Surrogate post-coital testing for contraceptive efficacy against human sperm activity in the ovine vaginal model†

High unintended pregnancy rates are partially due to lack of effective nonhormonal contraceptives; development of safe, effective topical vaginal methods will address this need. Preclinical product safety and efficacy assessment requires in vivo testing in appropriate models. The sheep is a good model for the evaluation of vaginally delivered products due to its close similarities to humans. The study objective was to develop an ovine model for efficacy testing of female nonhormonal contraceptives that target human sperm. Fresh human semen was pooled from male volunteers. Nonpregnant female Merino sheep were treated with control or vaginal contraceptive product (IgG antibody with action against sperm or nonoxynol-9 [N9]). Pooled semen was added to the sheep vagina and mixed with product and vaginal secretions. Microscopic assessment of samples was performed immediately and progressive motility (PM) of sperm was compared between treatments. Cytokines CXCL8 and IL1B were assessed in vaginal fluid after instillation of human semen. No adverse reactions or elevations in proinflammatory cytokines occurred in response to human semen. N9 produced signs of acute cellular toxicity while there were no cellular changes after IgG treatment. N9 and IgG had dose-related effects with the highest dose achieving complete sperm immobilization (no sperm with PM). Surrogate post-coital testing of vaginally administered contraceptives that target human semen was developed in an ovine model established for vaginal product preclinical testing. This expanded model can aid the development of much needed nonhormonal topical vaginal contraceptives, providing opportunities for rapid iterative drug development prior to costly, time-intensive human testing.

Sperm impairing microbial factor: potential candidate for male contraception

Background Despite significant advances in contraceptive options for women, vasectomy and condoms are the only options available for male contraception. Due to this limitation, the burden of contraception resides on the shoulders of females only. Therefore, there is an urgent need to develop a safe, effective and reversible method of contraception for men. Amongst the alternative approaches, microbial derived products are gaining attention of the scientific world to combat unintended pregnancies. Earlier in our laboratory, sperm impairing microbial factor (Sperm immobilization factor) isolated from Staphylococcus aureus has shown excellent contraceptive efficacy in female mice. Keeping this in mind, the present study was carried out to exploit the sperm immobilization factor (SIF) as potential male contraceptive using vas deferens for administration in mouse model. Methods SIF (10, 50, 100 or 200 μg) was inoculated in the lumen of right vas deferens whereas the left vas deferens served as control. The mice were sacrificed at Day 3, 7, 14, 21, 30, 45, 60 and 90 after inoculation and the results in terms of change in body weight, seminal parameters, Tissue somatic indices (TSI), haematological parameters, serum level of testosterone, lipid peroxidation and histology were studied. In order to ratify the SIF induced azoospermia SIF (200 μg) was administered with different doses viz. 100, 200, 300, 400 or 500 μg of SIF binding receptor extracted from mouse spermatozoa. Results The weight profile studies of all the experimental groups showed no significant change in the initial and final body weight. In case of seminal parameters, the results revealed that right vas deferens treated with SIF showed azoospermia and with 200 μg of SIF it persisted up to 90 days. TSI of reproductive organs and non-reproductive organs showed no significant change in all the experimental groups. The haematological indices were found to be unaltered throughout the course of investigation however significant decrease in testosterone level was observed in the treated mice. The treatment also affected the oxidative status of the testis. Further, histological studies revealed hypospermatogenesis and late maturation arrest on treated side whereas the left side which served as control showed normal tissue histology. SIF induced azoospermia was ameliorated when administered with 400 μg of SIF binding receptor from mouse spermatozoa. Conclusion SIF, when administered via intra vas deferens route, could lead to complete azoospermia. Therefore, it could be considered as a potential male contraceptive.

Effects of different polyvinylpyrrolidone concentrations on intracytoplasmic sperm injection

SummaryTo explore whether different polyvinylpyrrolidone (PVP) concentrations affect the results of intracytoplasmic sperm injection (ICSI), a prospective study was conducted for 194 couples undergoing 210 ICSI therapy cycles. These cycles were divided into three groups (10, 7 and 5% groups) using the corresponding concentration of PVP for sperm immobilization. The main outcome measures were analyzed. Results indicated that, with a decrease in PVP concentrations, all of the main outcome measures increased. In particular, the high-quality cleavage embryo rate in the 7% group was significantly lower than in the 5% group (P < 0.01), and the cleavage, high-quality cleavage embryo, and high-quality blastocyst rates in the 5% group were significantly higher than those in the 10% group (all P < 0.001). For high-/intermediate-quality semen, all of the main outcome measures were significantly increased with 5% PVP. For the poor-quality semen, only the high-quality cleavage embryo and high-quality blastocyst rates were significantly higher in the 5% group. Therefore, lowering PVP concentrations greatly promoted the development of embryos in ICSI cycles, with an optimal concentration of 5% for ICSI.

Shared Antigenic Determinants Between Spermatozoa and Bacteria: An Experimental Study

Sperm immobilization factor (SIF), the secretory protein of Staphylococcus aureus, is known to cause complete immobilization, death and morphological alterations in mouse spermatozoa in vitro. However, the present study aims to explore a newer dimension of SIF i.e., to bind to motile and non-motile bacteria and its ability to induce immobilization of motile bacteria in vitro. The results showed that 800µg of SIF caused complete immobilization of motile bacteria, however, death and morphological alterations could not be observed even with 1000µg of SIF. Furthermore, this SIF-mediated bacterial immobilization was reversed when each of the SIF-binding receptor from mouse spermatozoa and bacteria (Escherichia coli and Streptococcus pyogenes) was incubated with bacteria, thereby, providing an experimental evidence of similarity between the antigenic determinants present on spermatozoa and bacteria against a common ligand, SIF.

Spermicidal Constituents of Ethanolic Extract of Sacoglottis gabonensis Stem Bark

Aim: To isolate the spermicidal constituents of Sacoglottis gabonensis. Materials and methods: The ethanolic extract with partitioned fractions of Sacoglottis gabonensis stem bark were subjected to sperm immobilization assay. The most active EtOAc fraction was further purifi ed by column and Semi-Preparative High Performance Liquid Chromatography to give compounds which were characterized by spectroscopic methods (UV, LC/MS, and NMR). The compound(s) was also tested for sperm immobilization activity. Results: The ethanolic extract showed 100% signifi cant (p < 0.05) sperm immobilization activity at a concentration of 30 mg/mL at 20 s compared to both negative and positive controls. The most active ethyl acetate fraction yielded methyl 3,5-dihydroxy-4-methoxybenzoate, eriodictyol and bergenin. Bergenin had 100% sperm immobilization activity at 20 mg/mL in 60 s which was signifi cant (p < 0.05) also when compared to the positive and negative control while methyl 3,5-dihydroxy- 4-methoxybenzoate, eriodictyol were not active. Conclusion: The active spermicidal constituent in Sacoglottis gabonensis stem bark extract is bergenin. However, methyl 3,5-dihydroxy-4-methoxybenzoate and eriodictyol showed no activity. This plant is known for its aphrodisiac action; hence, caution may have to be exercised in its use because of its spermicidal eff ect.