Avaliação in vitro da mucograft® em fibroblastos gengivais humano
Rafaela Silva Rezende e Lima.
Dissertação
por
D64
Campinas : [s.n.], 2014.
63 f. : il.
Dissertação (Mestrado em Periodontia) - Centro de Pesquisas OdontológicasSão Leopoldo Mandic.
A Mucograft® é uma matriz reabsorvível, de origem suína, composta de colágeno do tipo I e III, produzida em duas camadas e indicada como substituto para enxertos conjuntivos. Este estudo teve como objetivo avaliar in vitro, o comportamento biológico da Mucograft® em fibroblastos gengivais humanos....
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A Mucograft® é uma matriz reabsorvível, de origem suína, composta de colágeno do tipo I e III, produzida em duas camadas e indicada como substituto para enxertos conjuntivos. Este estudo teve como objetivo avaliar in vitro, o comportamento biológico da Mucograft® em fibroblastos gengivais humanos. Foi avaliada a proliferação e viabilidade celulares, utilizando-se os métodos de exclusão vital azul de Trypan e MTT, respectivamente, após 1, 2, 3 e 7 dias. Além disso, foi avaliada a expressão de duas proteínas abundantes da matriz extracelular, o colágeno tipo III (COL 3A1) e vimentina (VIM), através de Western-blotting, após 10 e 14 dias. Os resultados demonstraram que os fibroblastos gengivais humanos, cultivados sobre as membranas reabsorvíveis Mucograft® (grupo teste, M), apresentaram proliferação e viabilidade semelhantes em comparação às células que foram cultivadas apenas no meio de cultura, pertencentes ao grupo controle (C), em todos os tempos estudados, sem apresentar diferença estatística significante entre os grupos (p>0,05). Em relação à expressão semi-quantitativa de COL 3A1 e VIM, através de Western-blotting, após 14 dias, nota-se uma diminuição da expressão de COL 3A1 quando as células foram cultivadas sobre a matriz (M), com significância estatística entre os grupos (p<0,05), sem diferenças estatísticas para VIM entre os grupos e tempos estudados. Com base nos resultados obtidos, pode-se concluir que os fibroblastos gengivais, quando cultivados sobre a matriz reabsorvível (teste), apresentaram proliferação e viabilidade celulares similares ao controle. Além disso, as análises da síntese proteica mostraram que esta matriz não apresentou função indutora na síntese de COL 3A1 e VIM, mas evidenciaram, principalmente, aos 14 dias uma diminuição da síntese de COL 3A1, indicando uma possível função essencialmente de arcabouço.
Palavras-chave: Enxerto gengival. Matriz de colágeno Mucograft®. Cultura celular. Biocompatibilidade.
The Mucograft® is a resorbable matrix of porcine origin, composed by type I and III collagen, produced in two layers and it is indicated to replace connective grafts. It can be used to increase soft tissue around teeth and implants, in guided bone regeneration, reconstruction of the alveolar ridge, closing of alveoli, to cover exposed bone tissue, as well as, root coverage. This study aimed to evaluate, in vitro, the biocompatibility of Mucograft® Collagen Matrix and differentiation induction of human gingival fibroblasts. Cell proliferation and viability were assessed by Trypan exclusion method and MTT test, respectively, after 1, 2, 3 and 7 days. In addition, type III collagen (COL 3A1) and vimentin (VIM) proteins expression was evaluated by Western blotting, after 10 and 14 days. The results demonstrated that human gingival fibroblasts in the control group (C), which were cultured in the culture medium only, showed similar results, concerning proliferation and viability, when compared to cells that were cultured on Mucograft® resorbable matrix (test group, M), at all studied periods (p> 0.05). However, after 14 days, it was observed a decrease in the expression of COL 3A1 when the cells were cultured on the matrix (M), with statistical significance (p <0.05). For VIM, it is observed that, in both studied periods, 10 and 14 days, no statistical differences (p> 0.05). Based on the results, it can be conclude that the gingival fibroblasts when cultured on resorbable matrix (test) showed proliferation and cell viability similar to control. Furthermore, analysis of the synthesis of type III and vimentin matrix proteins demonstrated that Mucograft® matrix (M) had no role on COL 3A1 and VIM synthesis induction, but showed mainly at 14 days decreased synthesis of COL 3A1, indicating a possible function essentially as a framework.
Keywords: Gingival tissue graft. Mucograft® Collagen Matrix. Cell culture. Biocompatibility. Ver menos
Palavras-chave: Enxerto gengival. Matriz de colágeno Mucograft®. Cultura celular. Biocompatibilidade.
The Mucograft® is a resorbable matrix of porcine origin, composed by type I and III collagen, produced in two layers and it is indicated to replace connective grafts. It can be used to increase soft tissue around teeth and implants, in guided bone regeneration, reconstruction of the alveolar ridge, closing of alveoli, to cover exposed bone tissue, as well as, root coverage. This study aimed to evaluate, in vitro, the biocompatibility of Mucograft® Collagen Matrix and differentiation induction of human gingival fibroblasts. Cell proliferation and viability were assessed by Trypan exclusion method and MTT test, respectively, after 1, 2, 3 and 7 days. In addition, type III collagen (COL 3A1) and vimentin (VIM) proteins expression was evaluated by Western blotting, after 10 and 14 days. The results demonstrated that human gingival fibroblasts in the control group (C), which were cultured in the culture medium only, showed similar results, concerning proliferation and viability, when compared to cells that were cultured on Mucograft® resorbable matrix (test group, M), at all studied periods (p> 0.05). However, after 14 days, it was observed a decrease in the expression of COL 3A1 when the cells were cultured on the matrix (M), with statistical significance (p <0.05). For VIM, it is observed that, in both studied periods, 10 and 14 days, no statistical differences (p> 0.05). Based on the results, it can be conclude that the gingival fibroblasts when cultured on resorbable matrix (test) showed proliferation and cell viability similar to control. Furthermore, analysis of the synthesis of type III and vimentin matrix proteins demonstrated that Mucograft® matrix (M) had no role on COL 3A1 and VIM synthesis induction, but showed mainly at 14 days decreased synthesis of COL 3A1, indicating a possible function essentially as a framework.
Keywords: Gingival tissue graft. Mucograft® Collagen Matrix. Cell culture. Biocompatibility. Ver menos
Avaliação in vitro da mucograft® em fibroblastos gengivais humano
Rafaela Silva Rezende e Lima.
Avaliação in vitro da mucograft® em fibroblastos gengivais humano
Rafaela Silva Rezende e Lima.
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