Efeitos da membrana amniótica homógena associada ao laser arseneto de gálio-alumínio (λ=660) na reparação de mucosite oral em ratos tratados com 5-fluoruracila: análise histológica e imunohistoquímica
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Data
2011-07-14
Autores
Lima, Gabriela de Morais Gouvêa [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A presente dissertação objetivou a análise dos efeitos da membrana amniótica homógena associada ao laser arseneto de gálio-alumínio sobre a mucosite oral induzida em ratos sob uso do quimioterápico 5-fluoruracila (5-FU). Foram estudados 100 ratos Wistar, divididos em 5 grupos (20 ratos cada grupo); sendo um grupo controle, um grupo tratado com 5- fluoruracila, grupo tratado com 5-FU+membrana amniótica homógena (MAH), grupo 5-FU+laser arseneto de gálio-alumínio, grupo 5- FU+MAH+laser arseneto de gálio-alumínio.Cada grupo foi subdividido em 4 para sacrifício e obtenção dos cortes histológicos, sendo analisados os períodos de 3, 7, 14 e 21 dias após indução da mucosite. Os cortes histológicos foram corados com hematoxilina-eosina para a análise histomorfológica e histomorfométrica. Para a análise imunohistoquímica foi usado o método indireto com sistema de amplificação (Peroxidase, Biotina e Estreptavidina) e as lâminas foram incubadas nos anticorpos primários anti-rato: CD-4, CD-8, VEGF e PCNA e anticorpo secundário LSAB+Sys-HRP. A contra coloração foi feita com Hematoxilina de Mayer. As células imunomarcadas foram contadas na região mediana da lâmina própria, em uma área de aproximadamente 2.41 mm2, usando o programa Axiovision 4.7. Os resultados da histomorfometria foram avaliados de forma descritiva, enquanto para a análise imunohistoquímica usou-se análise de variância (One Way ANOVA) e ao teste de Tukey (GraphPad Prisma 5 for Windows , GraphPad Software, San Diego, California, USA). O nível de significância adotada foi de p< 0.05. Concluiu-se que a MAH apresentou biocompatibilidade, acelerando o processo de reparação das mucosites e que a laserterapia teve efeito antiinflamatório, principalmente nas fases iniciais da mucosite
The objective of this dissertation was to analise the effects of association between amniotic homogen membrane (AHM) and gallium-aluminum arsenide laser on 5-fluoruracil (5-FU) induced oral mucositis healing process in rats. 100 Wistar rats were divided in 5 groups (20 rats each group): control group, group 5-FU (treated with 5- fluoruracil), 5FU+AHM group (trated with 5-FU+AHM), Laser group (treated wuith 5-FU+laser) and 5-FU+MAH+Laser group (trated with 5FU+AHM+laser). Each group were subdivided in fourth according to sacrifice period (3, 7, 14 and 21 days), after mucositis induction. Histological slices were stained with hematoxilin-eosine for histomological analisis. For immunohistochemical and histomorfometric analisis, the indirect method with amplification system (Peroxidase, Biotin and Streptavidin) were used and glasses were incubated with anti-rat antibodies CD-4, CD-8, VEGF and PCNA and secondary antibody LSAB+Sys-HRP. Mayer Hematoxilin was used as couterstain and immunostained cells were counted on middle region of lamina propria, with 2.41 mm2 using Axiovision 4.7 Imaging System. Histomorfometry results were decribed and immunohistochemistry were analised with ANOVA one-way and Tukey test (GraphPad Prisma 5 for Windows, GraphPad Software, San Diego, Califórnia, USA). Significance level was p< 0.05. AHM presented biocompatibility acelerating mucositis healing process and lasertherapy had anti-inflamatory effect, specially on begining phases of healing process
The objective of this dissertation was to analise the effects of association between amniotic homogen membrane (AHM) and gallium-aluminum arsenide laser on 5-fluoruracil (5-FU) induced oral mucositis healing process in rats. 100 Wistar rats were divided in 5 groups (20 rats each group): control group, group 5-FU (treated with 5- fluoruracil), 5FU+AHM group (trated with 5-FU+AHM), Laser group (treated wuith 5-FU+laser) and 5-FU+MAH+Laser group (trated with 5FU+AHM+laser). Each group were subdivided in fourth according to sacrifice period (3, 7, 14 and 21 days), after mucositis induction. Histological slices were stained with hematoxilin-eosine for histomological analisis. For immunohistochemical and histomorfometric analisis, the indirect method with amplification system (Peroxidase, Biotin and Streptavidin) were used and glasses were incubated with anti-rat antibodies CD-4, CD-8, VEGF and PCNA and secondary antibody LSAB+Sys-HRP. Mayer Hematoxilin was used as couterstain and immunostained cells were counted on middle region of lamina propria, with 2.41 mm2 using Axiovision 4.7 Imaging System. Histomorfometry results were decribed and immunohistochemistry were analised with ANOVA one-way and Tukey test (GraphPad Prisma 5 for Windows, GraphPad Software, San Diego, Califórnia, USA). Significance level was p< 0.05. AHM presented biocompatibility acelerating mucositis healing process and lasertherapy had anti-inflamatory effect, specially on begining phases of healing process
Descrição
Palavras-chave
Quimioterapia, Membrana amniótica homógena, Mucosite oral, Oral mucositis
Como citar
LIMA, Gabriela de Morais Gouvêa. Efeitos da membrana amniótica homógena associada ao laser arseneto de gálio-alumínio (λ=660) na reparação de mucosite oral em ratos tratados com 5-fluoruracila: análise histológica e imunohistoquímica. 2011. 94 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Odontologia de São José dos Campos, 2011.