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Doctoral Thesis
DOI
https://doi.org/10.11606/T.5.2011.tde-23032011-165803
Document
Author
Full name
Elisabeth Mie Hosaka
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Mathor, Monica Beatriz (President)
Fernandes, Tulio Diniz
Moreira Filho, Carlos Alberto
Schiozer, Wandir Antonio
Termini, Lara
Title in Portuguese
Comparação de modificações nos comportamentos celular e gênico de fibroblastos derivados de úlcera de membro inferior em indivíduos diabéticos e de pele normal
Keywords in Portuguese
Diabetes mellitus
Expressão gênica
Fibroblastos
Pé diabético
Abstract in Portuguese
Úlcera de membros inferiores é uma das complicações do diabetes melito que acomete aproximadamente 15% dos portadores da doença e resultam em elevada taxa de mortalidade. São feridas de difícil resolução, em geral, refratárias aos diversos tipos de tratamentos. Um dos principais aspectos que contribuem para sua cronicidade é a modificação no comportamento de fibroblastos. O objetivo do estudo foi comparar in vitro modificações no comportamento de fibroblastos derivados de ferida de membro inferior de indivíduos diabéticos com fibroblastos derivados de pele normal, quanto a proliferação celular e a capacidade de contração de modelo tridimensional de matriz de colágeno povoado por células, além de investigar o perfil de expressão gênica diferencial dessas células. Para tanto, foram cultivados fibroblastos provenientes de tecido de granulação de feridas de membro inferior de pacientes com diabetes do tipo 2 (FFD) e de pele normal (FC). Foram verificadas anormalidades morfológicas no grupo FFD compatíveis com senescência celular; menor contração da matriz de colágeno povoado por fibroblastos do grupo FFD (redução de 23% da área original) comparado ao grupo FC (redução de 30% da área original p<0,001). Pela técnica de microarray foram identificados 143 genes diferencialmente expressos, em sua maioria hipoexpressos, no grupo FFD, dentre os quais, destacam-se aqueles relacionados a processos de senescência e apoptose celular, bem como déficit na síntese e contração de fibras colágenas
Title in English
Comparision of changes in cell behavior and genic of fibroblast derived from leg ulcers in diabetic and normal skin
Keywords in English
Diabetes mellitus
Diabetic foot
Fibroblast
Gene expression
Abstract in English
Lower limbs ulcer is one of the complications of diabetes mellitus that affects approximately 15% of the patients and results in high mortality. Wounds that is difficult to heal, generally refractory to many treatments. One of the main aspects that contribute to disease chronicity is the change in the fibroblast behavior. The aim of this study was to compare changes in the in vitro fibroblasts behavior from diabetic leg ulcer with fibroblasts derived from normal skin as to cell proliferation and contraction capacity of a threedimensional fibroblasts populated collagen lattice, as well as to investigate the pattern of differential gene expression in these cells. For this purpose, fibroblasts were cultured from granulation tissue of lower limb ulcer in patients with type 2 diabetes (FFD) and from normal skin (FC). Morphological abnormalities compatible with cellular senescence were observed in the FFD. Reduced matrix contraction in fibroblasts populated group (FFD reduction of 23% from the original area) compared to FC (30% reduction from the original area) - (p <0.001). Using DNA microarray 143 differentially expressed genes were identified, most of them were underexpressed in FFD group, among which are those related to senescence and apoptosis, as well as deficits in the synthesis and contraction of collagen fibers
 
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Publishing Date
2011-03-24
 
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