Análise da citotoxicidade do agregado de trióxido mineral (MTA-Branco) em cultura de odontoblastos. Influência dos tempos de presa e do armazenamento do material em meio líquido
Analysis of the cytotoxicity of a mineral trioxide aggregate (White-MTA) on cultured odontoblasts. Influence of the setting time and storage of the material in wet environment
Frederico, P.G.; Lanza, C.R.M.; Aranha, A.M.F.; Hebling, J.; Costa, C.A.S.
Rev. odontol. UNESP, vol.35, n4, p.319-326, 2006
Resumo
O agregado de trióxido mineral - MTA-Branco (Angelus - Ind. Com. Ltda, Br.) é um material nacional para uso na clínica endodôntica que apresenta custo inferior a outros cimentos MTA disponíveis no mercado. Como pouco se sabe de seus efeitos sobre células pulpares, o objetivo da presente pesquisa foi avaliar a citotoxicidade dessa formulação de MTA em cultura de células odontoblastóides MDPC-23. Corpos-de-prova foram preparados e imersos em meio de cultura (DMEM) puro após 1 hora (presa incompleta) ou 4 horas (presa completa) de sua manipulação, onde permaneceram pelos períodos de 24 horas ou 7 dias, determinando os grupos experimentais seguintes: G1: 1 hora de tempo de presa/24 horas de imersão; G2: 4 horas de tempo de presa/24 horas de imersão; G3: 1 hora de tempo de presa/7 dias de imersão; e G4: 4 horas de tempo de presa/7 dias de imersão. Células MDPC-23 (30.000 células/cm2) foram cultivadas em placas de 24 compartimentos, e, após 72 horas de incubação, o meio DMEM foi aspirado e substituído pelos extratos obtidos do MTA-Branco. Nos grupos controle (G5 e G6), DMEM puro foi utilizado. O pH dos extratos e o do DMEM puro foram determinados, mostrando baixa variação entre os grupos (de 8,4 até 8,9). O efeito citotóxico foi avaliado por meio da análise colorimétrica do Metiltetrazolium (teste do MTT), e os resultados obtidos não mostraram diferenças estatisticamente significantes entre os grupos experimentais e entre estes e os controles, já que a redução do metabolismo para os grupos G1, G2, G3 e G4 foi, respectivamente, de 0,82; 9,6; 4,4 e 0%. A análise da morfologia celular em microscopia eletrônica de varredura (MEV) mostrou não haver diferenças entre os grupos experimentais e controles. Baseado nos resultados obtidos foi possível concluir que o tempo de presa e o de armazenagem do cimento MTA-Branco em líquido não interferem negativamente no metabolismo celular, caracterizando o insignificante efeito citotóxico do material.
Palavras-chave
Materiais dentários, agregado de trióxido mineral, cimentos dentários, citotoxicidade.
Abstract
The mineral trioxide aggregate - white-MTA (W-MTA, Angelus Ind. Com. Ltda, Br.) is a hard-setting cement widely used in endodontics. This dental material presents lower cost than some similar cements available in dentistry. Since the cytotoxicity of this dental material to the pulp cells is unknown, the aim of this in vitro study was to evaluate the toxic effects of the W-MTA to an immortalized odontoblast cell line (MDPC-23). Round-shaped samples were prepared and immersed in culture medium (DMEM) 1 hour (non-set material) or 4 hours (set material) after hand-mixing. The samples remained in DMEM for 24 hours or 7 days, giving rise to the following experimental groups: G1: 1 hour set period/24hours immersion; G2: 4hours set period/24 hours immersion; G3: 1 hour set period/7-day immersion; and G4: 4 hours set period/7- day immersion. After 72 hours incubation, the complete DMEM that was in contact with the cells (30,000 cells/cm2) was replaced by the extracts obtained from the experimental W-MTA. In control groups (G5 and G6) fresh DMEM was used. The cell metabolic activity was evaluated by the Methyltetrazolium (MTT) assay which determines the mitochondrial respiration of the cells. In G1, G2, G3, and G4 the cell metabolism decreased by 0.82; 9.6; 4.4 and 0%, respectively. The statistical analysis showed no significant difference among the experimental and control groups. The cell morphology evaluated by scanning electron microscope (SEM) determined no difference among the experimental and control groups. Based on the experimental conditions, it is possible to conclude that the W-MTA presents non significant cytopathic effects to the MDPC-23 pulp cells. The immersion of set or non-set W-MTA round-shaped samples in wet environment does not influence the cytotoxicity of this hard-setting cement.
Keywords
Dental materials, mineral trioxide aggregate, dental cements, cytotoxicity
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