Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/290371
Type: TESE
Degree Level: Doutorado
Title: Influencia da matriz proteica do esmalte associada ao fator de crescimento insulina-simile sobre fibroblastos do ligamento periodontal humano
Author: Palioto, Daniela Bazan
Advisor: Lima, Antonio Fernando Martorelli de, 1954-2003
Abstract: Resumo: O conhecimento das diferenças morfológicas e proliferativas de fibroblastos do ligamento periodontal (FLP) e fibroblastos gengivais (FG) são fundamentais para o melhor entendimento do papel destas células nos eventos de homeostasia, doença e regeneração periodontal. O primeiro objetivo desse estudo foi comparar as características morfológicas, o potencial proliferativo e a síntese protéica de FLP e de FG. Os fibroblastos foram cultivados pela técnica do explante a partir de fragmentos gengival e do ligamento periodontal de um mesmo paciente. As células foram isoladas e plaqueadas para análise em microscopia de contraste de fase e microscopia óptica. O índice de proliferação celular foi determinado por contagem automática de células nos dias 1, 4, 7, 15 e 21 e pelo ensaio de incorporação de bromodioxiuridina (BrdU). A síntese de proteína total foi verificada por eletroforese em gel de poliacrilamida e zimografia. Os resultados mostraram que FLP são maiores e mais alongados que os FG em condições de subconfluência e confluência celular. Os FLP proliferam mais rapidamente que os FG nos períodos de 1, 4 e 7 dias (P<0,05). Nos períodos subsequentes de 15 e 21 dias, não houve diferença estatística significativa entre o número de células do dois tipos celulares. O índice de incorporação de BrdU demonstrou potencial proliferativo de 65,1 .:t 11,0 % para FLP e 41,2.:t 17,1% para FG (P<0,05). A síntese de proteína total verificada em nosso experimento no período de 24 horas mostrou resultados similares para FLP e FG. Nossos resultados demonstraram que FLP e FG são diferentes na morfologia e na capacidade proliferativa, porém são semelhantes na síntese protéica. O segundo objetivo deste estudo foi avaliar os efeitos da matriz protéica do esmalte (EMD), do fator de crescimento insulina-símile (IGF-I) e da associação dos dois fatores sobre o potencial proliferativo e a capacidade de adesão e migração celular de fibroblastos do ligamento periodontal humano. Esse estudo foi conduzido utilizando a análise de proliferação por contagem automática de células (Coulter Counter_

Abstract: The knowledge of morphological and proliferative differences between Gingival Fibroblasts (GF) and Periodontal Ligament Fibroblast (PDL) is essential to understand the role of each cell on the homeostasis, disease process and regeneration Df the periodontium. The first objective of this study was to compare the PDL and GF morphology, proliferation rate and protein synthesis. PDL and GF were explanted from tissues Df the same patients. To characterize and compare the cells morphology, PDL and GF were plated for optical and phase contrast microscopy. The proliferation rates were determined by automated count at days 1, 4, 7, 15 and 21, and by Bromodeoxyuridine Labelling Index (BrdU). The total protein content was analyzed by means of electrophoresis in 10% polyacrylamide gel and zimography containing gelatin as substrate. PDL were greater and more elongatOO than GF in subconfluence and confluence conditions. The proliferative rate Df PDL was higher than GF at 1, 4, and 7 days (P < 0.05). At 15 and 21 days there was no statistically significant difference between cells number. The BrdU Index demonstrated a proliferative potential Df 65.1 ! 11.0 % for PDL and 41.2 :t 17.1 % for GF (P < 0.05).The synthesis of protein in a period of 24 hours was similar for both PDL and GF. Our results demonstrated that PDL and GF are different in morphology and in proliferative capacity, however, they do not differ in protein synthesis. The second objective of the present study was to evaluate the effects of the enamel matrix protein (EMD) the effects of Insulin growth factor-I (IGF-I) and the association of the these two factors on periodontal ligament fibroblasts. The study was based on the proliferation rate, and cellular attachment and migration. The proliferation rate was determined by counting the cells afier 1, 3, 7 and 10 days with the help of a automated counter (Coulter Counter). The cellular attachment was analyzed by means of colorimetric assays. Migration assays was performed in Boyden chambers for a period of 4h and analyzed by means of colorimetric assays. EMD has significantly raised the proliferation rate from a minimum concentration of 50 uglml up to maximum concentration of 200 uglml in a 24h period. The proliferation rate continuing to be raised in a period of 3,7 and 10 days (P < 0.05). IGF-I has not altered the proliferation rate at any concentration. The association of EMD + IGF-I raised the proliferation rate independently of the concentration. However, the association has not resulted on further significant effect when compared to the use of EMD alone. The cellular attachment and migration was not altered by EMD, IGF-I or EMD + IGF-I. Our results showed that EMD increased the proliferation rate of PDL but did not improve cellular attachment and migration. IGF-I did not raise neither the proliferation rate no r the cellular attachment and migration and did not have any additional effect on the EMD alone
Subject: Células
Matriz extracelular
Cultura celular
Language: Português
Editor: [s.n.]
Date Issue: 2001
Appears in Collections:FOP - Tese e Dissertação

Files in This Item:
File SizeFormat 
Palioto_DanielaBazan_D.pdf2.45 MBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.