Abstract / Summary
Objectives: Bentazone is a widely used selective herbicide frequently detected in environmental water sources, raising concerns regarding its potential effects on non-target organisms. Although various pesticides have been associated with male reproductive toxicity, the effects of bentazone on testicular cells remain insufficiently characterized. This study aimed to assess the cytotoxic and apoptotic effects of bentazone in TM3 Leydig and TM4 Sertoli cells, with a particular focus on identifying potential cell type-specific responses.Materials and Methods: Cell viability using 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay was assessed using spectrophotometry, and intracellular reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), and apoptotic and necrotic cell populations were analyzed using flow cytometry.Results: Bentazone exposure resulted in concentration-dependent cytotoxicity, with TM3 cells showing greater sensitivity than TM4 cells (IC: 202.6 µM vs 464 µM). A dose-dependent increase in apoptosis was found in TM3 cells without significant changes in ROS production or MMP, indicating a ROS-independent response. In contrast, TM4 cells exhibited increased ROS production without accompanying MMP loss, along with a biphasic apoptotic pattern characterized by higher apoptosis at lower concentrations and attenuation at higher doses.Conclusion: Bentazone exhibits cytotoxic and cell type-specific apoptotic effects in TM3 and TM4 cells, likely involving multiple non-overlapping mechanisms beyond classical oxidative stress pathways. These findings contribute to filling an important gap in the literature regarding bentazone's reproductive toxicity potential and provide a basis for future hypothesis-driven investigations into the underlying molecular mechanisms.