Evaluation of Cytotoxic Effects of 2-Monochloropropanediol (2-MCPD) on Mouse TM3 Leydig and TM4 Sertoli Cells in vitro

Abstract

Aim: 2-Monochloropropanediol (2-MCPD), 3-monochloropropanediol (3-MCPD), their esters and glycidyl fatty acid esters (GE) are important food contaminants originating mainly during food processing. Recently, the detection of these contaminants in different foods and vegetable oils, especially in palm oil, margarines, and processed foods such as pastries and cakes and in baby foods raises concerns about their safety. According to the European Food Safety Authority (EFSA) scientific report published in May 2016, foods and vegetable oil contaminated with 3-MCPD, 2-MCPD and their fatty acid esters and GE were reported as harmful to human health. However, limited data on 2-MCPD toxicity preclude a complete assessment of its toxicity. For this reason, the cytotoxic, apoptotic effects and deoxyribonucleic acid (DNA) damage potential of 2-MCPD were evaluated on TM3 Leydig and TM4 Sertoli cells.

Material and method: The cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) assays. Annexin V-FITC/PI double staining kit was used to indicate the type of cell death by flow cytometry. The alkaline comet assay was used to estimate the genotoxic effect.

Results: The cell viability assays showed that 2-MCPD was not cytotoxic within the tested concentrations up to 1000 µM on both cell lines. These sub-toxic concentrations of 2-MCPD (1-1000 µM) also did not induce DNA damage both on TM3 Leydig and TM4 Sertoli cells.

Conclusion: Although results demonstrate that tested concentrations of 2-MCPD (1-1000 µM) were neither cytotoxic nor apoptotic but may cause cell death and DNA damage at higher concentrations. On the other hand, those high concentrations are far from the daily exposure doses.