Efeito da nicotina na viabilidade e morfologia de fibroblastos: estudo in vitro
Carregando...
Data
2002-09-01
Autores
Martinez, Aurora Esmeralda Traverso [UNESP]
Silvério, Karina Gonzales [UNESP]
Rossa Júnior, Carlos [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Sociedade Brasileira de Pesquisa Odontológica (SBPqO)
Resumo
O objetivo do estudo foi avaliar in vitro o efeito da nicotina sobre a viabilidade e a morfologia celular utilizando-se uma linhagem contínua de fibroblastos. Para tal, foram formados dois grupos experimentais segundo a dose (0 - controle, 10 mig, 100 mig, 0,5 mg, 1 mg) e o tempo de condicionamento (1 e 24 horas). Cada um dos 12 orifícios de uma placa para cultura celular recebeu 2 ml de meio de Eagle, e 1 ml de suspensão de meio de cultura contendo aproximadamente 1 × 10(5) células/ml. Foi, então, acrescentada a solução de nicotina nas diferentes concentrações. Após o condicionamento com a droga, nos dois períodos testados, as células foram coradas com azul de trypan 0,4%, e observadas em microscópio invertido por um examinador cego para os grupos experimentais, que avaliou a viabilidade e a morfologia segundo o índice de Gamal. Os experimentos foram repetidos 5 vezes. Quanto à morfologia, os resultados obtidos demonstraram, no grupo condicionado por 1 h, que os controles apresentaram diferenças estatisticamente significantes em relação apenas à maior dose de nicotina; no entanto, foram encontradas diferenças estatisticamente significantes entre o controle e todas as concentrações após 24 horas de condicionamento. Na viabilidade celular, um maior número de células não viáveis foi observado nas diferentes concentrações de nicotina em comparação aos controles tanto após 1 quanto 24 horas de condicionamento (p < 0,05). em ambos períodos existiu uma tendência significativa de aumento do número de células não viáveis com o aumento da dose de nicotina (p = 0,0053; p = 0,00001 após 1 e 24 h respectivamente). Portanto, conclui-se que a nicotina pode alterar, in vitro, a viabilidade e a morfologia de fibroblastos de forma proporcional à dose e ao tempo de exposição.
The aim of this study was to evaluate, in vitro, the effect of nicotine on the viability and morphology of fibroblasts from a continuous lineage. Two experimental groups were prepared, with different drug dosages (0 - control, 10 mug, 100 mug, 0.5 mg, 1 mg) and conditioning time (1 and 24 hours). Twelve-well microplates were utilized. Each well received 2 ml of fresh culture medium and 1 ml of a solution containing 1 × 10(5) cells/ml. Nicotine was then added to the wells, at the tested concentrations. After the incubation period, cell viability was assessed by means of 0.4% trypan blue staining. Cell viability and morphology were assessed in an inverted microscope, by a single examiner, who was blind as to the experimental groups. The experiment was repeated 5 times. Regarding morphology, in the 1-hour conditioned group there was statistically significant difference between the control group and the group with the greatest dose of nicotine. These differences were also observed between the control group and all nicotine groups after 24 hours. The results of the Kruskal-Wallis test revealed that more unviable cells were found in the groups exposed to nicotine, in comparison with the control group, both after 1 and 24 hours of conditioning (p < 0.05). Moreover, with increasing doses of nicotine there was a directly proportional increase in the number of unviable cells, both after 1 and 24 hours of exposure (p = 0.0053 and p = 0.00001, respectively). The conclusion of this study is that nicotine can alter, in vitro, the viability and morphology of fibroblasts in a manner proportional to the dose and time of exposure.
The aim of this study was to evaluate, in vitro, the effect of nicotine on the viability and morphology of fibroblasts from a continuous lineage. Two experimental groups were prepared, with different drug dosages (0 - control, 10 mug, 100 mug, 0.5 mg, 1 mg) and conditioning time (1 and 24 hours). Twelve-well microplates were utilized. Each well received 2 ml of fresh culture medium and 1 ml of a solution containing 1 × 10(5) cells/ml. Nicotine was then added to the wells, at the tested concentrations. After the incubation period, cell viability was assessed by means of 0.4% trypan blue staining. Cell viability and morphology were assessed in an inverted microscope, by a single examiner, who was blind as to the experimental groups. The experiment was repeated 5 times. Regarding morphology, in the 1-hour conditioned group there was statistically significant difference between the control group and the group with the greatest dose of nicotine. These differences were also observed between the control group and all nicotine groups after 24 hours. The results of the Kruskal-Wallis test revealed that more unviable cells were found in the groups exposed to nicotine, in comparison with the control group, both after 1 and 24 hours of conditioning (p < 0.05). Moreover, with increasing doses of nicotine there was a directly proportional increase in the number of unviable cells, both after 1 and 24 hours of exposure (p = 0.0053 and p = 0.00001, respectively). The conclusion of this study is that nicotine can alter, in vitro, the viability and morphology of fibroblasts in a manner proportional to the dose and time of exposure.
Descrição
Palavras-chave
Nicotina, efeitos adversos, Fibroblastos, Tabaco, Nicotine, adverse effects, Fibroblasts, Tobacco
Como citar
Pesquisa Odontológica Brasileira, v. 16, n. 3, p. 234-238, 2002.