological mechanisms linked to IR lesions.-That function recovery and PNF depend on the liquid used where there is an additive effect between the use of IGL-1 and the use of a perfusion machine.

Introdução: A técnica de criopreservação de tecido ovariano tem sido vista como tratamento promissor e se apresenta como a principal maneira de preservar a fertilidade em pacientes pré-púberes e em mulheres que necessitam de tratamento do câncer de imediato. Contudo, atualmente, ainda existem obstáculos em relação ao autotransplante de tecido ovariano criopreservado, devido a fatores como lesão isquêmica, assim como danos causados pelo processo durante o congelamento, bem como a escolha do melhor local para o enxerto. Objetivo: Verificar a possível restauração da função ovariana, analisando a histologia do ovário transplantado em ratas adultas estéreis, após transplante autólogo de tecido ovariano criopreservado em fase pré-púbere. Métodos: Foram utilizadas 45 ratas Wistar com 30 dias de idade, que foram divididas aleatoriamente em três grupos: Grupo Controle (n = 15), férteis normais; Sham (n = 15), submetidas à ooforectomia bilateral; Transplante (n = 15), submetidas à ooforectomia bilateral, seguida de transplante autólogo na região dorsal entre as escápulas. A partir do d35, foram realizadas observações quanto à maturidade sexual, através da análise da abertura vaginal e de esfregaços vaginais, para avaliação do ciclo estral. Após observação da fase do ciclo estral, os animais foram eutanasiados. E, amostras de tecidos foram coletadas e processadas para avaliação histológica dos implantes ovarianos; considerando: organização estrutural do tecido transplantado e adjacente, bem como o desenvolvimento folicular. Resultados: Quanto às avaliações de maturidade sexual, através das análises de abertura vaginal e da análise microscópica do material obtido dos esfregaços vaginais, foi possível observar que os animais do Grupo Controle, que eram férteis ciclaram normalmente. As ratas do Grupo Sham e Transplante não apresentaram ciclo regular, permanecendo em diestro. As avaliações histológicas das amostras de tecido de ovário pré-púbere, implantados em fêmeas adulto jovens, evidenciaram degeneração ovariana; uma vez que estes apresentaram fibrose e áreas de necrose, o que provavelmente impossibilitou o desenvolvimento folicular, nas ratas que receberam o transplante. Conclusão: A técnica de transplante de tecido ovariano em ratas é uma técnica relativamente simples de ser executada, e se mostrou eficaz na manutenção do massa corporal dos animais durante o período observado. Este achado sugere que houve produção hormonal, oriunda do ovário transplantado, fato este que encoraja as pesquisas neste sentido, a fim de se obter uma técnica que restaure a produção de folículos viáveis em pacientes estéreis. Apesar de ter apresentando indícios de falência do enxerto e isquemia no tecido transplantado, os resultados preliminares desta investigação precisam ser complementados com estudos adicionais, a fim de buscar as melhores condições para a obtenção de maior eficácia dos transplantes autólogos de tecido ovarianos criopreservados. / Introduction: Ovarian tissue cryopreservation is a promising treatment and it is presented as the main way to preserve fertility in prepubertal patients and women who need cancer treatment immediately. However still remain obstacles related to the ovarian tissue cryopreserved autograft due to ischemic injury, damage caused by the freezing process and selecting the best location for the graft. Objective: Investigate a possible restoration of the ovarian function by analyzing the histology of the ovary transplanted into sterile adult rats after autologous transplantation of ovarian tissue cryopreserved in prepubertal phase. Methods: 45 Wistar rats, 30 days old,which were randomly divided into three groups: control group (n = 15), normal fertile; Sham group (n = 15), underwent bilateral oophorectomy; Transplantation group (n = 15), underwent bilateral oophorectomy followed by autologous transplantation in the scapular area. From the d35, sexual maturity was observed by examining the vaginal opening and vaginal smears, for evaluation of the estrous cycle. After observing the phase of the estrous cycle, the animals were euthanized. The tissue samples were collected and processed for histological evaluation of ovarian implants; where structural organization of the transplanted tissue and adjacent as well as follicular development were analyzed. Results: Regarding sexual maturity evaluations, observed by vaginal opening analysis and microscopic analysis of material obtained from vaginal swabs, we could observe that the animals in the control group cycled normally. The rats of Sham and Transplant Group showed no regular cycle, staying in diestrus phase. The histological assessments of prepubertal ovarian tissue samples implanted in young adult females showed ovarian degeneration, since they had areas of necrosis and fibrosis, which probably impeded the follicular development in these rats. Conclusion: The ovarian tissue transplantation technique in rats is a relatively simple technique, and is effective in body mass maintenance of animals during the observed period. This finding suggests that there were hormone production originated from the transplanted ovaries, and this, encourages research in order to obtain a technique to restore the production of viable follicles in sterile patients. Despite presenting evidence of graft failure and ischemia in the transplanted tissue, the preliminary results of this investigation need to be supplemented with additional studies in order to get the best conditions for achieving greater effectiveness of autologous transplantation of cryopreserved ovarian tissue.

Made available in DSpace on 2014-06-11T19:33:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-07-21Bitstream added on 2014-06-13T20:44:59Z : No. of bitstreams: 1 bonfante_s_dr_araca.pdf: 1718096 bytes, checksum: f842700dfddf9a4f0edab9314172252a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo do presente estudo foi avaliar histológica e histometricamente o padrão de cicatrização de defeitos de deiscência periodontal tratados com enxerto de tecido conjuntivo subepitelial (ETC) ou matriz dérmica acelular (MDA), cada qual comparado ao tratamento com Retalho posicionado Coronal (RPC). Foram utilizados 10 cães, divididos em dois grupo de 5. Defeitos de deiscência óssea (6x8 mm) foram criados nos caninos superiores e os grupos divididos seguindo um modelo de boca dividida de acordo com o tratamento. Em um grupo de 5 cães foi realizado o tratamento com ETC e no lado contralateral RPC e o outro grupo de 5 cães foi tratado com MDA e RPC no lado contralateral. Após 3 meses pós-operatórios os animais foram submetidos à eutanásia e os blocos processados para análise histológica e histométrica. Os parâmetros histométricos avaliados incluíram extensão de tecido epitelial (TE), nova inserção (NITC) e aposição de tecido conjuntivo (ATC), novo cemento (NC) e novo osso (NO). Os dados histométricos transformados em porcentagem, foram estatisticamente analisados pelo teste ANOVA, seguido pelo teste de Tukey... / The aim of the present study was to evaluate histologically and histometrically the healing pattern of periodontal dehiscence defects treated with Subepithelial Connective Tissue Graft (CTG) or Acellular Dermal Matrix (ADM), when compared to the Coronally Positioned Flap (CPF). Dehiscence bone defects (6x8mm) were created bilaterally in the maxillary canine area of dogs (n=10, divided in 2 groups). By means of a split mouth test design, a group of 5 dogs was randomly allocated to receive subepithelial connective tissue graft (CTG) in one of the sides and a coronally positioned flap (CPF) in the opposing side. The remaining 5 dogs had acellular dermal matrix graft (ADM) as a treatment in either one of the sides and CPF on the other side. Three months after surgery, dogs were euthanized and blocks of interest were processed. The histometric parameters for the latter included lenght of epithelial tissue (ET), new attachment (NACT) and connective tissue aposition (CTA), new cementum (NC), and new bone (NB). Histometric data were converted to percentage and statistically analyzed by ANOVA followed by Tukey test at a p<0.05. No significant difference was detected in the following parameters for both groups: ET yielding... (Complete abstract click electronic access below)

Objectif De nombreux travaux ont souligné le rôle de la Cyclosporine A (CsA) dans la prévention des lésions d'ischémie-reperfusion (I/R) mais aucun n'a été effectué sur poumons isolés de grands mammifères. Notre objectif était de mesurer pour la première fois les effets de la CsA sur les lésions d'I/R dans un modèle de poumons porcins reperfusés ex vivo, en évaluant plusieurs doses de CsA pour différents temps d'ischémie. Méthodes L'expérimentation A a été conduite sur 4 groupes de 8 paires de poumons chacune : un groupe contrôle et 3 groupes recevant différentes concentrations de CsA (1, 10 ou 30 μM) au moment de l'ischémie et au début de la reperfusion, après 2 heures d'ischémie. L'expérimentation B a été conduite sur 3 groupes de 5 paires de poumons chacune. Les poumons de chaque paire étaient séparés juste après le début de l'ischémie. Les premiers poumons étaient évalués après une ischémie de 2 heures (jour 0), sans CsA. Les seconds poumons étaient évalués après une ischémie de 24 heures (jour 1), soit sans soit avec CsA (1 ou 5 μM), administrée le cas échéant au début de la reperfusion. Résultats La CsA augmentait le rapport PO2/FiO2 avec un effet dose mais augmentait également la pression artérielle pulmonaire, la pression capillaire et les résistances vasculaires pulmonaires, à 10 et 30 μM mais pas à 1 ni 5 μM. Les poumons qui recevaient 30 μM de CsA affichaient des concentrations élevées en cytokines pro-inflammatoires. La concentration en RAGE (receptor for advanced glycation endproducts) dans le lavage broncho-alvéolaire diminuait avec la CsA à J1 en comparaison à J0. Conclusions Lors de l'I/R pulmonaire, les bénéfices cellulaires des doses élevées de CsA sont contrebalancés par ses effets hémodynamiques sur la microvascularisation. A faibles doses, la CsA semble améliorer la fonction pulmonaire. / Objective Several works highlighted the role of Cyclosporine A (CsA) in the prevention of ischemia reperfusion (I/R) injuries but none on isolated lungs of big mammals. Our objective was to measure for the first time the effects of CsA in I/R injuries in an ex vivo reperfused pig lungs model, by evaluating several doses of CsA for different times of ischemia. Methods Experimentation A was performed on 4 groups of 8 pairs of lungs each: a control group and 3 groups receiving different concentrations of CsA (1, 10 or 30 μM) at the time of ischemia and at the beginning of the reperfusion, after a 2 hours ischemia. Experimentation B was performed on 3 groups of 5 pairs of lungs each. Lungs from each pair were separated just after the beginning of ischemia. The first lungs were evaluated after a 2 hours ischemia (day 0), without CsA. The second lungs were evaluated after a 24 hours ischemia (day 1), either without or with CsA (1 or 5 μM), administered when appropriate at the beginning of the reperfusion. Results CsA improved the PO2/FiO2 ratio with a dose dependent effect but increased pulmonary arterial pressure, capillary pressure, and pulmonary vascular resistances, at 10 and 30 μM but neither at 1 or 5 μM. Lungs receiving 30 μM of CsA displayed elevated concentrations in pro-inflammatory cytokines. Concentrations in RAGE (receptor for advanced glycation endproducts) in broncho-alveolar lavage decreased with CsA at day 1 compared to day 0. Conclusions During pulmonary I/R, the cellular benefits of high doses of CsA are counterbalanced by its hemodynamic effects on microvascularisation. At low doses, CsA seems to improve lung function.

Orientador: Álvaro Francisco Bosco / Banca: Maria José Hitomi Nagata / Banca: Valdir Gouveia Garcia / Banca: Maria Lúcia Rubo de Rezende / Banca: Cristiane Aparecida de Oliveira / Resumo: O objetivo do presente estudo foi avaliar, por meio de análise histológica e histométrica, o processo de reparo de defeitos de furca Classe II, em cães, criados cirurgicamente e tratados com enxerto de osso autógeno em neoformação (OAN) associado ou não a barreira de tecido conjuntivo (TC). Foram utilizados os 3os e 4os pré-molares mandibulares de 6 cães machos para a criação dos defeitos de furca. Inicialmente foram extraídos os 1os molares superiores, com o intuito de criar uma área de rebordo ósseo edêntulo. Após 28 dias, foram confeccionados alvéolos cirúrgicos nestas áreas, com finalidade de obter osso autógeno em neoformação. Decorridos 14 dias, os defeitos de furca classe II, com medidas de 5mm (altura) e 2mm (profundidade), foram criados cirurgicamente e tratados imediatamente. Os dentes foram distribuídos, por meio de randomização bloqueada, em 4 grupos experimentais (n=6): Grupo C (GC): defeito preenchido somente pelo coágulo sanguíneo; Grupo O (GO): preenchimento com OAN; Grupo B (GB): o defeito, preenchido pelo coágulo sanguíneo, foi recoberto por barreira de TC; e Grupo O/B (GO/B): preenchimento com OAN associado a barreira de TC. Os retalhos foram suturados de forma a cobrir totalmente os defeitos. A eutanásia dos animais foi realizada após 90 dias do tratamento dos defeitos de furca criados. Após remoção das peças e tratamento laboratorial de rotina, foram obtidos cortes histológicos seriados no sentido mésio-distal, corados com hematoxilina e eosina ou Tricrômico de Masson. Cinco cortes de cada dente foram selecionados, de forma a representar toda a extensão da furca, para as análises histológica e histométrica. Foram avaliadas medidas lineares e medidas de área da cicatrização periodontal. Os dados obtidos por meio... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The purpose of this study was to histologically and histometrically evaluate the healing of surgically created Class II furcation defects treated using an autogenous bone in neoformation (ABN) graft with or without a connective tissue graft (CT) barrier. The third and fourth mandibular premolars (P3 and P4) of six mongrel dogs were used in this study. Initially, the both first upper molars were extracted, aiming the establishment of an edentulous ridge. After 28 days, surgical alveolus was produced en these areas. After 14 days, Class II furcation defects (5 mm in height and 2 mm in depth) were surgically created and immediately treated. Teeth were randomly divided into four groups: Group C (GC), in which the defect was filled with blood clot only; Group AB (GAB), in which the defect was filled with ABN graft; Group CB (GCB), in which the defect, filled with blood clot, was covered by CT barrier; and Group AB/CB (GAB/CB), in which the defect was filled with ABN graft and covered by a CT barrier. Flaps were repositioned to cover all defects. The animals were euthanized 90 days post-surgery. Vestibule-lingual serial sections were obtained and stained with either hematoxylin and eosin or Masson's trichrome. Histologic and histometric analyses, using a image-analysis software, were performed. Linear and area measurements of periodontal healing were evaluated and calculated as a percentage of the original defect for statistical analysis (analysis of variance; P <0.05). Histologically, the collagen fibers tending to a parallel direction, in groups treated with ABN graft. Although, in the GCB, the periodontal ligament seemed better organized. Anquilosis was observed in their groups that received ABN, whereas the epithelial migration was observed mainly in the specimens of the GC and GCB. The osseous bonefill of the furcation defect was... (Complete abstract click electronic access below) / Doutor

La cirrhose et le carcinome hépatocellulaire liés au virus de l’hépatite C (HCV) sont des indications majeures de transplantation hépatique. La réinfection du greffon par le HCV est systématique et sans moyen de prévention. Les mécanismes par lesquels le virus échappe au système immunitaire afin de réinfecter le greffon sont mal connus. Ce travail de thèse a permis d’identifier de nouveaux mécanismes et des déterminants clés impliqués dans la persistance virale. Nous avons identifié que l’utilisation des facteurs d’entrée cellulaires par le HCV évolue simultanément avec ses capacités d’échappement aux nAbs, et démontré qu’apo E est un composant clé des lipoparticules virales intervenant dans la persistance du HCV. L’identification de nouveaux facteurs viraux et cellulaires impliqués dans l’échappement viral, apporte de nouvelles perspectives dans le développement de nouvelles stratégies de prévention de la réinfection ainsi que dans le développement d’un vaccin prophylactique. / Hepatitis C virus (HCV)-cirrhosis and HCC are leading indications for liver transplantation. Reinfection of the liver graft is universal and no way of prevention exists. Mechanisms employed by HCV to evade the immune system and reinfect the liver graft are unknown. This thesis work identified new mechanisms and key determinants involved in viral persistence. Weidentified a novel clinically and therapeutically important mechanism of viral evasion, where coevolution simultaneously occurs between cellular entry factor use and escape from neutralization. We also discovered that virus-apoE interaction is unique mechanism of viral evasion from neutralizing antibodies. Identification of new viral and host factors involved in viral escape paves the way in the development of new prevention strategies and prophylactic vaccine.