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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2011.tde-30052012-092333
Document
Author
Full name
Eric Marcel Guzovsky
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Vassao, Ruth Camargo (President)
Dagli, Maria Lucia Zaidan
Garcia, Primavera Borelli
Title in Portuguese
Análise de alterações histopatológicas e celulares em tecidos de camundongos C57BL/6J inoculados com células de melanoma experimental B16F10 por via subcultanea e tratados com o imunomodulador Imiquimod 5%.
Keywords in Portuguese
Adjuvantes imunológico
Camundongos
Células cultivada de tumor
Histopatologia animal
Imidazoquinolina
Imunomodulação
Abstract in Portuguese
Imiquimod é uma imidazoquinolina, agonista do TLR-7, ativador de células dendríticas (DC) e indutor da transcrição de diversas citocinas. O objetivo deste estudo é avaliar o crescimento tumoral, a formação de metástases em órgãos internos, a atividade do sistema imune e o tipo de morte celular induzido devido ao tratamento tópico. Camundongos foram inoculados com células de melanoma murino B16F10. O fármaco, na dose de 5mg, foi aplicado no local da inoculação uma vez por dia até o fim do experimento. Foram estudados quatro grupos experimentais, dois inoculado com células tumorais, sendo um tratado e outro não, e outros dois grupos, um recebeu somente o tratamento com o fármaco e o outro não recebeu tratamento. Os animais foram pesados e seus tumores medidos periodicamente. Exemplares dos grupos foram sacrificados em quatro tempos diferentes, imediatamente submetidos à necropsia para a análise de massas tumorais por citometria de fluxo com caspase-3 e anexina-v para investigar a indução de apoptose e/ ou necrose tumoral, e remoção do tumor, pele, Baço, linfonodo e pulmão, que foram fixados, inclusos em parafina, cortados e corados com HE, para análise histológica.
Title in English
Histopathologic analysis of murine C57BL/6J tissues and cells innoculated with B16F10 melanoma cells subcutaneously, treated with the immunemodulator Imiquimod 5%.
Keywords in English
Animal histopathology
Cultured tumor cells
Imidazoquinolina
Immune adjuvants
Immunomodulation
Mice
Abstract in English
Imiquimod is a TLR-7 agonist and activator of dendritic cells (DC), and induces transcription of several cytokines. This study`s objective is to evaluate tumor growth, development of metastasis within internal organs and the immune system`s function, due to the topical treatment. Two groups of C57BL/6J mice were inoculated with B16F10 melanoma cells subcutaneously, and only one of the groups was treated, applied topically at the injection site once a day, followed by a second round of two groups, one treated and the other as a healthy group. Every two tumors and weight were measured. In order to observe the tumor progression and treatment effects, groups of animals were sacrificed on four days apart and immediately submitted to necropsy for a tumor analysis by flow cytometry, to evaluate the induction of apoptosis and necrosis, and to macroscopically observe the tumor development within internal organs, vascularization and posterior inclusion of the organs in paraffin for histological analysis. Tumor, skin, spleen, limph node and lung were colored with HE for histological analysis of migrating immune system cells and tumoral cells in the organs and describing its integrity.
 
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Publishing Date
2012-06-27
 
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