Influencia da matriz proteica do esmalte associada ao fator de crescimento insulina-simile sobre fibroblastos do ligamento periodontal humano

AUTOR(ES)
DATA DE PUBLICAÇÃO

2001

RESUMO

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

ASSUNTO(S)

matriz extracelular cultura celular celula

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