Effect of folic acid and zinc sulphate on endocrine parameters and seminal antioxidant level after varicocelectomy

Varicocele is among the most common problems which may lead to male infertility. Spermatogenesis is impaired as a consequence of this vascular defect, through mechanisms that are not well described. This study aimed to evaluate serum hormonal level (inhibin B, FSH and testosterone) and seminal plasma antioxidant defence levels after folic acid and zinc sulphate administration in varicocelectomised patients. Participants were randomly allocated to four experimental groups. Our randomisation schedule was as follows: zinc sulphate/ folic acid, folic acid, zinc sulphate and placebo. The patients underwent varicocelectomy, before which a blood and semen sample were obtained and also three and six months after varicocelectomy for evaluation of blood hormonal level (FSH, testosterone, inhibin B) and seminal oxidative stress status (nitric oxide, superoxide dismutase, total antioxidant capacity). Patients in different groups took orally one capsule per day after dinner following varicocelectomy for 6 months. A significant rise in peripheral blood inhibin B and seminal plasma activity was detected in the zinc sulphate/folic acid group after 6 months. The present clinical trial indicates a change in the hormonal status of varicocelectomised patients following long-term administration of zinc sulphate and folic acid

Recruiting testicular torsion introduces an azoospermic mouse model for spermatogonial stem cell transplantation

Purpose: To investigate the long-term effect of testicular torsion on sperm parameters and testis structure in order to introduce a novel mice azoospermic model for spermatogonial stem cell transplantation. Materials and Methods: Unilateral testicular torsion was created. The animals were divided into two groups each containing 15 mice. They underwent 2 and 4 hours of unilateral testicular ischemia, respectively. All animals in this experiment were aged matched. The experimental (n = 5) groups were studied 2, 4 and 10 weeks after testicular ischemia reperfusion. Moreover, the left testes and epididymis were removed for sperm analysis and for weight and histopathological evaluation. Finally isolated spermatogonial stem cells were transplanted in the testes that underwent 2 hours of ischemia reperfusion, two weeks post-surgery. Results: All the investigated parameters demonstrated a sharp decline at 2, 4 and 10 weeks after testicular torsion, whereas 2-hour ischemia was found to be less injurious in testicular tissue structure. Two months after xenotransplantation, the transplanted cells were localized in the basal of the seminiferous tubules of the recipient ischemic testes. Conclusion: Torsion can cause permanent azoospermia in mouse. Also Testicular torsion 2 weeks after the 2 hours ischemia reperfusion may prove useful for recipient preparation for SSCs transplantation in mouse

The development of mouse early embryos in vitro in fibroblasts and cumulus cells co-cultures supplemented with retinoic acid

This study was designed to examine the effects of retinoic acid adding to cumulus and/or fibroblast cells monolayer on the development of mouse early embryos. One-cell mouse embryos were obtained from NMRI mice after superovulation by an intraperitoneal injection of 5 IU equine chorionic gonadotrophin (eCG) followed 48 hrs later by 5 IU human chorionic gonadotrophin (hCG). Mouse embryonic fibroblasts (MEF) were obtained from mouse fetuses and cumulus cells (CC) were prepared from mouse cumulus-oocyte complexes (COCs). To produce monolayer of cumulus and fibroblast cells, 1.0 × 105 cells/ml were plated into culture dishes in 100 μl droplets. The collected mouse embryos were cultured randomly into six different conditions, being supplemented (experiment, Exp) or not (control, Con) with 0.28 μg/ml of retinol acetate methyl-β-cyclodextrin (RA) for 96 hrs at 37°C in 5% CO2 in air, including: (1) culture media only (Con 1); (2) culture media plus RA (Exp 1); (3) co-culture with CC (Con 2); (4) co-culture with CC plus RA (Exp 2); (5) co-culture with MEF (Con 3) and (6) co-culture with MEF plus RA (Exp 3). The culture medium was Alpha Modification of Minimum Essential Medium Eagle (α-MEM) + 10% fetal bovine serum (FBS) with 100 IU/ml penicillin and 100 μg/ml streptomycin. The proportions of embryos passing the two-cell block were significantly higher in the MEF (Con 3) group compared to the other treatment groups (P<0.05). The percentage of the two-cell passed embryos developing to the blastocyst stage was significantly higher in the co-culture groups than that of the culture medium alone (P<0.05). After 96 hrs in culture, the rate of blastocyst stage for both groups of CC co-culture treatment (Con 2 and Exp 2) was identical but, adding RA into the MEF co-culture (Exp 3) resulted significantly lower in vitro development than that of the Con 3 group (29.2% vs. 57.7%, P<0.05). These results suggest that supplementation of co-culture groups with RA could not affect the embryos passing the block and developing to the blastocyst stage, although the presence of RA into the culture medium alone may improve passing the critical two-cell stage. Also, in vitro addition of RA to cells without receptors for retinol during long term co-culture may result early embryonic growth retardation

Effects of concurrent chronic administration of alcohol and nicotine on rat sperm parameters

The prevalence of cigarette and alcohol consumption is high among young adult males during the reproductive period. The current study aimed to evaluate the impact of concurrent chronic administration of nicotine and ethanol on the quality of sperm in the rat. Fifty healthy Wistar male rats were randomly divided into five groups (n = 10) and were given the following for a period of 50 days: ethanol (E), nicotine (N), ethanol and nicotine (E/N); the control group (C) and an intact (I) group. Body weight as well as the weight, volume and dimensions of the testes and the weight of the cauda epididymidis and vas deference were measured. The concentration, motility, viability and membrane integrity of sperm were also assessed. There were no significant differences between body weight and all testis parameters including weight, volume and dimensions. The concentration and motility of sperm in the E/N group was significantly reduced compared with the control group (P < 0.01). Nevertheless, only a marginally significant decrease in sperm viability was found in the E/N group compared with the control group. The study indicates that concurrent chronic administration of ethanol and nicotine may disturb male reproductive function

Alteration of spermatogenesis following spermatogonial stem cells transplantation in testicular torsion-detorsion mice

Purpose: Testicular ischemia is the main consequence of testicular torsion, in both clinical and experimental aspects. Preservation and auto-transplantation of spermatogonial stem cells (SSCs) could be a new treatment for infertility in testicular ischemia following testicular torsion. Methods: To apply the idea in this study, animals were randomly divided into four groups of control, sham, with torsion, and with torsion followed by transplantation (TT). Isolated SSCs from neonatal mice were cultured and identified by flow cytometry (C-KIT�, INTEGRIN β1 +) and RT-PCR (Reverse transcription polymerase chain reaction) for specific spermatogonial cell markers (Oct4, Gfrα-1, Plzf, Vasa, Itgα6, and Itgβ1). SSCs were transplanted upon a 2-h testicular torsion in the TT group. Cultured cells were transplanted into ischemia reperfusion testicle 2 weeks post-testicular torsion. Eight weeks after SSCs transplantation, the SSCs-transplanted testes and epididymis were removed for sperm analysis, weight and histopathological evaluation, and pre- and post-meiotic gene expression assessment by qRT-PCR. Results: Our findings indicated that all evaluated parameters (epididymal sperm profile, Johnsen score, Plzf, Gfrα-1, Scp-1, Tekt-1 expressions, and histopathological profile) were significantly decreased following testicular torsion (group 3) when compared to the control group (p � 0.05). However, all abovementioned parameters showed a significant increase/improvement in torsion-transplantation group compared to torsion group. However, these parameters in the TT group were significantly lower in the sham and control groups (p � 0.05). Conclusion: SSCs transplantation could up-regulate the expression of pre- and post-meiotic genes in testicular ischemia, which resulted in improvement of both testicular function and structure after testicular torsion. © 2016, Springer Science+Business Media New York