Efeitos toxicogenômicos do MTA Fillapex em fibroblastos humanos in vitro
Carregando...
Data
2020-02-18
Autores
Leme, Kamila Sauer Veiga
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
A odontologia sempre preconizou o uso de materiais biocompatíveis para não causar agressão aos tecidos sadios adjacentes. Um dos materiais mais utilizados na endodontia é o cimento endodôntico para obturação, utilizado juntamente à guta-percha para preenchimento dos canais radiculares. No entanto, compostos presentes na fórmula de alguns cimentos podem apresentar efeitos tóxicos desde o momento em que foram inseridos, até meses ou anos após o tratamento. Um dos cimentos produzidos com o objetivo de ser biologicamente compatível e com propriedades físico-químicas adequadas foi o Mineral Trióxido Agregado Fillapex (MTA Fillapex®), que foi desenvolvido a partir do cimento Mineral Trióxido Agregado (MTA), o qual possui as propriedades biológicas desejadas. Considerando os resultados divergentes sobre a citotoxicidade do MTA Fillapex®, o presente estudo teve como objetivo investigar, in vitro, os efeitos citotóxicos e toxicogenômicos desse composto em linhagem celular de fibroblastos humanos gengivais. Para tanto, foram avaliados a sobrevivência e ciclo celular, os níveis de apoptose e necrose, o potencial genotóxico (teste do cometa) e o padrão de expressão (qRT-PCR) de genes relacionados à apoptose e proliferação celular (BCL-2, BAX e TP53). Os resultados mostraram efeito citotóxico significativo do cimento nas concentrações 20% e 40%, muito embora apenas na concentração mais alta o efeito persistiu por até 5 dias após o tratamento das células (teste de sobrevivência). Por outro lado, o MTA Fillapex® apresentou efeito genotóxico em concentrações menores, de 5% e 20%, e efeito toxicogenômico, modulando a expressão dos genes BCL-2, BAX e TP53, na concentração de 20%. Além disso, foram observadas alterações na cinética do ciclo celular em todas as fases e taxas de apoptose inicial, apoptose tardia e necrose na concentração de 40% e apoptose inicial na de 20% em comparação ao grupo controle. Concluindo, o cimento MTA Fillapex® apresentou potencial citotóxico e toxicogenômico em fibroblastos humanos gengivais in vitro, indicando a necessidade da realização de outros testes, com diferentes desenhos experimentais, a fim de evitar riscos inerentes ao seu uso na odontologia.
Dentistry has always advocated the use of biocompatible materials to avoid damage to adjacent healthy tissues. One of the most commonly used endodontic material is the endodontic cement, which is used together with gutta-percha to fill root canals after pulp tissue removal. However, compounds in the formula of some cement may have toxic effects from the time they were inserted to months or years after treatment. The Mineral Trioxide Aggreate Fillapex (MTA Fillapex®), which was developed from the Mineral Trioxide Aggregate (MTA) cement, is one of the biologically compatible cements with adequate physicochemical properties. Considering the divergent results about MTA Fillapex® toxicity, the present study aimed to investigate the in vitro cytotoxicity and toxicogenomic potential of this compound in human gingival fibroblasts cell line. Cell cycle and survival, apoptosis and necrosis rates, genotoxicity (comet assay) and gene expression profiling (BCL-2, BAX, TP53) were evaluated. The results showed significant cytotoxicity of the cement at concentrations 20% and 40%, although only the effect induced at the highest concentration persisted 5 days after cell treatment (survival test). MTA Fillapex® genotoxicity was observed for treatments at concentrations of 5% and 20% and toxicogenomic effects on BCL-2, BAX and TP53 genes were detected at 20%. Changes in the cell cycle kinetics and in the rates of initial and late apoptosis and necrosis were observed at 40%. In conclusion, MTA Fillapex® presented cytotoxic and genotoxic effects, and also induced increased expression of genes related to apoptosis regulation.
Dentistry has always advocated the use of biocompatible materials to avoid damage to adjacent healthy tissues. One of the most commonly used endodontic material is the endodontic cement, which is used together with gutta-percha to fill root canals after pulp tissue removal. However, compounds in the formula of some cement may have toxic effects from the time they were inserted to months or years after treatment. The Mineral Trioxide Aggreate Fillapex (MTA Fillapex®), which was developed from the Mineral Trioxide Aggregate (MTA) cement, is one of the biologically compatible cements with adequate physicochemical properties. Considering the divergent results about MTA Fillapex® toxicity, the present study aimed to investigate the in vitro cytotoxicity and toxicogenomic potential of this compound in human gingival fibroblasts cell line. Cell cycle and survival, apoptosis and necrosis rates, genotoxicity (comet assay) and gene expression profiling (BCL-2, BAX, TP53) were evaluated. The results showed significant cytotoxicity of the cement at concentrations 20% and 40%, although only the effect induced at the highest concentration persisted 5 days after cell treatment (survival test). MTA Fillapex® genotoxicity was observed for treatments at concentrations of 5% and 20% and toxicogenomic effects on BCL-2, BAX and TP53 genes were detected at 20%. Changes in the cell cycle kinetics and in the rates of initial and late apoptosis and necrosis were observed at 40%. In conclusion, MTA Fillapex® presented cytotoxic and genotoxic effects, and also induced increased expression of genes related to apoptosis regulation.
Descrição
Palavras-chave
Apoptose, BAX, BCL-2, Ciclo Celular, Genotoxicidade