REGIANE MARTINS
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Artigo IPEN-doc 27057 Evaluation of cytotoxicity through MTS test of 2-ACBs (2-dDCB AND tDCB) after exposure to hepatic cells2019 - BARBEZAN, A.B.; CARVALHO, L.R.; VIEIRA, D.P.; MACHADO-SANTELLI, G.M.; MARTINS, R.; VILLAVICENCIO, A.L.C.H.Food irradiation has been approved in more than 60 countries for many applications in a wide variety of foods. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products formed when foods containing fatty acids are irradiated. Despite the importance of food irradiation, the toxicological potential of 2-ACBs in irradiated food is still no fully understood. In this study we investigated the cytotoxic effects of irradiated palmitic and stearic fatty acids byproducts, 2-dDCB and 2-tDCB, in hepatic cell lines (HepG2, BRL3A and HTC). The cytotoxic effects of 2-dDCB and 2-tDCB were evaluated at 100, 300 and 500 μM for 24 and 48 hours and the cell viability was measured using the MTS assay. While no toxicity was observed for 2-tDCB in all cells for all tested conditions, 2-dDCB was found to be toxic to BRL3A cells (at 100 μM after 48 hours) and HTC cells (at 24 hours in all tested concentrations). HepG2 cells on the other hand, were found to be resistant to 2-dDCB-induced toxicity. Overall our data shows that the byproduct 2-tDCB is not toxic for hepatic cells while 2-dDCB can be used has a potential marker for food irradiation-induced toxicity.Artigo IPEN-doc 27906 In vivo genotoxicity of 2-alkylcyclobutanones in liver cells from rats fed with irradiated cocoa butter using flow cytometry2021 - MARTINS, R.; VIEIRA, D.P.; CARVALHO, L.R.; BARBEZAN, A.B.; VILLAVICENCIO, A.L.C.H.Food irradiation proves to be an effective technique of eliminating some pathogens from food and this has gained significant attention to its potential for food safety. Since 1990, studies on the toxicological safety of 2-Alkylcyclobutanones have been conducted extensively. 2-Alkylcyclobutanones are unique radiolytic products generated by the radiation-induced breakage of triglycerides in food, 2-dodecylcyclobutanone (2-dDCB) and 2-tetradecylcyclobutanone (2-tDCB) are the predominant compounds detected . Despite studies showing non-genotoxicity of 2-ACBs (2-Alkylcyclobutanones), the results are conflicting and therefore we continue the studies in order to confirm the compounds are safety for human health. In vivo micronucleus test were performed to verify the 2-ACBs genotoxic effects in hepatic cells using flow cytometry. We used cocoa butter irradiated with 20 kGy. A group with animals was treated with daily intake of irradiated cocoa butter, synthesized 2-dDCB and 2-tDCB for one month. Hepatic cells were selected for genotoxicity analysis due to the liver importance in the compounds metabolization. The improvement of the analytical techniques is important for the research future since the irradiation process is already consolidated. The results confirmed the safety of the food irradiation process, as they did not indicate the genotoxic potential of the samples.Resumo IPEN-doc 26930 Avaliação do potencial citotóxico da 2-tetradecilciclobutana em células hepáticas linhagem HepG22016 - BARBEZAN, A.B.; SALES, B.R.; MARTINS, R.; BUENO, J.B.; SANTELLI, G.M.M.; VILLAVICENCIO, A.L.C.H.Introdução: A irradiação de alimentos é um método eficaz e seguro para a preservação e armazenamento em longo prazo, é aprovado para utilização em mais de 60 países para diversas aplicações em uma ampla variedade de produtos alimentares (Agric. Food Chem. 51; 927, 2003. Food Chem. 201; 52-58, 2016). Este processo é realizado através da utilização de feixes de elétrons acelerados, raios-X ou radiação gama (60Co ou 137Cs). As 2-Alcilciclobutanonas (2-ACBs) são os únicos produtos radiolíticos conhecidos gerados a partir de alimentos que possuem ácidos graxos (Triglicérides) e são submetidos à irradiação (J. Food Protc. 67; 142, 2004. T. Food Scie.Tech. 44; 66-78, 2015). O ácido analisado neste estudo é o esteárico que quando irradiado forma 2-Tetradecilciclobutanona (2-tDCB). Desde a década de 1990 estudos toxicológicos de segurança das 2-ACBs tem sido conduzido extensivamente através de compostos sintéticos. Testes de mutagenicidade das 2-ACBs realizados indicam claramente que nenhuma evidência foi observada, enquanto estudos de viabilidade apresentaram citotoxicidade notada através da morte celular (Food Scie. Tech. 44; 66-78, 2015). Parte das 2-ACBs ingeridas é excretada através das fezes e parte ficam depositadas em tecidos adiposos. Estudos realizados até o momento foram somente em células de cólon. A linhagem escolhida para este trabalho é derivada de células hepáticas uma vez que o acumulo de gordura neste órgão é bastante comum. Objetivo: Avaliar possíveis danos citotóxicos, através do teste de viabilidade celular MTT observando a influência de diversas concentrações da 2-tDCB em diferentes tempos de incubação em células hepáticas da linhagem HepG2. Métodos: O composto 2-tDCB foi solubilizado em etanol a 2%. A linhagem celular escolhida é derivada de hepatocarcinoma humano (HepG2) e foi cultivada em meio de cultura suplementado com 10% de soro fetal bovino. As células foram plaqueadas na densidade de 5x103 cél/poço em uma placa de 96 poços. O efeito citotóxico da 2-tDCB foi avaliado nas concentrações de 100, 300 e 500μM, durante 24 e 48 horas. Os testes foram realizados de acordo com instruções do kit CellTiter96 Aqueus Non-radioactive Cell Proliferation Assay, em triplicatas (biológica e experimental) e os resultados foram analisados pelo programa Prisma GraphPad. Resultados: A linhagem tratada com 2-tDCB em 24 e 48h não apresentou citotoxicidade em nenhuma das concentrações avaliadas. Conclusão: Não houve inviabilidade causada pelo composto 2-tDCB na linhagem de células hepática estudadas, nenhum dano foi observado em nenhuma das variações pesquisadas. Estudos mais aprofundados são necessários para identificar os mecanismos moleculares pela qual o composto em questão atua.Artigo IPEN-doc 26253 In vivo genotoxicity of 2-Alkylcyclobutanones in liver cells from rats fed with irradiated cocoa butter using flow citometry2019 - MARTINS, REGIANE; VIEIRA, DANIEL P.; CARVALHO, LUMA R. de; BARBEZAN, ANGELICA B.; VILLAVICENCIO, ANNA L.C.H.Food irradiation proves to be an effective technique of eliminating some pathogens from food and this has gained significant attention to its potential for food safety. Since 1990, studies on the toxicological safety of 2-Alkylcyclobutanones have been conducted extensively. 2- Alkylcyclobutanones are unique radiolytic products generated by the radiation-induced breakage of triglycerides in food, are exclusively found in irradiated lipid containing foods. 2-dodecylcyclobutanone (2-dDCB) and 2-tetradecylcyclobutanone (2-tDCB) are the predominant compounds detected in irradiated food. Despite studies showing non- genotoxicity of 2-ACBs (2-Alkylcyclobutanones), the results are conflicting and therefore we continue the studies in order to confirm the compounds safety for human health. In vivo micronucleus test were performed to verify the 2-ACBs genotoxic effects in hepatic cells using flow citometry. We used cocoa butter irradiated with 20 kGy at IPEN GAMACELL. A group with animals (IPEN Ethical Animal Experimentation Committee, process number 148/14) was treated with daily intake of irradiated cocoa butter, synthesized 2- Dodecylcyclobutanone and 2-Tetradecylcyclobutanone for two months. Hepatic cells were selected for genotoxicity analysis due to the liver importance in the compounds metabolization. Analyzes were made by micronucleus test with specific cells extracted from hepatic tissue using flow cytometry, which is an alternative to conventional techniques, allowing faster analysis and reduction in the animals number that is a subject much approached in research today. The improvement of the analytical techniques is important for the research future since the irradiation process is already consolidated. The results confirmed the safety of the food irradiation process, as they did not indicate the genotoxic potential of the samples.Resumo IPEN-doc 23504 Avaliação do potencial citotóxico da 2-tetradecilciclobutana em células hepáticas linhagem HepG2– estudos in vitro2016 - BARBEZAN, A.B.; SALES, B.R.; MARTINS, R.; BUENO, J.B.; SANTELLI, G.M.M.; VILLAVICENCIO, A.L.C.H.