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  • Title: Dose-response effects of gramicidin-D, EDTA, and nonoxynol-9 on sperm motion parameters and acrosome status.
    Author: Centola GM.
    Journal: Contraception; 1998 Jul; 58(1):35-8. PubMed ID: 9743894.
    Abstract:
    Previous reports showed that gramicidin-D (G-D), a polypeptide with antiviral and antimicrobial properties, nonoxynol-9 (N9), a common spermicidal detergent, and EDTA, a Ca-Mg chelating agent, inhibited sperm motility and cervical mucus penetration. The purpose of this study was to determine the dose-response effects of G-D, N9, EDTA and G-D + EDTA on sperm motion parameters and acrosome status. Semen specimens from known fertile donors were subjected to computer-assisted semen analysis of motility, path velocity, progressive velocity, and hyperactivation prior to and after incubation with varying concentrations of gramicidin-D, EDTA and nonoxynol-9. Each specimen was also prepared for acrosome status using rhodamine isothiocyanate conjugated pisum sativum agglutinin (RITC-PSA). There was a significant decrease in motility by G-D, EDTA, G-D + EDTA, and N9 at all doses as compared to the fresh specimen. N9 completely immobilized all sperm at each dose. Progressive velocity and path velocity also decreased in a dose-response manner. Sperm hyperactive motility also significantly decreased in all groups. The majority of sperm remained acrosome intact following exposure to all doses tested, whereas N9 resulted in complete breakdown/release of the acrosomal contents. This study confirms previous reports that G-D, EDTA, and N9 significantly impair sperm motility and motion parameters. The effective 100% inhibitory concentration was seen only with N9, whereas G-D, EDTA, and G-D + EDTA resulted in incomplete impairment of sperm motion parameters. At the concentrations used, N9 demonstrated potent spermostatic activity. Gramicidin-D and EDTA should be further studied for their potential contraceptive spermostatic activity. Gramicidin-D (G-D), a polypeptide with antiviral and antimicrobial properties, the spermicidal detergent nonoxynol-9 (N-9), and the Ca-Mg chelating agent EDTA have been shown in previous studies to inhibit sperm motility and cervical mucus penetration. This study utilized computer-assisted methods to investigate the dose-response effects of incubation with G-D, N-9, EDTA, and G-D plus EDTA on sperm motion parameters and acrosome status. Semen specimens were acquired within 30 minutes of ejaculation from six fertile US sperm donors. Compared to the fresh (untreated) specimen, there was a significant decrease in sperm motility produced by G-D, EDTA, G-D plus EDTA, and N-9 at all doses. Progressive and path velocity and sperm hyperactive motility also decreased in a dose-response manner in all groups. However, sperm immobilization was complete at the concentrations used only with N-9. The majority of sperm remained acrosome-intact after exposure to all tested doses of G-D and EDTA, but N-9 resulted in complete breakdown and release of the acrosomal contents. A combination of N-9 and G-D or N-9 and EDTA at lower doses might produce the desired inhibition of sperm motility without toxicity and this possibility should be investigated. At the present time, however, G-D or EDTA, alone or in combination, cannot be considered effective contraceptive agents.
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