Effects of multiple doses of gonadotropin-releasing hormone agonist on the luteal-phase support in assisted reproductive cycles: A clinical trial study

Background: The effect of adding gonadotropin-releasing hormone (GnRH) agonist on the luteal phase support in assisted reproductive technique (ART) cycles is controversial. Objective: To determine the effects of adding multiple doses of GnRH agonist to the routine luteal phase support on ART cycle outcomes. Materials and Methods: This clinical trial study included 200 participants who underwent the antagonist protocol at the Research and Clinical Center for Infertility, Yazd, Iran, between January and March 2020. Of the 200, 168 cases who met the inclusion criteria were equally divided into two groups – the case and the control groups. Both groups received progesterone in the luteal phase, following which the case group received GnRH agonist subcutaneously (0/1 mg triptorelin) zero, three, and six days after the fresh embryo transfer, while the control group did not receive anything. Finally, chemical and clinical pregnancy rates, number of mature oocytes, fertilization rate, total dose of gonadotropin, and the estradiol level were determined. Results: The baseline characteristics were similar in both groups. No significant difference was observed between embryo transfer cycles. Clinical results showed that differences between the fertilization rate, chemical and clinical pregnancies were not significant. Conclusion: The results showed that receiving multiple doses of GnRH agonist in the luteal phase of ART cycles neither improves embryo implantation nor the pregnancy rates; therefore, further studies are required. Key words: Luteal phase, GnRH agonist, ART, Pregnancy rate

Is there any correlation between Estradiol supplementation, as luteal phase support, and clinical pregnancy in ART cycles? A cross-sectional study

Background: Endometrial receptivity is one of the important factors in assisted reproductive technology (ART) success. In the luteal phase of an ART cycle, serum estradiol (E2) and progesterone are often placed in low levels. Supporting the luteal phase with progesterone is a usual method. Objective: To evaluate the effects of E2 supplementation plus progesterone on the luteal phase support in the antagonist protocol who have undergone intracytoplasmic sperm injection-embryo transfer cycles. Materials and Methods: In this cross-sectional study, 200 patients with antagonist stimulation protocol, who had undergone intracytoplasmic sperm injection treatment, were divided into two groups based on the use of E2 supplementation. In both groups, 400 mg progesterone suppositories (Cyclogest®), twice a day/vaginally, was administered starting from the day of oocyte collection until the fetal heart activity. However, in the E2 group, in addition to progesterone, 4 mg tablet of E2 was received daily. Beta hCG was checked 14 days after the embryo transfer, and the clinical pregnancy rate was the main endpoint. Results: The patients’ characteristics were matched, and insignificant differences were observed, except for endometrial thickness. The clinical outcomes showed the rate of pregnancy was higher in the E2 group compared to the control group; nonetheless, statistically, there was no noticeable difference. Conclusion: E2 supplementation had no beneficial effect in the luteal phase support of IVF cycles. Nevertheless, more studies are required to confirm the supportive role of E2 supplementation for embryo implantation and to improve the outcomes in ART cycles. Key words: Luteal phase, Estradiol, Progesterone, ICSI, Pregnancy rate

Comparison of zeta potential and physiological intracytoplasmic sperm injection in obtaining sperms with a lower DNA fragmentation index: A cross-sectional study

Background: The sperm DNA fragmentation index (DFI) is one of the men’s reproductive health criteria that affects assisted reproductive technique outcomes. Efforts in obtaining high-quality mature sperms seem to be necessary. Advanced sperm selection techniques (including physiological intracytoplasmic sperm injection [PICSI], zeta potential, microfluidic, etc.) have gained popularity in this regard. Objective: The study aimed to compare the efficacy of zeta potential and PICSI sperm selection in obtaining sperms with better DNA integrity. Materials and Methods: In this cross-sectional study, 48 couples were enrolled where the male partner had increased sperm DFI in his ejaculated sample and the female was in normal reproductive health. For each male partner, the semen sample was processed with zeta potential and PICSI techniques, then the sperm DFI of neat semen was compared to zeta and PICSI samples by the sperm chromatin dispersion test. Results: Data showed that both the zeta potential and PICSI technique decreased sperm DFI in comparison with the neat semen sample (p < 0.001 for both). In addition, there was a statistically significant difference in sperm DFI between the PICSI and zeta potential samples (p < 0.01). Conclusion: The current study showed that both zeta potential and PICSI could result in sperm with a lower DFI. However, PICSI seems to be superior to zeta potential in this regard. Key words: PICSI, ZETA potential, Hyaluronic acid, DNA integrity

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. Key words: Resveratrol, Chromatin, Motility, Spermatozoa, Freeze

Effects of in-vitro application of pentoxifylline on the morphology of human spermatozoa after vitrification in asthenozoospermic patients

Cryopreservation of human spermatozoa is widely used in many assisted reproduction units to preserve male fertility [1]. Vitrification is based on the ultrarapid freezing and is routinely assayed for cryopreservation in assisted reproductive technology. Mohamed [2] showed that cryopreservation significantly affects progressive motility, viability and mitochondrial membrane potential of spermatozoa. Pentoxifylline (PX) is a phosphodiesterase considered to be a sperm movement enhancer, hyperactivation agent, inhibitor of reactive oxygen species and acrosome reaction-improving agent. The aim of our study was to evaluate the effect of in-vitro application of PX on sperm parameters and ultrastructure after vitrification. A total of 30 asthenozoospermic semen samples were selected and divided into two groups after vitrification: control (without PX) and experimental (with PX). A significant decrease in sperm motility, morphology and viability was observed post vitrification, but sperm motility was increased significantly following application of PX. On the other hand, PX did not exert any significant effect on the ultrastructure of the acrosome, plasma membrane and tail of vitrified spermatozoa

Does body mass index compromise assisted reproductive technique outcomes? A cross-sectional study

Background: Overweight and obese people face several health problems. Female obesity has been shown to reduce fertility in the general population. Assisted reproductive technology outcomes in obese cases are widely studied, but the results are inconclusive. Objective: This study aimed to compare live birth rate (LBR) among women with 4 different types of body mass index (BMI). Materials and Methods: In this cross-sectional study, data of 1611 women, who were candidates for fresh and frozen embryo transfer cycles, was extracted from 2051 medical files at the Reproductive Sciences Institute, Yazd, Iran from May 2019-May 2021. The participants were divided into 4 groups (underweight, normal, overweight, and obese) according to their BMI, and LBR was considered to be the main outcome. Results: Of 1611 women, 39 were underweight, 585 were normal, 676 were overweight, and 311 were obese. Underweight women had the lowest LBR (12.8%), but there was no statistically significant difference (p = 0.55). In addition, LBR was compared in the 4 BMI groups according to age, type of transfer cycle (fresh or freeze), and cause of infertility, and there was comparable LBR in the 4 BMI groups. However, metaphase 2 oocyte rate, doses of gonadotropin usage in the cycles, and estradiol level had statistically significant differences (p < 0.001). Conclusion: According to our study, obesity does not affect LBR in the IVF cycle, regardless of fresh or frozen embryo transfer cycles, different age groups, and causes of infertility. Key words: Body mass index, Assisted reproductive technique, Pregnancy outcome, Live birth rate, Age

Short and long term effects of different doses of paracetamol on sperm parameters and DNA integrity in mice

The aim was to survey the impact of normal and high doses of paracetamol consumption on sperm parameters and DNA integrity in mice. A total of 36 adult male mice were divided into three groups: mice of group A served as control fed on basal diet, group B received normal dosage of Paracetamol (66 mg kg/day) and basal diet, group C received high dosage of Paracetamol (100 mg kg/day) and basal diet for 35, 70 and 105 days. The cauda epididymitis of each mouse was dissected and placed in 1 ml of pre-warm Ham’s F10 culture medium for 20 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. Sperm chromatin quality was evaluated by chromomycin A3 staining (CMA3), aniline blue staining and sperm chromatin dispersion test (SCD). The results showed that almost all of the sperm parameters significantly decrease following consumption of normal and high dosage of Paracetamol in three periods of experiments in mice (p < 0.05). Regarding to SCD test, we found a highly significant difference only in dose effect, but in CMA3 test and aniline blue staining there was a significant difference (p < 0.05) in both dose and time effects. According to our results, paracetamol as an analgesic and antipyretic may have detrimental effects on sperm parameters and DNA integrity in mice

Effects of experimentally-induced diabetes on sperm parameters and chromatin quality in mice

Background: Diabetes mellitus (DM), primary or idiopathic is a chronic disorder of the carbohydrate, lipid and protein metabolism. DM may impact male reproductive function at several levels. It is shown that DM has detrimental effects on sperm parameters in human and experimental animals. Objective: The aim of this study was to observe the effects of diabetes on sperm parameters (viability, count, morphology and motility) and evaluation of sperm chromatin quality in mice. Materials and Methods: Totally twenty adult male Syrian mice were divided randomly into 2 groups (n=10). The animals of group A were considered as controls while group B mice were diabetic that received a single dose (200 mg/kg) streptozotocin (STZ) intra peritoneally. After 35 days, the cauda epididymis of each diabetic mouse was dissected and placed in culture medium for 30 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. The sperm chromatin quality and DNA integrity, was evaluated with Aniline Blue (AB), Toluidine blue (TB), Acridine orange (AO) and Chromomycin A3 (CMA3) staining. Results: In sperm analysis, the diabetic mice had poor parameters in comparison with control animals (p=0.000). Regarding sperm chromatin quality, the results of TB and AO tests showed statically significant differences between two groups, but in AB and CMA3 staining, we didn’t see any differences between them. Conclusion: The results showed that STZ-induced diabetes mellitus may influence the male fertility potential via affecting sperm parameters and DNA integrity in mice. However, according to our data, the diabetes doesn’t have any detrimental effects on histone-protamines replacement during the testicular phase of sperm chromatin packaging

Effects of Multiple Doses of Gonadotropin-releasing Hormone Agonist on the Luteal-phase Support in Assisted Reproductive Cycles: A Clinical Trial Study

Background: The effect of adding gonadotropin-releasing hormone (GnRH) agonist on the luteal phase support in assisted reproductive technique (ART) cycles is controversial. Objective: To determine the effects of adding multiple doses of GnRH agonist to the routine luteal phase support on ART cycle outcomes. Materials and Methods: This clinical trial study included 200 participants who underwent the antagonist protocol at the Research and Clinical Center for Infertility, Yazd, Iran, between January and March 2020. Of the 200, 168 cases who met the inclusion criteria were equally divided into two groups – the case and the control groups. Both groups received progesterone in the luteal phase, following which the case group received GnRH agonist subcutaneously (0/1 mg triptorelin) zero, three, and six days after the fresh embryo transfer, while the control group did not receive anything. Finally, chemical and clinical pregnancy rates, number of mature oocytes, fertilization rate, total dose of gonadotropin, and the estradiol level were determined. Results: The baseline characteristics were similar in both groups. No significant difference was observed between embryo transfer cycles. Clinical results showed that differences between the fertilization rate, chemical and clinical pregnancies were not significant. Conclusion: The results showed that receiving multiple doses of GnRH agonist in the luteal phase of ART cycles neither improves embryo implantation nor the pregnancy rates; therefore, further studies are required. Key words: Luteal phase, GnRH agonist, ART, Pregnancy rate