Relationship of sperm motility with clinical outcome of percutaneous epididymal sperm aspiration–intracytoplasmic sperm injection in infertile males with congenital domestic absence of vas deferens: a retrospective study

Summary Congenital domestic absence of vas deferens (CBAVD) is a common factor in male infertility, and percutaneous epididymal sperm aspiration (PESA) combined with intracytoplasmic sperm injection (ICSI) is a primary clinical treatment, but the effect of the sperm obtained on pregnancy outcome remains to be explored. This study aimed to investigate the relationship between sperm motility with clinical outcome of PESA–ICSI in infertile males with CBAVD. A cohort of 110 couples was enrolled. In total, 76 infertile males were included in the high motility group, while the remaining 34 males were placed in the low motility group. Clinical pregnancy, embryo implantation rate and live birth rate were included as the primary outcome. After all follow-ups, we found that the high motility group achieved higher normal fertilization rates, cleavage rates, transplantable embryo rates and high-quality embryo rates than those in low motility group (normal fertilization rate, 78.2 ± 11.7% vs. 70.5 ± 10.2%, P = 0.003; cleavage rate, 97.1 ± 2.9% vs. 92.3 ± 3.0%, P = 0.000; transplantable embryo rate, 66.8 ± 14.9% vs. 58.6 ± 12.6%, P = 0.009 and high-quality embryo rate, 49.9 ± 10.5% vs. 40.5 ± 11.2%, P = 0.000). Additionally, compared with the low motility group, the clinical pregnancy rates, embryo implantation rates, and live birth rates in the high motility group were significantly increased (pregnancy rate, 61.8% vs. 26.5%, P = 0.009; embryo implantation rate, 36.5% vs. 18.0%, P = 0.044; live birth rate, 55.3% vs. 17.6%, P = 0.000). We concluded that the motility of sperm obtained by PESA affected the clinical outcome of ICSI in infertile males with CBAVD.

Granulocyte Colony-Stimulating Factor Combined With Transcutaneous Electrical Acupoint Stimulation in Treatment of Unresponsive Thin Endometrium in Frozen Embryo Transfer Cycles

Objective: This trial was designed to assess the treatment effects of granulocyte colony-stimulating factor (G-CSF) and transcutaneous electrical acupoint stimulation (TEAS) on thin endometrium in frozen-thawed embryo transfer (FET) cycles.Methods: Ninety-nine patients with previous cancellations of embryo transfer were included, 56 of whom were prospectively treated with intrauterine perfusion of G-CSF in subsequent FET cycles. The selected patients were randomized into the G-CSF perfusion only group and the G-CSF perfusion combined with TEAS group. The other 43 patients were retrospectively included as controls.Results: Compared to previous cycles, endometrial thickness was statistically significantly increased in the two treatment groups (5.97 ± 0.60, 7.52 ± 0.56, 6.14 ± 0.52, and 7.66 ± 0.44; P = 0.00 and 0.00, respectively). The increases in endometrial thickness suggested that no statistically significant difference was found between the two treatment groups. The G-CSF with TEAS group suggested a higher embryo implantation rate than the G-CSF perfusion only and control groups (33.33 and 29.1% and 33.33 and 17.39%; P = 0.412 and 0.091, respectively). The G-CSF combined with TEAS group demonstrated nominally higher clinical and ongoing pregnancy rates than the G-CSF perfusion-only group and controls, though, the difference was not statistically significant.Conclusion: G-CSF has a potential role in improving endometrium thickness in patients with thin unresponsive endometrium in FET treatment cycles. In addition, when combined with TEAS, G-CSF perfusion treatment also improves the embryo implantation rate; however, randomized controlled trials are highly demanded to provide high-grade evidence regarding clinical pregnancy rate after G-CSF perfusion treatment.

P–329 Müllerian anomalies and embryo implantation in oocyte donation

Study question Do patients with Mullerian anomalies (MA) who receive donated oocytes have different embryo implantation rate than patients with normal uterus? Summary answer In oocyte donation, patients with MA had lower implantation rate than patients with normal uterus. What is known already MA are associated with infertility and miscarriage but the mechanisms to explain this relation are not known. Some studies describe both oocyte and/or uterine factor. All studies describing the outcome in patients with MA, so far, are with own oocytes but none in oocyte donation. Study design, size, duration A multicentre restrospective cohort study from January 2000 to December 2019. Patients receiving donated oocytes were divided between those with MA (n = 473) according ESHRE classification and other group with normal uterus (n = 57 869). The primary outcome was implantation rate at fresh embryo transfer. Secondary aims were biochemical pregnancy rate, clinical pregnancy rate, ongoing pregnancy rate, miscarriage rate and live pregnancy rate. Participants/materials, setting, methods We considered the first oocyte donation cycle, without severe male factor, myomas, hydrosalpinx, Asherman syndrome, polyps or indication for preimplantational genetic diagnosis divided in two groups; patients with MA and no malformed uterus. MA group includes cycles of complete bicorporeal uterus (162), partial bicorporeal (30), bicorporeal septate (15), T shaped uterus (26), infantilis uterus (8), complete septate uterus (110), partial septate uterus (94) and hemi-uterus without rudimentary cavity (29). Main results and the role of chance We registered 58 342 patients from our oocyte donation program. Results are shown as mean and 95%CI and differences in pregnancy rates were expressed as relative risks (RR) with 95% CI being reference patients with normal uterus. In patients with MA, the implantation rate was different according the categories being significantly lower in patients with unicornuate uterus (0.29 95%CI: 0.14–0.43. p = 0.03). Biochemical pregnancy rate was significantly higher in patients with septate uterus (RR 1.51 (95%CI 1.02–2.22, p = 0.03) and significantly lower in unicornuate uterus (RR 0.49 (95%CI 0.27–0.90). No differences were found in clinical pregnancy rate among groups, but ongoing pregnancy rate and live birth rate were lower in unicornuate uterus ( RR 0.28 (95%CI 0.13–0.63, p = 0.002), (RR 0.32 (95%CI 0.14–0.73, p = 0.007) respectively. Miscarriage rate was significantly higher in patients with septate uterus (RR 1.78 (95%CI 1.18–2.68, p = 0.006) Limitations, reasons for caution As this was a retrospective cohort study, we were unable to study differences due to modifications in medical or laboratory protocols during this long period time. Different size of sample in some groups of MA makes impossible to translate conclusions to general population. Wider implications of the findings: Our results indicate that there might be a defect in the embryo implantation rate in patients with MA depending on uterine factor. Different sample size among groups and some groups with scarce number of cases make less precise results. More studies controlling biases are needed to confirm our results. Trial registration number NCT04571671

P–778 Repeated cryopreservation process impairs embryo implantation potential but does not affect neonatal outcomes

Study question Does repeated cryopreservation process effect embryo implantation potential and neonatal outcomes of human embryos? Summary answer Repeated cryopreservation impaired embryo implantation potential, resulting in a lower live birth rate and higher miscarriage rate, despite a comparable neonatal complication rate. What is known already With significant advances in the field of ART, the number of available embryos for transfer per cycle has also increased, resulting in a slew of surplus embryo cryopreservation. However, limited researches have focused on the embryonic development potential, clinical outcomes, pregnancy complications as well as the neonatal complications of embryos experiencing repeated cryopreservation. Study design, size, duration This was a retrospective, single-center cohort study. All ART cycles from January 2014 to December 2018. Age, body mass index, and number of oocytes retrieved were preferentially matched within a required range, with a total of 709 couples included in the study. Participants/materials, setting, methods The study was conducted in the Reproductive Medicine Centre affiliated to a university. Preferentially matched participants were divided into three groups according to the times of embryo cryopreservation: the fresh group (n = 249), the cryopreservation group (n = 244) and the re-cryopreservation group (n = 216). Embryo implantation rate, live birth rate, miscarriage rate, and neonatal complication rate were compared among these three groups. Main results and the role of chance The embryo implantation rate, clinical pregnancy rate and live birth rate in the re-cryopreservation group were significantly lower, and there was also a slight increase in the miscarriage rate. Logistic regression analysis indicated that embryos with repeated cryopreservation and lower TE scores were at higher risk of embryo implantation failure in single embryo transfer cycles (OR = 1.79 and 1.56 respectively). No significant differences were observed in gender, gestational age, birthweight, neonatal abnormality, and neonatal complications among the groups. Limitations, reasons for caution This was a retrospective cohort study conducted in single center. A multi-center prospective study with a larger sample size in well-matched participants is needed to reinforce our findings. Wider implications of the findings: Our findings demonstrated the adverse effect of repeated cryopreservation on embryo implantation potential. To avoid embryo waste, or in some special circumstances such as re-biopsy in PGT cycles, an additional cryopreservation on embryos was considered to be available to achieve clinical pregnancy and live birth. Trial registration number Not applicable

P–315 Ultrasound diagnosis of adenomyosis: impact on pregnancy rate in ivf cycles with donated oocytes

Study question The aim was to evaluate in patients who underwent embryo transfer (ET) in an oocyte donation cycle, the impact of adenomyosis, diagnosed by transvaginal sonographic (TVS), on the implantation rate. Summary answer We observed a slightly higher miscarriage rate in the first trimester in patients with adenomyosis in particular in the diffuse type. What is known already What we know from literature is that there are pro studies such as Costello and Vercellini’s which show a reduced pregnancy rate and birth rate, and cons studies which find no effects at all of adenomyosis on IVF treatments. However, both show an increased risk of miscarriage and obstetric complications Study design, size, duration This prospective observational study involved a total of 72 patients: 33 with adenomyosis and 39 without adenomyosis from June 2019 to December 2020. All had a workup which included history, pelvic exam and 2/3D TVS scan which was saved as images, videoclips and volumes and stored. The off line evaluation was performed blind to IVF indication and outcomes by expert sonographer, who assessed the presence or absence of TVS signs of adenomyosis. Participants/materials, setting, methods All the patients aged ≤ 45 years old undergoing, for several personal problems, their first oocyte donation at IVI center Rome.Patients were divided into 2 groups according to findings on a baseline pre-treatment TVS: patients with and without adenomyosis. In the patients with adenomyosis, the disease was further classified according to type (diffuse,focal), localization (inner and outer myometrium) and extension inside the uterus (mild, moderate, severe) and correlated to pregnancy rate and outcome Main results and the role of chance A total of 72 patients were included in this study: 33 with adenomyosis and 39 without adenomyosis. The presence, type and degree of adenomyosis doesn’t show a correlation to embryo implantation rate (64.1% in the control group vs 63.6% in adenomyosis group). However we found an increased risk of early miscarriage in the patients with adenomyosis ( 12% in the control group vs 23.8% in adenomyosis group). Women with adenomyosis that infiltrated only the external myometrium showed a lower pregnancy rate (40%) compared to those who had the involvement of only the inner myometrium (77,7%). The presence of ultrasound findings of focal disease was associated with a lower pregnancy rate (53,3%) compared to the diffuse disease (72,2%); We observed a slightly higher miscarriage rate in the first trimester in patients with adenomyosis in particular in the diffuse type .The presence, type and degree of adenomyosis doesn’t show a correlation to embryo implantation rate. Limitations, reasons for caution Most of the patients included in our study has an age > 40. This could determine an increased number of high-risk pregnancies. Wider implications of the findings: Results of this study may be used to evaluate the impact of different medical or surgical treatment in women with adenomyosis undergoing IVF. Trial registration number Not applicable

Proteomic Analysis of Uterine Tissues During Peri-Implantation Period in Mice with Experimentally Induced Adenomyosis that Treated with anti-Ngf: Implications for Cell-Cell Adhesion and Metabolic Processes

Nerve growth factor (NGF) has been verified to be expressed with higher level in adenomyosis uteri, and its neutralizing antibody has a strong inhibitory influence on inflammation. The present study aimed to explore the effect of anti-NGF on the expression of proteins in uteri of mice-induced adenomyosis and assessed its potential role in improving pregnancy rate. In this study, we established a mouse model of adenomyosis and administrated NGF-neutralizing antibody into mice. The mass spectrometry (MS) analysis of the uteri during the implantation window was performed to explore the essential proteins participating in therapy. Besides, embryos of healthy mice were transferred into the uteri to assess the implantation rate. The results of MS analysis demonstrated that 119 proteins were changed in the adenomyosis group compared with control group, and 126 proteins were differentially expressed in the anti-NGF group compared with the adenomyosis group (fold change > 1.5, P < 0.05. After performing cluster analysis using Mfuzz package, we found that a group of proteins participated in cell-cell adhesion and metabolic processes, which were attenuated in the adenomyosis group, while those were successfully recovered by anti-NGF treatment. Western blotting confirmed that the expression levels of integrin alpha-1 (ITGA1), integrin beta-1 (ITGB1), laminin subunit gamma-1 (LAMC1), and creatine kinase M-type (CKM) were decreased in adenomyosis group, whereas those levels were elevated after anti-NGF treatment. Embryo implantation rate in the adenomyosis group was significantly decreased compared with that in the control group (2.31% vs. 26.15%, P < 0.001) and anti-NGF treatment slightly enhanced the embryo implantation rate in mice with experimentally induced adenomyosis (9.23% vs. 2.31%, P = 0.017). Our results revealed that anti-NGF therapy can improve fertility of mice with experimentally induced adenomyosis, possibly through promoting integrin-related proteins.

Exosomes and soluble secretome from hormone-treated endometrial epithelial cells direct embryo implantation

A successful pregnancy requires a synchronous dialogue between endometrium and embryo within the endometrial milieu. The aim of this study was to assess the role in the implantation of mediators in the endometrial milieu. Total secretome (TS), soluble secretome (SS) and small extracellular vesicles (containing exosomes) were generated from hormonally primed human endometrial epithelial cell culture medium. Human trophectoderm stem cell-derived spheroids were cultured with TS, SS or exosomes (30 µg/ml) on hormonally primed epithelial cells, with exosomes significantly increasing cell adhesion and outgrowth. Furthermore, F1 mouse 2-cell embryos were cultured in groups for 48 h followed by culture with each secretome fraction (30 µg/ml) for 48 h. Blastocyst cell number and hatching were quantified. In addition, blastocysts were further cultured on a fibronectin matrix for 72 h or transferred to recipient mice (with corresponding secretomes) with embryo implantation assessed after 6 days. Exosomes significantly increased total cell number in mouse embryos and complete hatching from zona pellucida, with both exosomes and SS significantly enhancing mouse embryo outgrowth. Importantly, exosomes increased the embryo implantation rate in comparison to other secretome fractions (normalized based on treatment amount) from the endometrial epithelia. These data indicate that endometrial epithelial exosomes support embryo growth, development and implantation while the SS has selective involvement specifically on mouse embryo outgrowth. This finding provides new insights into the molecular differences of endometrial secretome components in implantation and early embryo development and may implicate endometrial exosomes in the pathophysiology of implantation failure in infertility.

Anti-fertility potential of Trigonella foenum-graecum in comparison to oral contraceptive pills

Introduction: This study examines the anti-fertility effects of Trigonella foenum-graecum (fenugreek) seeds extract in the reproductive system of female rats in comparison to combined oral contraceptive pills (COCPs). Methods: Thirty two female Sprague Dawley rats of 8 weeks old were divided into four groups (A, B, C and D). All female rats were allowed to mate with male rats. Group A is a positive control and not given any treatment. Group B is a negative control group. Group C and D were treated with 0.05 mg/kg body weight of COCPs and 750 mg/kg body weight of fenugreek seed aqueous (FSA) extract for 15 days respectively. Blood samples were subsequently taken on the proestrous phase to evaluate the serum levels of follicular stimulating hormone (FSH). The embryo implantation rate was studied in all groups. Results: Administration of 750 mg/kg FSA extract and COCPs both resulted in decreased FSH serum levels, to 25.62 ng/ml and 71.56 ng/ml respectively (p<0.01). The embryo implantation rate was zero in both FSA extract and COCPs treated groups. Conclusions: Anti-fertility effects of FSA extract and COCPs are potentially similar in terms of their ability to reduce FSH serum levels and inhibit implantation.