Assessment of human sperm function after hydrogen peroxide exposure. development of a vaginal contraceptive.
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abstract
The reactive oxygen species (ROS) that is most damaging to human spermatozoa is hydrogen peroxide. Using an artificial medium Ham's F-10, we have evaluated the effect of different concentrations of hydrogen peroxide (H(2)O(2)) on various sperm function characteristics to develop a water-based vaginal contraceptive. H(2)O(2) at 30 and 60 micro M had no effect on sperm motility. At 120 micro M of H(2)O(2), a significant reduction (90-95%) in motility was observed after 120 min. However, at 600 micro M of H(2)O(2), all the sperm were found immotile within 15 min of incubation. At much higher concentrations (1200-1500 micro M), the effect was immediate. After immobilization with H(2)O(2) (600 micro M in media), viability in sperm declined to 36% after 30 min, and was further reduced to 5% in 60 min. On the other hand, hypo-osmotic swelling response in these sperm went down drastically from 72 to 46% immediately after immobilization and there was no response at all after 60 min. In contrast, when Triton-x-100 (0.01%), a known spermicidal agent, was used in place of H(2)O(2), the immediate loss of sperm motility corresponded simultaneously with the loss of the other two functional parameters. Further, H(2)O(2) immobilized sperm revealed reduced (25-30%) nuclear chromatin decondensation as compared to 70% for the controls. A composition containing 2% H(2)O(2) in 0.9% NaCl was tested in rats as a vaginal contraceptive, depicting 100% efficacy in mating studies after 2 h of vaginal application. The findings indicate the potential of developing a water-based vaginal contraceptive using H(2)O(2) at an optimal concentration as utilized in the present study.
