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Master's Dissertation
DOI
https://doi.org/10.11606/D.82.2002.tde-07012003-100025
Document
Author
Full name
Paulo de Tarso Camillo de Carvalho
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2001
Supervisor
Committee
Mazzer, Nilton (President)
Barbieri, Claudio Henrique
Fazan, Valeria Paula Sassoli
Title in Portuguese
Análise da cicatrização de lesões cutâneas através da espectrofotometria: estudo experimental em ratos diabéticos.
Keywords in Portuguese
cicatrização
diabetes experimental
espectrofotometria
Abstract in Portuguese
O trabalho tem por objetivo desenvolver um protocolo experimental de avaliação da cicatrização de lesões cutâneas em ratos diabéticos (aloxânicos) através da análise do espectro de cor das fibras colágenas e de macrófagos. Considerando que em indivíduos normais, os mecanismos que regulam a cicatrização são bem conhecidos, determinou-se neste estudo verificar os mecanismos que regulam a cicatrização de animais diabéticos. Utilizamos inicialmente 60 ratos Wistar, machos. O diabetes foi induzido por injeção intravenosa (veia dorsal do pênis de Aloxana (2, 4, 5, 6 - Tetraoxypyrimidina; 5 - 6 Dioxyuracila) SIGMA, 0,1ml de solução a cada 100 g. de peso corporal. Para se chegar ao número de 12 ratos diabéticos, foram utilizados 48 ratos, destes 39,58% não desenvolveram a hiperglicemia nos néveis fixados (acima de 250mg/l ), 6,25% morreram por hipoglicemia nas primeiras 24 horas; nas 48 horas seguintes 20,83% % morreram por hiperglicemia, 4,17% morreram durante procedimento anestésico e 4,17% morreram ao longo do experimento. Foi realizada uma lesão no dorso dos animais que foram divididos em 2 grupos com 12 animais cada e receberam a denominação de Grupo 1 Diabético Grupo 2 Não Diabético. As 24 amostras dos tecidos colhidas no 3º, 7º e 14º após a lesão, foram incluídas em parafina e cortadas transversalmente e passaram por dois processos distintos, ou seja, 24 amostras por coloração com Tricrômico de Masson e 24 amostras pela técnica imunohistoquímica - anticorpo monoclonal HAM 56. Utilizou-se o software IMAGELAB para a análise morfométrica do percentual de fibras colágenas por densidade de cor e número de macrófagos evidenciados pela imunohistoquímica. Os dados obtidos em relação ao percentual de fibras colágenas e macrófagos foram tratados estatisticamente por testes paramêtricos de análise de variância, segundo dois critérios, e confirmados pelo Teste "Tukey" onde se fixou em 5% o nível de rejeição da hipótese de nulidade. Fixando-se: para ambos p< 0,5. Concluímos que o protocolo experimental para avaliação da cicatrização de lesões em ratos diabéticos se mostrou eficaz, demonstrando que a cicatrização no grupo de animais diabéticos apresentou-se retardada durante todas as etapas do processo de reparo e que a porcentagem de fibras colágenas e macrófagos pode ser facilmente analisada pelo espectro de cor.
Keywords in English
diabetes
experimental
healing
spectrophotometry
Abstract in English
This study aims to develop an experimental evaluation protocol of the healing of skin wounds in diabetic rats (aloxanic) through the analysis of color spectrum of collagenous fibres and macrophage. Considering that the mechanisms regulating scarring in normal individuals are well-known it was determined to verify the mechanisms regulating the scarring in diabetic animals. 60 Wistar male rats were used. Diabetes was induced by means of intravenous injection (dorsal penis vein of Aloxana (2, 4, 5, 6 - Tetraoxypyrimidine; 5 - 6 Dioxyuracila) SIGMA, 0,1 ml of solution per 100g of body weight. In order to reach 12 diabetic rats, 48 rats were used. 39.58% of them did not develop hypoglycemia in the established levels (above 250mg/l), 6.25% died from hypoglycemia within 24 hours; in the following 48 hours, 20.83% died from hypoglicemia, 4,17% died during the anesthetic procedures and 4,17% died along the experiment. In order to follow the study a lesion was performed on the back of the animals which were divided into two groups with 12 animals each and received the following denomination: Group I Diabetic and Group II Non-Diabetic. The 24 tissue samples collected in the 3rd , 7th and 14th days after the lesion were included in paraffine and transversally cut and suffered two different processes, that means, 24 samples received coloration with Masson Tricomic and 24 samples suffered the imunohistochemical technic - HAM 56 monoclonic antibody. The software IMAGELAB was used in the morphometric analysis of the percentage of collagen by density of colour and number of macrophage highlighted by immunehistochemistry The obtained data concerning the percentage of collagenous fibres and the number of macrophage were statistically treated by ANOVA following two criteria and confirmed by Tukey Test where the rejection level of nullity possibility was established to be 5% (alpha<=0,05). It was obtained p< 0,5 for both. We came to the conclusion that the experimental protocol for the evaluation of healing of wounds in diabetic rats has proved efficient, showing that the healing in the group of diabetic animals is slowed down during all the stages of the repairing process; the percentage of collagenous fibres and macrophage can be easily analysed by the colour spectrum.
 
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Publishing Date
2003-02-21
 
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