Therapeutic potential of testosterone on sperm parameters and chromatin status in fresh and thawed normo and asthenozoospermic samples.

BACKGROUND: Hormonal changes alter the physiological level of ROS and cause oxidative stress in the cell. As estimated, hormonal deficiencies, environmental and ideological factors make up about 25% of male infertility. Pathogenic reactive oxygen species (ROS) is a chief cause of unexplained infertility. Limited studies exist on the effects of testosterone on human sperm culture. Therefore, in the current study, the effect of different doses of testosterone on sperm parameters and chromatin quality was investigated. MATERIALS AND METHODS: Semen samples from 15 normospermic and 15 asthenospermic patients were prepared by swim up method, and then were divided into four groups by exposing to different concentrations of testosterone (1, 10, and 100nM) for 45min. Samples without any intervention were considered as control group. All samples were washed twice. Sperm parameters and chromatin protamination were assessed in each group and the remains were frozen. After two weeks, all tests were repeated for sperm thawed. Also, the MSOM technique was used to determine the sperm morphology of class 1. RESULTS: Although sperm parameters were not show any significant differences in normospermic and asthenospermic samples exposed to different concentrations of testosterone before and after freezing, chromatin protamination was significantly decreased in the normospermic samples exposed to 10nM of testosterone before freezing (p<0.006), as well as 1 and 10nM of testosterone after freezing compared to control samples (p=0.001 and p=0.0009, respectively). Similarly, chromatin protamination in the asthenospermic samples was significantly decreased at concentration of 1nM of testosterone before and after freezing (p=0.0014 and p=0.0004, respectively), and at concentration of 10nM of testosterone before and after freezing (p=0.0009, p=0.0007) compared to control samples. CONCLUSION: Using a low dose of testosterone in the sperm culture medium, has positive effects on chromatin quality.

Effect of Nanoalumina on Sex Hormones and Fetuses in Pregnant Rats.

OBJECTIVE: This study aimed at investigating the effect of nanoalumina on sex hormones, and fetuses in pregnant rats. METHODS: In this study, sixty-four pregnant rats were divided into eight groups. The control and the injection-control group received normal food and water, and 0.5 ml of distilled water, respectively. Treatment groups were treated with 25, 50, 100, 250, 500, and 1000µg/ml concentrations of Nanoalumina from the 7th day until the 18th day of pregnancy. On the 18th day, the rats were investigated in terms of their hormone levels. We evaluated the number of healthy and aborted offspring, as well as fetus size. RESULTS: Nanoalumina caused an increase in progesterone hormones at the concentrations of 250, and 500µg/ml, and a significant reduction in estrogen hormone and aborted fetuses at the concentrations of 250 and 500µg/ml (p<0.05). The largest and smallest size of fetuses were observed in 500µg/ml and 1000µg/ml, respectively. The highest number of aborted fetuses was observed in the group treated with the 500µg/ml concentration. There was no aborted fetuses with 25, 50,100, control, and injection-control groups. CONCLUSIONS: Due to nanoalumina toxicity, it must be used with caution.

Does combination of estradiol and sesame oil improve the oocyte quality, embryo development and expressions of Zp3, E-cad, and Ctnnb1 genes in mice? An experimental study.

BACKGROUND: Aging may reduce oocyte maturation, embryo quality, and fertility potential. OBJECTIVE: To compare the effect of estradiol (E2) and sesame oil on oocyte and embryo quality between young and old mice. MATERIALS AND METHODS: Sixty old and young female mice were divided in to two groups (30 mice/group, grouped by age). Each group was divided into three subgroups of mice treated with sesame oil, E2 + sesame oil, and normal saline as control group. After ovulation induction, some oocytes were considered for in vitro fertilization and the rest were assessed for morphological status. After obtaining the two-cell embryos, the embryos were collected to determine the expression of zona pellucida (ZP) glycoprotein 3, E-cadherin, and ß-catenin genes and some of them followed until the blastocysts stage to evaluate the viability. RESULTS: The findings showed that the mean ZP and perivitelline space thickness increased in the old mice that received the E2 + sesame oil treatment. The number of 2-cell embryos, blastocysts, and live cells were significantly higher in the old group treated with sesame oil respectively (p = 0.018, 0.002, and < 0.0001, respectively). The normal ZP shape and refractile body numbers increased in the old mice that were treated with sesame oil, respectively. The E-cadherin gene was downregulated in the treatment groups compared to the controls. CONCLUSION: Sesame oil showed a better response in the old mice, because aging is associated with an increased rate of reactive oxygen species, causing deficiencies in both oocyte and embryo qualities.

Protective effect of vitamin E on sperm parameters, chromatin quality, and DNA fragmentation in mice treated with different doses of ethanol: An experimental study.

BACKGROUND: Excessive consumption of alcohol induces an increase in oxidative stress production and can lead to detrimental effects on the male reproductive system. OBJECTIVE: To evaluate the possible protective effects of coadministration of vitamin (vit) E on the detrimental changes in the sperm quality of mice administered ethanol. MATERIALS AND METHODS: Fifty-four BALB/c mice were categorized into nine groups (n = 6/each). The control group received a basal diet while the eight experimental groups received ethanol 10%; ethanol 20%; vit. E 100 mg; vit. E 200 mg; ethanol 10% + vit. E 100 mg; ethanol 10% + vit. E 200 mg; ethanol 20% + vit. E 100 mg; ethanol 20% + vit. E 200 mg. After 35 days, the sperm parameters and sperm chromatin were assessed. RESULTS: The results demonstrated a significant reduction in the motility rate, normal morphology rate, viability rate, increase in abnormal DNA structure and packaging (TB staining), and DNA damage (TUNEL) in ethanol consumer groups. In addition, the findings showed a significant increase in the aforementioned parameters in ethanol- and vit. E-consumer groups compared to the ethanol-only consumer groups. The ethanol group received 20% of the most damage among the groups. The group receiving vit. E 100 mg and those receiving ethanol 10% + vit. E 200 mg gained the highest benefit among the groups. CONCLUSION: Sperm forward progressive motility, normal morphology rate, and viability decreased in the ethanol groups. Also, the rates of spermatozoa with abnormal DNA structure and DNA fragmentation increased in the ethanol groups. Our findings revealed that the coadministration of vit. E and ethanol can protect destructive changes in DNA structure and damage.

The effects of sesame oil and different doses of estradiol on testicular structure, sperm parameters, and chromatin integrity in old mice.

OBJECTIVE: Studies of the effects of estrogens on the male reproductive system have emphasized the role of these hormones in male fertility. Sesame oil has many phytoestrogenic compounds and may improve male fertility. This study investigated the effects of sesame oil and different concentrations of estrogen on sperm parameters and DNA integrity in male mice. METHODS: Twenty old NMRI (The Naval Medical Research Institute) male mice (40 weeks; weight, 30-35 g) were treated with sesame oil or different concentrations of estrogen (estradiol, 1 and 10 µL/kg/ day) or received no treatment (controls). After 35 days, sperm parameters and DNA integrity were assessed and analyzed. RESULTS: Sperm count, progressive motility, and morphology were decreased in the group that received 10 µL/kg of estradiol. A remarkably lower percentage of DNA fragmentation and protamine deficiency were detected in the group that received 1 µL/kg of estradiol. In the groups that received sesame oil and 1 µL/kg of estradiol, the numbers of spermatogonia and Leydig cells were higher than in controls. The combination of sesame oil and 1 µL/kg of estradiol led to improved sperm parameters and chromatin and testicular structure. CONCLUSION: Based on this study, consumption of sesame oil and a low concentration of estradiol may improve testicular function in older mice.

The effect of testosterone and antioxidants nanoliposomes on gene expressions and sperm parameters in asthenospermic individuals.

BACKGROUND: Antioxidants that used for the infertility treatment cannot have their complete effectiveness, because of their instability in the culture medium. SIGNIFICANCE: One of the most advances, in the drug delivery systems, is nanoliposomes-loaded, as biodegradable and bioavailable carriers. Hormonal and antioxidant agents encapsulating inside the nanoliposomes were used, to increase the effectiveness of antioxidants in the sperm culture medium. MATERIALS: Semen sample from 15 asthenospermia were divided into 10 equal parts. After preparation, the sperms were incubated with free form of drugs and nanocarriers contained resveratrol, catalase, resveratrol-catalase and testosterone for 45 min. All sperm parameters, sperm DNA and gene expressions were evaluated before and after freezing. RESULTS: Before freezing, all nanocarriers and free testosterone showed higher sperm motility compared to free drugs (p=.000). Free Testosterone and free resveratrol-catalase had higher DNA damage compared to nanocarriers (p=.000). Before freezing, the blank nanoliposome and testosterone nanoliposomes had the lowest HSP70 gene expression respectively (p = 0.005) (p = 0.001). After freezing, a significant reduction in sperm motility was observed in the free resveratrol-catalase group (p=.003). Also, a significant increase in sperm viability was observed in the free testosterone and nanoliposomes of blank and testosterone (p > 0.05). The least DNA fragmentation was related to catalase nanoliposomes (p=.000). All nanoliposomes, especially catalase, had the highest percentage of class I morphology compared to the control group (p=.000). CONCLUSIONS: Nanoliposomes could improve the sperm parameters and DNA integrity before and after freezing, by increasing the effectiveness of antioxidants. So, it can be recommended in the ART lab.

Does resveratrol affect prepared sperm parameters and chromatin quality in normozoospermic and asthenozoospermic patients before and after freezing? A lab trial study.

BACKGROUND: Previous studies have examined the effect of resveratrol as a potent antioxidant for free radicals in semen. While, the prepared spermatozoa are more affected by ROS factors due to centrifugation and incubation. OBJECTIVE: To evaluate the RSV's effects on the prepared sperm parameters and chromatin quality in both normozoospermic and asthenozoospermic cases before and after freezing. MATERIALS AND METHODS: The sample of 10 normozoospermic and asthenozoospermic men was prepared through the swim-up method. The groups were then divided into two samples of control and experimental (exposure to 30 µmol/l of RSV) to evaluate and compare the sperm parameters and chromatin quality before and after freezing. RESULTS: The motility and viability of spermatozoa were seen to be significantly different before and after freezing separately in the control and treatment samples of the groups (p ≤ 0.001 and p = 0.001, respectively). However, the stated difference between the control and treatment samples of normozoospermic and asthenozoospermic patients were not significant (p > 0.05). In addition, the sperm morphology and chromatin quality were not significantly different between the two samples of each group; nonetheless, chromatin quality of the treated sample was better than that of the control before and after freezing. CONCLUSION: Despite the protective effects of RSV on the semen samples, RSV cannot affect significantly the prepared sperms parameters and chromatin quality in normozoospermic and asthenozoospermic patients.

Medium containing different concentrations of catalase as a strategy for optimising sperm parameters and chromatin in normospermic persons.

The aim of this study was to comprise the effect of catalase on sperm parameters and chromatin in normospermic persons. Semen samples were obtained from fertile men. A certain amount of different concentrations of catalase (0.1, 1, 10, 50, 100, 150 and 200 IU.ml) was added to each vial containing semen. Control group had similar condition to treated groups without treatment. Treatment was done for one hour in incubator and 4 and 24 hr in room temperature. Sperm parameters (motility, viability and morphology) and chromatin were evaluated after incubation. The results show that percentage of motility was insignificantly increased at concentration of 100 IU.ml catalase. This increase was higher than other examined concentration in all incubation time. The increase in sperm motility had significant difference in concentrations of 100 IU.ml with other concentrations. Other parameters showed no significant difference in all concentrations. Regarding the health of sperm chromatin, low concentrations of catalase had significant effect on this variable. This effect was more in low concentrations than high concentrations. This study showed the use of lower concentrations of antioxidant can improve the sperm parameters and chromatin quality. The low concentrations of catalase led to protection of chromatin and optimisation of sperm parameters.