Cytotoxic Effects of Multiple Pesticides and their Mixtures on Caco-2 Cells Evaluated by Using MTT and Trypan Blue Assays
- 1Ramazzini Institute
Abstract
BACKGROUND: Pesticides are extensively used in agriculture, raising concerns about their potential impact on human health through dietary and environmental exposure. OBJECTIVES: This study evaluated the in vitro cytotoxicity of ten commonly used pesticides and their mixtures (lambda-cyhalothrin, cypermethrin, deltamethrin, tebuconazole, glyphosate, acetamiprid, cyprodinil, piperonyl butoxide, fluopyram, and imazalil) on human intestinal Caco-2 cells. METHODS: Cytotoxicity was assessed using the MTT assay, as a measure of metabolic activity, and the trypan blue exclusion test, as an indicator of cell membrane integrity. FINDINGS: Results showed that high concentrations (100 mg/L) of all pesticides significantly reduced cell viability and vitality. Notably, glyphosate and tebuconazole exhibited significant toxicity even at lower concentrations, respectively 0.1 mg/L and 10 mg/L. Combination treatments (Top 3 and Top 8 pesticide mixtures) retained the cytotoxic effects observed for individual compounds, showing additive (non-synergistic) effects. CONCLUSIONS: Overall, these findings indicate that certain pesticides-based herbicides can exert cytotoxic effects on intestinal cells even at relatively low concentrations and highlight the importance of using the component-based approach in mixture risk assessment for humans. This study was performed as part of the EU SPRINT (Sustainable Plant Protection Transition: A Global Health Approach) project.
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BibTeX
@misc{truzzi2026cytotoxic,
title = {{Cytotoxic Effects of Multiple Pesticides and their Mixtures on Caco-2 Cells Evaluated by Using MTT and Trypan Blue Assays}},
author = {F. Truzzi and E. Tibaldi and S. Dilloo and R. Noferini and D. Sgargi and S. Panzacchi and E. D'Amen and F. Gnudi and G. Dinelli and P. T. J. Scheepers and D. Mandrioli},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.23.753704},
url = {https://www.biorxiv.org/content/10.64898/2026.09.23.753704v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Cytotoxic Effects of Multiple Pesticides and their Mixtures on Caco-2 Cells Evaluated by Using MTT and Trypan Blue Assays
AU - F. Truzzi
AU - E. Tibaldi
AU - S. Dilloo
AU - R. Noferini
AU - D. Sgargi
AU - S. Panzacchi
AU - E. D'Amen
AU - F. Gnudi
AU - G. Dinelli
AU - P. T. J. Scheepers
AU - D. Mandrioli
PY - 2026
DA - 2026/09/23
PB - bioRxiv
DO - 10.64898/2026.09.23.753704
UR - https://www.biorxiv.org/content/10.64898/2026.09.23.753704v1
AB - BACKGROUND: Pesticides are extensively used in agriculture, raising concerns about their potential impact on human health through dietary and environmental exposure. OBJECTIVES: This study evaluated the in vitro cytotoxicity of ten commonly used pesticides and their mixtures (lambda-cyhalothrin, cypermethrin, deltamethrin, tebuconazole, glyphosate, acetamiprid, cyprodinil, piperonyl butoxide, fluopyram, and imazalil) on human intestinal Caco-2 cells. METHODS: Cytotoxicity was assessed using the MTT assay, as a measure of metabolic activity, and the trypan blue exclusion test, as an indicator of cell membrane integrity. FINDINGS: Results showed that high concentrations (100 mg/L) of all pesticides significantly reduced cell viability and vitality. Notably, glyphosate and tebuconazole exhibited significant toxicity even at lower concentrations, respectively 0.1 mg/L and 10 mg/L. Combination treatments (Top 3 and Top 8 pesticide mixtures) retained the cytotoxic effects observed for individual compounds, showing additive (non-synergistic) effects. CONCLUSIONS: Overall, these findings indicate that certain pesticides-based herbicides can exert cytotoxic effects on intestinal cells even at relatively low concentrations and highlight the importance of using the component-based approach in mixture risk assessment for humans. This study was performed as part of the EU SPRINT (Sustainable Plant Protection Transition: A Global Health Approach) project.
ER -