ANGELICA BUENO BARBEZAN

ANGELICA BUENO BARBEZAN

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Evaluation of cytotoxicity through MTS test of 2-ACBs (2-dDCB AND tDCB) after exposure to hepatic cells
2019 - 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.
Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2
2019 - BARBEZAN, ANGELICA B.; SANTOS, CARLA J.B.; CARVALHO, LUMA R.; VIEIRA, DANIEL P.; SANTELLI, GLAUCIA M.M.; VILLAVICENCIO, ANNA L.C.H.
Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2- Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line.
In vivo evaluation of genotoxic potential of liver cells from rats fed with irradiated diet using flow cytometry
2017 - MARTINS, R.; CARVALHO, L.R. de; VIEIRA, D.P.; BARBEZAN, A.B.; VILLAVICENCIO, A.L.C.H.
Radiation sources became widely available since 1960’s, and between its main uses are the applications in food irradiation and research of effects of ionizing radiation on food products. Despite some public concern, the process is safe, free from chemical residues and presents advantages for preservation and storage. Nevertheless, safety dose parameters must be adopted in irradiation procedures to inhibit formation of undesirable and/or toxic products, for example, 2-ACB’s (2-alkylcyclobutanones) that are cyclic compounds containing four carbon rings that can be formed in food when its fat content is irradiated through breakdown of fatty acids. 2-ACBs are considered a unique class of compounds due to divergences between results of its mutagenicity potential collected from different studies. In this study, a cell population collected from rat livers were chosen for in vivo genotoxicity analysis because the importance of the liver in the metabolization of compounds. Analysis was performed using the micronuclei test using flow cytometry, allowing faster analysis, use of few materials and reduction in the number of animals, what is a subject much addressed currently in research. Irradiated rat diet did not show any genotoxic effect on liver cell populations. The improvement of the techniques is important for the future of the research since the irradiation process is already consolidated.
Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2
2017 - BARBEZAN, ANGELICA B.; SANTOS, CARLA J.B.; CARVALHO, LUMA R.; VIEIRA, DANIEL P.; SANTELLI, GLAUCIA M.M.; VILLAVICENCIO, ANNA L.C.H.
Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line.