Estudo citoquimico, imunocitoquimico e de analise de imagem de celulas fibroblasticas em proliferação e apoptose na ausencia e presença de colageno tipo I hiperpolimerizado

AUTOR(ES)
DATA DE PUBLICAÇÃO

1998

RESUMO

matrix which were disorganized and complexed to the DNA of apoptotic nuclei. The ordered arrangement of glycoproteins on the DNA was confirmed by TB-CEC (Toluidine blue-critical electrolyte concentration) which diferentiatesDNA ftom RNA after polarization mycroscopy analyses. V79 fibroblast cells submitted to the Feulgen reaction and image analysis of confluent cultures showed three different nuclear phenotypes classified as normal, suspected of apoptosis and apoptotic. This classification were done using morphological analysis and consideration of parameters such as DNA content (IOD), chromatin condensation in terms of optical density (OD) and nuclear area. The results showed that chromatin condensation increased when the three phenotypes were compared. A decrease of Feulgen-DNA values was observed in apoptotic cells and in cells suspected of apoptosis, revealing loss of DNA. In conclusion, this pointed out the importance of this method to characterized early phases of an apoptotic process and the possibility of inequivocal apoptosis diagnosis using this methodology when associated with others, such as cytochemical, immunocytochemical or biochemicalmethods. In the other aprroach, V79 fibroblastic cells were cultured on hyperpolymerized collagen type I substrates. The substrate showed special characteristics of self-assemleyand is an ordered structure that mimics the in vivo tendon. The long-term cell culture on this substrate was characterized by proliferation, differentiation and cell death by apoptosis analysis when compared to cell growth on coverslips. The Feulgen reaction allied to image analysis demonstrated that there is an increase in DNA content and in chromatin condensation when the cells are grow on the collagen substrate. The proliferation of V79 cells on collagen was less than observed for cells grown on a coverslips, which reflects the latency of this substrate. After the apoptosis specific TUNEL method, cell death was found to occur after longer culture periods than for cells grown on coverslips. The collagen substrate alters fibroblast proliferation and morphological aspects, characteristics that suggest cellular differentiation and the delay of apoptosis but not prevent the prevention of the apoptotic processo

ASSUNTO(S)

apoptose fibroblasto colageno

ACESSO AO ARTIGO

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