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Efeito citotóxico de duas resinas utilizadas em prótese removível : ProBase® Hot VS ProBase® Cold : um estudo in vitro
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Authors
Abstract(s)
Introdução: As resinas de polimetilmetacrilato são materiais essenciais no campo da Reabilitação Oral. No entanto, o seu uso tem sido posto em causa, uma vez que podem originar reações adversas nos tecidos adjacentes. Trabalhos anteriores mostraram que as resinas autopolimerizáveis libertam maiores quantidades de monómeros residuais, quando comparadas com as termopolimerizáveis. Assim, torna-se importante perceber se os métodos de polimerização têm influência na citotoxicidade desses materiais. Objetivos: Avaliar, in vitro, o nível de citotoxicidade das resinas ProBase® Hot e ProBase® Cold em fibroblastos 3T3, com a hipótese de que existem diferenças entre as resinas acrílicas ProBase®, no que diz respeito à sua citotoxicidade. Materiais e Métodos: Os espécimes foram preparados de acordo com as instruções do fabricante. Em seguida, 4 discos (5mm x 2mm) foram incubados por 30 minutos, 24 horas, 7 dias, 14 dias e 1 mês em 7mL de meio de cultura. Diluições em série dos extratos de resina previamente incubados foram colocados em contacto com 4x103 e 2x104 fibroblastos de murganho 3T3, plaqueados no dia anterior numa placa de 96 poços. Após 24 horas de exposição, a viabilidade celular foi avaliada através dos ensaios do Cristal Violeta e do MTT. A absorvância foi lida a 595nm no leitor de microplacas Bio-Rad 680. Foi utilizado o ANOVA Factorial (SPSS, Chicago, IL) para análise estatística dos resultados, aceitando-se uma significância de 5%. Resultados: Todos os fatores testados (resinas, diluições e tempo) produziram diferenças significativas na viabilidade celular. Houve diferenças estatisticamente significativas entre ambas as resinas, embora pouco significativas em termos experimentais e clínicos. A viabilidade celular nunca foi inferior a 70%, sendo este o limite estabelecido na norma ISO 10993-5:2009 para um material ser considerado citotóxico. Conclusão: Nas condições testadas, as resinas ProBase® não são citotóxicas. Existem algumas diferenças de citotoxicidade, sugerindo que o método de polimerização tem influência.
Introduction: Polymethylmethacrylate resins are essential materials in the Oral Rehabilitation field. However, their use is being called into question once they can cause adverse reactions in the adjacent tissues. Previous works have shown that autopolymerized resins release higher amounts of residual monomers than heat-polymerized resins. Having that in consideration, it is important to know if polymerization methods have influence in the cytotoxicity of these materials. Objectives: In this in vitro study, our aim is to evaluate the cytotoxicity level of ProBase® Hot and ProBase® Cold in fibroblastic cell line using cell viability assays and our hypothesis is that there are differences between ProBase® acrylic resins, regarding their cytotoxicity. Materials and Methods: Specimens of each resin were prepared according to manufacturer’s instructions. Then, 4 disks (5mm x 2mm) were immersed in 7mL of culture medium and incubated for 30 minutes, 24 hours, 7 days, 14 days and 1 month. Serial dilutions of the previously incubated resin extracts were placed in contact with 4x103 and 2x104 of 3T3 mouse fibroblasts plated in a 96 well plate the day before. After 24 hours of exposition, cell viability was evaluated through the Crystal Violet and the MTT assays. Absorvance was measured at 595nm in a microplate reader (Bio-Rad 680). The results were evaluated by Factorial ANOVA (SPSS, Chicago, IL) and the level of significance was chosen as p-value < 0.05. Results: All tested factors (resins, dilutions and time) have shown to produce significant differences in cell viability. There were statistically significant differences between both resins but not significant in experimental and clinical terms. Cell viability was never below 70%, which is the threshold stated in ISO 10993-5:2009 to be considered cytotoxic. Conclusion: Our results show that ProBase® resins are not cytotoxic in our cell model and that there were some differences in cytotoxicity, suggesting that the polymerization method has influence.
Introduction: Polymethylmethacrylate resins are essential materials in the Oral Rehabilitation field. However, their use is being called into question once they can cause adverse reactions in the adjacent tissues. Previous works have shown that autopolymerized resins release higher amounts of residual monomers than heat-polymerized resins. Having that in consideration, it is important to know if polymerization methods have influence in the cytotoxicity of these materials. Objectives: In this in vitro study, our aim is to evaluate the cytotoxicity level of ProBase® Hot and ProBase® Cold in fibroblastic cell line using cell viability assays and our hypothesis is that there are differences between ProBase® acrylic resins, regarding their cytotoxicity. Materials and Methods: Specimens of each resin were prepared according to manufacturer’s instructions. Then, 4 disks (5mm x 2mm) were immersed in 7mL of culture medium and incubated for 30 minutes, 24 hours, 7 days, 14 days and 1 month. Serial dilutions of the previously incubated resin extracts were placed in contact with 4x103 and 2x104 of 3T3 mouse fibroblasts plated in a 96 well plate the day before. After 24 hours of exposition, cell viability was evaluated through the Crystal Violet and the MTT assays. Absorvance was measured at 595nm in a microplate reader (Bio-Rad 680). The results were evaluated by Factorial ANOVA (SPSS, Chicago, IL) and the level of significance was chosen as p-value < 0.05. Results: All tested factors (resins, dilutions and time) have shown to produce significant differences in cell viability. There were statistically significant differences between both resins but not significant in experimental and clinical terms. Cell viability was never below 70%, which is the threshold stated in ISO 10993-5:2009 to be considered cytotoxic. Conclusion: Our results show that ProBase® resins are not cytotoxic in our cell model and that there were some differences in cytotoxicity, suggesting that the polymerization method has influence.
Description
Dissertação para obtenção do grau de Mestre no Instituto Universitário Egas Moniz
Keywords
Resinas acrílicas Citotoxicidade Monómeros residuais Prótese removível