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Hepatotoxicity induced by the herbicides Diuron and 2,4-D in Zebrafish cells cultured in different models: monolayer (2D) and three-dimensional (3D)

Grant number: 24/03537-0
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2024
Effective date (End): August 31, 2027
Field of knowledge:Health Sciences - Pharmacy - Toxicological Analysis
Principal Investigator:Lilian Cristina Pereira
Grantee:Paloma Vitória Lima Peixoto
Host Institution: Faculdade de Ciências Agronômicas (FCA). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated research grant:22/03045-4 - Validation of in vitro cell systems to assess the toxicity of socially relevant environmental pollutants, AP.PNGP.PI


The use of the herbicides Diuron and 2,4-D is common in Brazilian crops. In order to guarantee the protection of human health and the environment, it is essential that the toxicological potential of these compounds, both in their commercial formulations and in their metabolites and active ingredients (AI), is properly verified. Experimental studies have the ability to predict the toxicity of environmental contaminants of emerging concern, helping to understand the biological mechanisms involved in the risk to human health and also to support regulatory decisions by health and environmental agencies. Currently, the use of alternative methods for assessing the toxicity of chemical substances is an international trend in response to the 3Rs policy in animal experimentation, which restricts the use of animals in research. Among the new alternative methodologies applied to toxicology are monolayer cell cultures (2D), three-dimensional cell cultures (3D) and 3D bioprinting, which can use different cell lines, such as zebrafish liver cells (ZFL lineage). Zebrafish liver cell culture is a methodology that can help to elucidate key mechanisms of environmental contaminant toxicity and assist in the construction of conceptual frameworks ("Adverse Outcome Pathways", AOPs) that inform the mode of action (MoA) and mechanisms of harmful action of these contaminants. The use of ZFL cells is growing in toxicological studies, as the Zebrafish model organism has 71% similarity and conservation of xenobiotic metabolism and genetic material compared to humans. This project, which brings us into line with the worldwide trend of using alternative methods to assess the risk of chemical compounds, aims to assess the toxicity of the herbicides Diuron (and its metabolites DCA and DCPMU) and 2,4-D (both its acid IA 2,4-D and the commercial formula 2,4-D DMA). The experimental systems indicated - cell culture in 2D, 3D and 3D bioprinting - will provide scientific information that will support the implementation of coherent and effective strategies for controlling these environmental contaminants.

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