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Gerluza Aparecida Borges Silvahttp://lattes.cnpq.br/0626838761962488Erika Cristina JorgeAmanda Maria Sena ReisFernando Antônio Mauad de Abreuhttp://lattes.cnpq.br/3587263861317914Igor Daniel Garcia Reis2021-03-25T11:09:26Z2021-03-25T11:09:26Z2018-02-19http://hdl.handle.net/1843/35393A utilização de xenoenxertos derivados de osso bovino tem se configurado como uma alternativa aos autoenxertos na substituição de perdas óssea. Entretanto, sua propriedade de osteoindução e potencial osteogênico ainda são limitados. Por essa razão, algumas estratégias como a desmineralização de fragmentos ósseos, bem como a utilização de células tronco mesenquimais tem sido proposta para a otimização do reparo ósseo. Nesse contexto, este estudo avaliou os efeitos da associação de uma matriz óssea de origem bovina, marca comercial- Criteria®, desmineralizada em EDTA 10%, às CTPD’s (modificadas para eGFP - enhancedgreenfluorescentprotein) no reparo de um defeito ósseo intrabucal em ratos. Os defeitos foram preenchidos por: 1. Matriz óssea desmineralizada (MOD); 2. CTPD’s em suspensão; 3. CTPD+MOD; 4. Coágulo sanguíneo (controle). Os animais foram sacrificados após 7, 14 e 49 dias para avaliações histomorfométricas e imunohistoquímica para GFP (visando o monitoramento das células tronco transplantadas); e após 3 e 7 dias para análise de expressão de genes indicativos de diferenciação osteoblástica (Runx2, Osx e iBSP) por RT-PCRq. Aos 7 dias, o fechamento epitelial foi incompleto em todos os grupos. A morfometria demonstrou que a deposição óssea na área do defeito cirúrgico foi maior no grupo que recebeu a associação proposta, seguido pela enxertia com as CTPD’s isoladas. Este resultado está em concordância com a análise molecular, que demonstrou a expressão do marcador de diferenciação osteoblástica iBSP, 64 vezes maior no mesmo período. Esse padrão de deposição óssea na análise morfométrica se manteve nos períodos de 14 e 49 dias, evidenciando a associação benéfica entre CTPD+MOD para terapias de regeneração tecidual óssea. O nível de maturação da matriz foi avaliado por coloração de PicroSirius Red e revelou a maior maturação no grupo CTPD+MOD. Resumidamente, os resultados demonstraram otimização do processo de regeneração óssea com a associação CTPD+MOD em todos os períodos. O estudo traz esclarecimentos sobre o xenoenxerto de fabricação nacional e apresenta uma proposta de otimização para o seu uso no desenvolvimento de terapias de regeneração tecidual.The use of xenografts derived from bovine bone has been configured as an alternative to autografts in the replacement of bone loss. However, its property of osteoinduction and osteogenic potential are still limited. For this reason, some strategies such as demineralization of bone fragments as well as the use of mesenchymal stem cells have been proposed for the optimization of bone repair. In this context, this study evaluated the effects of the association of a bone matrix of bovine origin, commercial brand - Criteria ®, demineralized in EDTA 10%, to the CTPDs (modified for eGFP - enhancedgreenfluorescentprotein) in the repair of an intrabuccal bone defect in rats. The defects were filled by: 1. Demineralized bone matrix (MOD); 2. TDPs in suspension; 3. CTPD + MOD; 4. Blood clot (control). The animals were sacrificed after 7, 14 and 49 days for histomorphometric and immunohistochemical evaluations for GFP (aiming at the monitoring of transplanted stem cells); and after 3 and 7 days for expression analysis of genes indicative of osteoblastic differentiation (Runx2, Osx and iBSP) by RT-PCRq. At 7 days, epithelial closure was incomplete in all groups. Morphometry demonstrated that bone deposition in the area of the surgical defect was higher in the group that received the proposed association, followed by grafting with the isolated TCDC's. This result is in agreement with the molecular analysis, which demonstrated the expression of the osteoblast differentiation marker iBSP, 64 times higher in the same period. This pattern of bone deposition in the morphometric analysis was maintained in the periods of 14 and 49 days, evidencing the beneficial association between CTPD + MOD for tissue regeneration therapies. The maturation level of the matrix was evaluated by staining of PicroSirius Red and revealed the highest maturation in the CTPD + MOD group. Briefly, the results demonstrated optimization of the bone regeneration process with the association CTPD + MOD in all periods. The study brings clarifications about the xenograft of national manufacture and presents an optimization proposal for its use in the development of tissue regeneration therapies.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Biologia CelularUFMGBrasilICB - INSTITUTO DE CIÊNCIAS BIOLOGICAShttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessXenoenxertosCélulas-tronco mesenquimaisOsteogêneseXenoenxertosCélulas tronco mesenquimaisOsteogêneseOsteoinduçãoDefeito ósseo intrabucalAssociação de células tronco da polpa dentária a enxertos ósseos desmineralizados para otimização de reparos ósseosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALDissertação Igor Reis.pdfDissertação Igor Reis.pdfapplication/pdf15587474https://repositorio.ufmg.br/bitstream/1843/35393/1/Disserta%c3%a7%c3%a3o%20Igor%20Reis.pdf0683c577c1990f9fb0c2f795799ae97aMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/35393/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82119https://repositorio.ufmg.br/bitstream/1843/35393/3/license.txt34badce4be7e31e3adb4575ae96af679MD531843/353932021-03-25 08:09:26.297oai:repositorio.ufmg.br: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Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2021-03-25T11:09:26Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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