Materials and Methods: In this experiment, 10 adult and clinically healthy male Wistar rats weighing approximately 250-200 g were divided into two equal groups of control and treatment (n=5 in each). In the control group, normal saline and in the treatment group, 20% calcium chloride was injected into the testicular tissue. Blood samples were taken at the beginning and end of the experiment (day 21) and the superoxide dismutase (SOD), glutathione peroxidase (GPx), and testosterone levels were measured. Then, the tissue samples of testis were evaluated on days 7, 14, and 21.

Results: On day 21, SOD and GPx enzymes significantly increased in the treatment group compared to day zero (P = 0.01). However, calcium chloride injection reduced the mean testosterone levels on day 21 compared to day zero (P = 0.06). Meanwhile, the mean number of sperms in the right testis of treated mice significantly decreased (P = 0.03). Seminiferous tubule necrosis significantly increased on day 7, neutrophil infiltration significantly increased on days 7 and 14, and calcification significantly increased on days 7 and 21 (P < 0.01).

Conclusions: Although chemical sterilization using 20% calcium chloride can lead to testicular degeneration, it is not a recommended method because it does not reduce testosterone and activates oxidative stress biomarkers. However, the negative effects might be eliminated in future studies using some substances such as tannic acid.