Compara??o de diferentes metodologias para descelulariza??o de veias humanas

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Bibliographic Details
Main Author: Oliveira , Mada? Cruz Lopes de
Other Authors: Silva , Jefferson Luis Braga da
Format: Others
Language:Portuguese
Published: Pontif?cia Universidade Cat?lica do Rio Grande do Sul 2015
Subjects:
Online Access:http://tede2.pucrs.br/tede2/handle/tede/6203
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Summary:Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-07-07T12:57:05Z No. of bitstreams: 1 471707 - Texto Completo.pdf: 875696 bytes, checksum: 681f03e704120fe92558e48e363af3d4 (MD5) === Made available in DSpace on 2015-07-07T12:57:05Z (GMT). No. of bitstreams: 1 471707 - Texto Completo.pdf: 875696 bytes, checksum: 681f03e704120fe92558e48e363af3d4 (MD5) Previous issue date: 2015-03-27 === Introduction : Non-living tissues have shown to be an important source of biomaterials, with advantages such as hydrophilicity, low toxicicidade, low immunogenicity, in addition to good adherence and cell multiplication. However, for these biomaterials to be used, techniques of descellularization is being used to make it less antigenic and also to serve as scaffolds.Objectives : The present study aims to compare different agents of descellularization regarding the above mentioned parameters, using detergents known to be effective for these processes.Materials and Methods : The samples used in this study were obtained from saphenous veins of seven individuals donors, in treatment of myocardial revascularization surgery and provided by the team of cardiovascular surgery of Hospital S?o Lucas da PUCRS, after signing the informed consent form.These samples were tested simultaneously three different detergents, sodium dodecyl sulfate (SDS), TRITON-X and trypsin, in addition to a control group containing Phosphate-Buffered Saline (PBS). After the process of descellularization and prior to the determination of cellularity, the samples were fixed in buffered formalin 10% during 24 hours, included in paraffin, sectioned (5 ?) and stained with hematoxylin and eosin. The number of cores was quantified in a radius of 0.025 mm (objective 40x), in duplicate, corresponding to eight fields, with the aid of the software Image Pr?-Plus 4.5.1. Data were analyzed using a model of analysis of variance for repeated measures (randomized blocks) with adjustment for multiple comparisons procedure of Finner's procedure were used.Results : The results evaluated all treatments showed a decrease in the number of cells, and the SDS the agent with greater descellularization. The TRITON-X and the Trypsin obtained similar values and there was no difference between them, for this process.Conclusion : In this way, it could be demonstrated that all detergents used, and the methodology it self, acts as agentes of descellularization in human veins and proved to be a promising agents to remove cells. === Introdu??o : Tecidos n?o vivos tem se mostrado uma importante fonte de biomateriais, apresentando vantagens como hidrofilidade, baixa toxicicidade, baixa imunogenicidade, al?m de boa ades?o e multiplica??o celular. Contudo, para que esses biomateriais sejam utilizados, t?cnicas de descelulariza??o v?m sendo utilizadas para que sejam menos antig?nicos e tamb?m para que sirvam como scaffolds.Objetivos : O presente estudo tem o objetivo de comparar diferentes agentes de descelulariza??o quanto aos par?metros citados acima, utilizando detergentes reconhecidamente eficazes para esses processos.Materiais e M?todos : As amostras utilizadas neste estudo foram obtidas de veias safenas de sete indiv?duos doadores, em tratamento de cirurgia de revasculariza??o do mioc?rdio e fornecidas pela equipe de cirurgia cardiovascular do Hospital S?o Lucas da PUCRS, ap?s assinatura do termo de consentimento livre e esclarecido (TCLE). Nessas amostras foram testados simultaneamente tr?s diferentes detergentes, s?dio dodecil sulfato (SDS), TRITON-X e tripsina, al?m de um grupo controle, contendo Phosphate-Buffered Saline (PBS) e um controle ?a fresco?. Ap?s o processo de descelulariza??o e previamente ? determina??o da celularidade, as amostras foram fixadas em formalina tamponada 10% durante 24horas, inclu?das em parafinas, seccionadas (5?) e coradas com hematoxilina e eosina. O n?mero de n?cleos presentes foi quantificado em um raio de 0,025 mm (objetiva 40x), em duplicata, correspondente a oito campos, com o aux?lio do software Image Pr?-Plus 4.5.1. Os dados foram analisados em modelo de an?lise de vari?ncia de medidas repetidas (blocos casualizados) com ajuste de compara??es m?ltiplas pelo procedimento de Finner.Resultados : Dos resultados avaliados todos os tratamentos mostraram um decr?scimo no n?mero de c?lulas, sendo o SDS o agente com maior descelulariza??o. O TRITON-X e a Tripsina obtiveram valores similares e n?o houve diferen?a entre eles, para esse processo.Conclus?o : Dessa forma, demonstrou-se que todos os detergentes empregados agem como descelularizadores em veias humanas, e mostram-se como promissores agentes de remo??o celular.