Investigating the Cytotoxic Effect of Chitosan Nanoparticles on Sertoli Cell Lines

Introduction: Chitosan is one of the most widely used polymeric compounds used for drug delivery in medical systems. Considering that the properties of materials may be affected by their size, there are concerns about chitosan nanoparticles (CNPs) entering various body systems. Therefore, this study was conducted to evaluate the cytotoxicity of CNPs on the Sertoli cell line as one of the sensitive cells of the reproductive system. Methods: In this experimental study, the cytotoxicity of CNPs was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method 24 hours and 48 hours after exposure. After calculating the lethal concentration 50, Sertoli cells were exposed to a concentration lower than the lethal concentration 50. Then, RNA extraction, complementary DNA synthesis, and quantitative calculation of the expression of apoptotic genes BAX, BCL2, and Caspase were performed using the real-time polymerase chain reaction technique. The data were analyzed using SPSS 24. The normality of the data was assessed using the Shapiro-Wilk test, while significant differences were evaluated with the analysis of variance and the Tukey test (P<0.05). Results: By drawing a sigmoid curve and examining the cell viability, the half-maximal inhibitory concentration (IC50) values for 24 hours and 48 hours were 158.5 μg/mL and 79.4 μg/mL, respectively. Furthermore, the effect of a concentration lower than IC50 (i.e., 70 μg/mL of CNPs) on the expression of apoptotic genes revealed that CNPs significantly increased (P<0.05) the expression of BAX and caspase-3 genes but significantly decreased the expression of the Bcl2 gene in Sertoli cells. Conclusion: Although CNPs are increasingly utilized in clinical systems, it seems that they probably have a cytotoxic effect, at least on reproductive cells, due to their nano-sized dimension. Accordingly, it is recommended that these NPs be used after further studies.