Submitted by PPG Odontologia (odontologia-pg@pucrs.br) on 2018-03-13T18:14:07Z No. of bitstreams: 1 GUILHERME_PIVATTO_LOUZADA_DIS.pdf: 4084164 bytes, checksum: 92327e9e0d6d6a7868235af68dc557f1 (MD5) / Approved for entry into archive by Tatiana Lopes (tatiana.lopes@pucrs.br) on 2018-03-21T11:56:19Z (GMT) No. of bitstreams: 1 GUILHERME_PIVATTO_LOUZADA_DIS.pdf: 4084164 bytes, checksum: 92327e9e0d6d6a7868235af68dc557f1 (MD5) / Made available in DSpace on 2018-03-21T12:03:36Z (GMT). No. of bitstreams: 1 GUILHERME_PIVATTO_LOUZADA_DIS.pdf: 4084164 bytes, checksum: 92327e9e0d6d6a7868235af68dc557f1 (MD5) Previous issue date: 2018-01-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Objective: The present study aimed to evaluate the effects of systemic ozonotherapy on bone remodeling of critical defects in the calotte of rats, with and without a xenograft presence, using an animal model of immunosuppression induced by corticosteroids. Methods: Sixty male Wistar rats (180-220 g), distributed in 8 experimental groups (N = 8 / group) were used. For corticosteroid therapy (groups II, IV, VI and VIII), the animals received dexamethasone (1 mg / kg; i.p.), one injection every 48 h, starting two weeks before the surgical procedures, extending to euthanasia. Control animals (groups I, III, V and VII) received saline solution (10 ml / kg, i.p.) at the same time intervals. After two weeks of corticosteroid therapy, the animals were anesthetized with ketamine and xylazine (100 and 10 mg / kg, i.p.) to produce two critical defects in the parietal bones of the skull cap (5 mm diameter). The defects were filled with clot (groups I, II, V and VI) or Bio-Oss x xenograft (Geistlish Biomaterials, Germany) (groups III, IV, VII and VIII). In all groups, the right defect was covered by a Bio-Gide? collagen membrane (Geistlish Biomaterials, Germany). For systemic ozono-therapy, animals of groups V, VI, VII and VIII received an application of ozone (0.7 mg / kg, i.p.) every day for 7 days, starting immediately after surgery. Four weeks after the defects were created, the animals were euthanized, and the skull caps were collected for histological evaluation of the bone neoformation with hematoxylin and eosin (HE) staining. The protocols were approved by the Ethics Committee on the Use of Animals (7691). Qualitative histological analyzes were performed based on the pattern of connective tissue formation around the defect, existence of inflammatory cells in the region, aspect of bone trabeculated, osteoblastic activity around the bone matrix and existence of graft particles in the region. Results: Groups V and VII presented greater bone areas along the surface of the defect compared to VI and VIII, because of the potential effects of ozone on bone remodeling, minimizing negative interference in the glucocorticoid bone microarray. The presence of the xenografts groups III, IV, VII, VIII, provided the covering of an extensive area of the defect, forming a more prominent immature bone matrix near the edges of the defects, with the presence of graft particles dispersed in the central area. The presence of the type I collagen membrane was shown to be an important tool in the acceleration of bone remodeling used in all experimental groups, in which, in comparison to contralateral defects in which the membrane was not used, a significant increase of bone just graduated. At the systemic level, the results of ozone treatment allowed the reduction of adverse effects of glucocorticoids, such as the reduction of the body weight of the animals and atrophy of lymphatic organs such as spleen, observed in the groups treated alone with dexamethasone. Conclusion: Given the limitations of the animal study, the use of systemic ozone associated with the xenograft stimulates bone remodeling in critical defects of immunosuppressed rats. / Objetivo: O presente estudo teve por objetivo avaliar os efeitos da ozonioterapia sist?mica sobre o remodelamento ?sseo de defeitos cr?ticos na calota de ratos, com e sem a presen?a de xenoenxerto, utilizando um modelo animal de imunossupress?o induzido por corticoterapia. M?todos: Foram utilizados 64 ratos machos Wistar (180-220 g), distribu?dos em 8 grupos experimentais (N = 8/grupo). Para a corticoterapia (grupos II, IV, VI e VIII), os animais receberam dexametasona (1 mg/kg; i.p.), uma inje??o a cada 48 h, com in?cio duas semanas antes dos procedimentos cir?rgicos, se estendendo at? a eutan?sia. Os animais controles (grupos I, III, V e VII) receberam solu??o salina (10 ml/kg; i.p.), nos mesmos intervalos de tempo. Ap?s duas semanas do in?cio da corticoterapia, os animais foram anestesiados com quetamina e xilazina (100 e 10 mg/kg; i.p.), para a confec??o de dois defeitos cr?ticos nos ossos parietais da calota craniana (5 mm de di?metro). Os defeitos foram preenchidos com co?gulo (grupos I, II, V e VI) ou com xenoenxerto Bio-Oss? (Geistlish Biomaterials, Germany) (grupos III, IV, VII e VIII). Em todos os grupos, o defeito da direita foi coberto por uma membrana de col?geno Bio-Gide? (Geistlish Biomaterials, Germany). Para a ozonioterapia sist?mica, os animais dos grupos V, VI, VII e VIII receberam uma aplica??o de oz?nio (0,7 mg/kg; i.p.), todos os dias, durante 7 dias, iniciando imediatamente ap?s a cirurgia. Decorridos quatro semanas da cria??o dos defeitos, os animais foram eutanasiados e as calotas cranianas foram coletadas para avalia??o histol?gica da neoforma??o ?ssea, com colora??o de hematoxilina e eosina (HE). Os protocolos foram aprovados pela Comiss?o de ?tica no Uso de Animais (7691). Foram realizadas an?lises histol?gicas qualitativas baseado no aspecto do trabeculado ?sseo, atividade osteobl?stica em torno da matriz ?ssea, padr?o de forma??o de tecido conjuntivo na ?rea do defeito, e exist?ncia de part?culas de enxerto na regi?o. Resultados: Os grupos V, VII apresentaram maiores ?reas ?sseas ao longo da superf?cie do defeito, em compara??o com os VI e VIII, como consequ?ncia dos efeitos potenciais do oz?nio no remodelamento ?sseo, minimizando as interfer?ncias negativas na microarquitetura ?ssea causada pelos glicocortic?ides. A presen?a do xenoenxerto nos grupos III, IV, VII, VIII, proporcionou o recobrimento de extensa ?rea do defeito, formando matriz ?ssea imatura mais proeminente nas proximidades dos bordos dos defeitos, com presen?a de part?culas de enxerto dispersas na ?rea central. A presen?a da membrana de col?geno tipo I, se mostrou um importante recurso na acelera??o do remodelamento ?sseo utilizado em todos os grupos experimentais, na qual em compara??o com os defeitos contralaterais em que n?o foi utilizado a membrana, pode ser visto um significativo aumento de osso rec?m-formado. A n?vel sist?mico, os resultados do tratamento com oz?nio possibilitaram a diminui??o de efeitos adversos dos glicocorticoides, tais como a diminui??o do peso corporal dos animais e atrofia de ?rg?os linf?ticos como o ba?o, observados nos grupos tratados isoladamente com a dexametasona. Conclus?o: Diante das limita??es do estudo animal, a utiliza??o de oz?nio sist?mico associado ao xenoenxerto, estimula o remodelamento ?sseo em defeitos cr?ticos de ratos imunossuprimidos.

Made available in DSpace on 2014-12-17T15:32:18Z (GMT). No. of bitstreams: 1 EmelineNAL.pdf: 2548007 bytes, checksum: d30f88fa3814f13a0bc3ee25b49d05fb (MD5) Previous issue date: 2010-02-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Bone marrow transplantation (BMT) is currently the best therapeutic option for patients with hematologic diseases, solid tumors or autoimmune disorders. It is characterized by intravenous infusion of hematopoietic stem cells in order to restore marrow function. However, this procedure requires concomitant immunosuppression treatment, which favors the development of certain complications, often manifested in the oral cavity. This study aimed to evaluate the incidence of oral changes in patients undergoing BMT and to correlate these results with clinical aspects related to the patients and the transplants performed. This is a prevalence study, with cross-sectional design, carried out in a BMT service at the Institute of Onco-Hematology of Natal (ION) and Natal Hospital Center. Data collection was based on questionnaires, clinical examination of the oral cavity and consultation in the medical records. The sample consisted of 51 patients undergoing BMT. After the analysis, was found a general status with good health conditions and presence of oral changes in about half of patients who composed the sample. The manifestations observed were, in decreasing order of frequency: mucositis; gingival alteration and thrombocytopenic purpura; mucosal pigmentation; lichenoid reaction and candidiasis. The oral changes were observed more frequently in cases of allogeneic TMO, in different periods post-transplant, without significant differences related to the source of cells. It was found statistically significant association between the presence of graft-versus-host disease (GVHD) and oral changes (p < 0,001). Therefore, it is concluded that there is a relatively high incidence of changes in oral cavity of patients receiving bone marrow transplantation, a fact which confirms the need to consider this site for examination, diagnosis, treatment and prognosis of possible complications of BMT / O transplante de medula ?ssea (TMO) atualmente constitui a melhor op??o terap?utica para pacientes com doen?as hematol?gicas, tumores s?lidos ou desordens autoimunes. Caracteriza-se pela infus?o intravenosa de c?lulas progenitoras hematopo?ticas com o objetivo de restabelecer a fun??o medular. No entanto, esse procedimento requer tratamento concomitante de imunossupress?o, o que favorece o desenvolvimento de determinadas complica??es, as quais freq?entemente se manifestam na cavidade oral. Este estudo objetivou avaliar a incid?ncia de altera??es orais em pacientes submetidos ao TMO e correlacionar esses resultados com aspectos cl?nicos referentes aos pacientes e aos transplantes realizados. Trata-se de um estudo de preval?ncia, com desenho do tipo seccional, realizado no servi?o de TMO do Instituto de Onco-Hematologia de Natal (ION) e Natal Hospital Center. A coleta de dados baseou-se em aplica??o de question?rio, exame cl?nico da cavidade oral e consulta de informa??es nos prontu?rios m?dicos. A amostra foi constitu?da por 51 pacientes submetidos ao TMO. Ap?s a an?lise, constatou-se quadro geral com boas condi??es de sa?de e presen?a de altera??es orais em aproximadamente metade dos pacientes que compunham a amostra. As manifesta??es observadas foram, em ordem decrescente de frequ?ncia: mucosite; altera??o gengival e p?rpura trombocitop?nica; pigmenta??o da mucosa; rea??o liquen?ide e candid?ase. As altera??es orais foram mais frequentes em casos de TMO alog?nico, em diferentes per?odos p?s-transplantes, sem diferen?a significativa quanto ? origem das c?lulas. Constatou-se associa??o estatisticamente significante entre a presen?a de doen?a do enxerto contra hospedeiro (DECH) e altera??es orais (p < 0,001). Portanto, conclui-se que h? uma incid?ncia relativamente alta de altera??es na cavidade oral de pacientes transplantados de medula ?ssea, fato que confirma a necessidade de se considerar a import?ncia desse s?tio para exame, diagn?stico, tratamento e progn?stico de poss?veis complica??es do TMO

Made available in DSpace on 2015-04-14T14:51:18Z (GMT). No. of bitstreams: 1 438527.pdf: 10657738 bytes, checksum: caad3abdc5d9b9863dbe5c3d7120700f (MD5) Previous issue date: 2012-03-26 / Transplantation of solid organs has emerged as a viable therapeutic modality for the treatment of a variety of disorders. Rejection of solid organ allografts is the result of a complex range of interactions involving coordination between both the innate and adaptive immune system. Therewith, a major goal of clinical organ transplantation is to induce a donor-specific unresponsive state in a mature immune system that is free from long-term immunosuppression and chronic rejection. The limitations in the establishment of immunossupressive stratagies led us to search new methods to the modulation of the homeostatic mechanisms that limit and prevent inflammatory responses in allograft tissue. The heat shock protein 70 (Hsp70) has a protective and antiinflamatory role in several animals models like arthritis, colitis, pulmonary fibrosis and brain injury. This protein can modulates both the innate and adaptative immune system. Our group demonstrated that Mycobacterium tuberculosis Hsp70 (Mt Hsp70) can inhibit bone marrow dendritic cells (BMDCs) maturation; however the mechanisms involved in this process has not been completely elucidated. In the present work, we demonstrated that Mt Hsp70 inhibited the acute rejection in two allograft models (a tumor model and a skin allograft model).In both models, we observed an involvement of Tregs. In addition, s.c. Mt Hsp70 injection leds to an increase in Treg population and IL-10 production in the draining lymph node. We also observed that the inhibition of acute rejection induced by Mt Hsp70 was dependent on the presence of toll-like receptor (TLR) 2 in the allograft, and not in the host. In BMDCs, we demonstrated that IL-10 production induced by Mt Hsp70 is dependent on TLR2. Also, we analyzed the phosphorylation of ERK, p38 and Akt after Mt Hsp70 stimulus. We observed an increase in p-ERK expression, but no difference in p-38 and p-Akt levels. The inhibition of ERK abolished the IL-10 production induced by Mt Hsp70. We propose that Mt Hsp70 effect on DCs can be used as a therapeutic approach in transplantation models. / O transplante de ?rg?os s?lidos emergiu como uma terapia vi?vel para o tratamento de uma variedade de patologias. A rejei??o dos enxertos ? resultado de uma s?rie complexa e coordenada de intera??es envolvendo o sistema imune inato e adaptativo. Com isso, o maior desafio no transplante de ?rg?os s?lidos ? induzir um estado irresponsivo e espec?fico ao doador em um sistema imune maduro sem que haja uma imunossupress?o sist?mica e de longo prazo, tudo isso livre de rejei??o cr?nica. As limita??es no estabelecimento de estrat?gias imunossupressoras nos levaram a buscar novos m?todos para a modula??o dos mecanismos homeost?ticos que previnem e limitam as respostas inflamat?rias nos tecidos enxertados. A prote?na de choque t?rmico (Heat shock protein Hsp) 70 tem um papel antiinflamat?rio e protetor em modelos animais experimentais como artrite, colite, fibrose pulmonar e danos cerebrais. Essa prot?ina pode modular tanto o sistema imune inato quanto o adaptativo. Nosso grupo demonstrou que a Hsp70 de Mycobacterium tuberculosis (Mt Hsp70) pode inibir a matura??o de c?lulas dendr?ticas diferenciadas da medula ?ssea (bone marrow dendritic cells BMDCs), por?m o mecanismo envolvido nesse processo ainda n?o foi totalmente esclarecido. Nesse trabalho, demonstramos que a Mt Hsp70 foi capaz de aumentar a sobrevida do enxerto em dois modelos murinos de transplantes (um modelo tumoral e um modelo de aloenxerto cut?neo).Em ambos os modelos, observamos o envolvimento de Tregs. Demonstramos que a administra??o s.c. da Mt Hsp70 levou a um aumento dessas c?lulas nos linfonodos drenantes, al?m de um aumento na produ??o de IL-10. Tamb?m observamos que a inibi??o da rejei??o aguda induzida pela Mt Hsp70 no modelo de aloenxerto cut?neo ? dependente da presen?a do receptor do tipo toll (toll like receptor TLR) 2 no enxerto, e n?o no receptor. Nas BMDCs, vimos que a indu??o da IL-10 induzida pela Mt Hsp70 ? dependente de TLR2. Ainda nessas c?lulas, analisamos a fosforila??o de mol?culas como a ERK, p38 e Akt ap?s o est?mulo com a Mt Hsp70. Observamos um aumento na express?o de p-ERK e nenhuma altera??o nos n?veis de p-p38 e p-Akt. A inibi??o da ERK aboliu a produ??o de IL-10 induzida pela Mt Hsp70. Propomos que esse efeito da Mt Hsp70 sobre as DCs pode servir como interven??o terap?utica em modelos de transplantes.