Severe Male Factor Infertility Research Articles

Effect of intraovarian platelet-rich plasma injection on IVF outcomes in women with poor ovarian response: the PROVA randomized controlled trial.

Does intraovarian platelet-rich plasma (PRP) injection increase the number of mature oocytes obtained after controlled ovarian stimulation (COS) in young women with poor ovarian response (POR) undergoing IVF? Intraovarian PRP injection procedure does not improve mature oocyte yield after COS in women less than 38 years old with an established IVF history of POR. POR is frequently encountered among the infertile population and the number of women seeking infertility treatment related to POR is increasing. Effective treatment options for this patient population to conceive with autologous oocytes are lacking. Case series and cohort studies suggest that intraovarian PRP injection may improve follicular recruitment in women with premature ovarian insufficiency (POI) and POR, yet robust randomized studies have not been performed to date to determine the clinical utility of this intervention. This was a multi-center randomized controlled trial (RCT) conducted at university-affiliated reproductive centers in the USA and Turkey, between January 2020 and November 2022. Patients who met inclusion criteria (<38 years old, two or more prior cycles with <3 oocytes retrieved; and without single gene disorders, prior ovarian surgery, endometriomas, BMI >35 kg/m2, or severe male factor infertility) were randomized to either the PRP or control group. Patients in both groups subsequently underwent COS, oocyte retrieval, ICSI, preimplantation genetic testing for aneuploidy (PGT-A), and single euploid embryo transfer. Number of metaphase II (MII) oocytes obtained was the primary outcome. Secondary outcomes included ovarian reserve tests (antral follicle count [AFC] and anti-Müllerian hormone [AMH]), blastocyst and euploid blastocyst yields, and sustained implantation. The study was powered to detect a difference of one mature oocyte obtained at oocyte retrieval. In total, 83 patients met inclusion criteria and were randomized to receive autologous intraovarian PRP injection (n = 41) or to no intervention (n = 42). No significant differences were observed in number of MII oocytes retrieved per cycle (2.8 ± 2.4 vs 3.1 ± 3.3 in PRP vs control, respectively; P = 0.9), blastocysts (1.0 ± 1.3 vs 1.3 ± 2.1, P = 0.8), or euploid blastocysts (0.8 ± 1.1 vs 0.9 ± 1.6; P = 0.5). Similarly, no differences were observed in the likelihood of obtaining at least one euploid blastocyst (45% vs 37%, P = 0.4; relative risk [RR], 95% CI = 0.9, 0.6-1.2) or the rate of sustained implantation (31% vs 29%, P = 0.9; RR 1.0, 0.7-1.3). Posttreatment AFC (7.9 ± 4.5 vs 6.8 ± 4.8, P = 0.3) and AMH (0.99 ± 0.98 vs 0.7 ± 0.6, P = 0.2) were also not different between the groups. Results from this RCT may not be generalizable to other PRP preparations owing to heterogeneity and lack of standardization. The control groups did not undergo a sham ovarian injection, which would have been relevant had the results shown benefit of PRP injection. Only patients with POR were included in this study, and these results may not be generalizable to more severe diminution of ovarian reserve, as seen with POI. The intraovarian PRP injection procedure does not improve mature oocyte yield or other parameters of IVF outcome in women less than 38 years old with an established IVF history of POR. The results from this study do not support the use of intraovarian PRP injection in this population. Departmental funds were used and no external funding was requested for this study. ES is a consultant for and receives grant funding from the Foundation for Embryonic Competence. All other authors have no conflict of interest to declare. Clinicaltrials.gov Registry Identifier: NCT04163640. 15 November 2019. 24 February 2020.

Outcomes of female reproductive performance with assisted reproductive techniques after recent mild to moderate COVID-19 infections: An observational study.

The purpose of this study was to evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on clinical outcomes among high-risk patients. This retrospective study involved 1,368 patients and the same number of cycles, including 520 cycles with PGT-A and 848 cycles without PGT-A. The study participants comprised women of advanced maternal age (AMA) and those affected by recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), or severe male factor infertility (SMF). PGT-A was associated with significant improvements in the implantation rate (IR) and the ongoing pregnancy rate/live birth rate (OPR/LBR) per embryo transfer cycle in the AMA (39.3% vs. 16.2% [p<0.001] and 42.0% vs. 21.8% [p<0.001], respectively), RIF (41.7% vs. 22.0% [p<0.001] and 47.0% vs. 28.6% [p<0.001], respectively), and RPL (45.6% vs. 19.5% [p<0.001] and 49.1% vs. 24.2% [p<0.001], respectively) groups, as well as the IR in the SMF group (43.3% vs. 26.5%, p=0.011). Additionally, PGT-A was associated with lower overall incidence rates of pregnancy loss in the AMA (16.7% vs. 34.3%, p=0.001) and RPL (16.7% vs. 50.0%, p<0.001) groups. However, the OPR/LBR per total cycle across all PGT-A groups did not significantly exceed that for the control groups. PGT-A demonstrated beneficial effects in high-risk patients. However, our findings indicate that these benefits are more pronounced in carefully selected candidates than in the entire high-risk patient population.

Microfluidic sperm sorting selects a subpopulation of high-quality sperm with a higher potential for fertilization.

Is a microfluidic sperm sorter (MSS) able to select higher quality sperm compared to conventional methods? The MSS selects sperm with improved parameters, lower DNA fragmentation, and higher fertilizing potential. To date, the few studies that have compared microfluidics sperm selection with conventional methods have used heterogeneous study population and have lacked molecular investigations. The efficiency of a newly designed MSS in isolating high-quality sperm was compared to the density-gradient centrifugation (DGC) and swim-up (SU) methods, using 100 semen samples in two groups, during 2023-2024. Semen specimens from 50 normozoospermic and 50 non-normozoospermic men were sorted using MSS, DGC, and SU methods to compare parameters related to the quality and fertilizing potential of sperm. The fertilizing potential of sperm was determined by measurement of phospholipase C zeta (PLCζ) and post-acrosomal sheath WW domain-binding protein (PAWP) expression using flow cytometry, and the chromatin dispersion test was used to assess sperm DNA damage. In both normozoospermic and non-normozoospermic groups, the MSS-selected sperm with the highest progressive motility, PLCζ positive expression and PLCζ and PAWP fluorescence intensity the lowest non-progressive motility, and minimal DNA fragmentation, compared to sperm selected by DGC and SU methods (P < 0.05). The major limitations of our study were the low yield of sperm in the MSS chips and intentional exclusion of severe male factor infertility to yield a sufficient sperm count for molecular experiments; thus testing with severe oligozoospermic semen and samples with low count and motility is still required. In addition, due to ethical considerations, at present, it was impossible to use the sperm achieved from MSS in the clinic to assess the fertilization rate and further outcomes. Our research presents new evidence that microfluidic sperm sorting may result in the selection of high-quality sperm from raw semen. This novel technology might be a key to improving clinical outcomes of assisted reproduction in infertile patients. The study is funded by the Iran University of Medical Sciences and no competing interest exists. N/A.

The effect of laser-assisted hatching on vitrified/warmed blastocysts: the ALADDIN randomized controlled trial

ObjectiveTo evaluate whether laser-mediated assisted hatching (AH) performed on vitrified/warmed blastocysts before embryo transfer can improve live birth rate. DesignThe “pArtiaL zonA pelluciDa removal by assisteD hatchINg of blastocysts (ALADDIN)” is a 2-center comparative study with a parallel randomized controlled design. SubjectsParticipants were recruited between September 2018 and November 2021. They were aged 18 to 39 years, underwent non-donor IVF cycles, and were scheduled for elective single embryo transfer with vitrified/warmed blastocysts. Those with uterine abnormalities, BMI >35 kg/m2, severe male factor infertility, or performing preimplantation genetic testing were excluded. InterventionAssisted hatching was performed using a 1480 nm diode laser, removing approximately one-third of the zona pellucida with continuous 0.2 ms pulses applied from the 1 to 5 o’clock positions. Main Outcome MeasuresThe primary outcome was the live birth rate. Secondary endpoints included clinical pregnancy, miscarriage, multiple pregnancies, preterm births, obstetrical and neonatal complications, and congenital anomalies. ResultsOverall, 698 participants met the inclusion criteria and were randomized: 352 patients were assigned to the AH arm and 346 to the control arm. One-hundred and five (29.8%) and 101 (29.2%) participants achieved a live birth following treatment, respectively (p=0.87). The relative risk of live birth in patients with vitrified/warmed blastocysts treated with AH was 1.02 (95%CI: 0.86-1.19). Exploratory subgroup analyses for women’s age, recruiting centers, indications for IVF, method of insemination, blastocyst quality, and days of blastocyst development failed to highlight any clinical situation that could benefit from AH in thawed blastocysts. ConclusionIn patients undergoing frozen embryo transfer with vitrified/warmed blastocysts, laser AH does not improve the live birth rate. Further studies are required to rule out milder but potentially interesting benefits in specific subgroups of patients.

Embryological and clinical outcomes in couples with severe male factor infertility versus normozoospermia

ObjectiveThis study evaluated embryological and clinical outcomes in couples with severe male factor infertility versus those with normozoospermia undergoing ICSI and in vitro fertilisation. MethodsThis multicentre, retrospective cohort study included all couples who had undergone autologous ICSI cycles at My Duc Hospital and My Duc Phu Nhuan Hospital in Vietnam between January 2018 and January 2021 (female age < 35 years and males with severe male factor or normozoospermia based on the World Health Organization 2010 criteria). The primary outcome was the cumulative live birth rate after the first ICSI cycle. ResultsA total of 1296 couples were included, including 648 with severe male factor infertility and 648 with normozoospermia. The number of two pronuclei zygotes, embryos, and frozen embryos was significantly lower in couples with severe male factor infertility compared with normozoospermia (p < 0.05). In contrast, there were no significant differences between the two groups with respect to cumulative pregnancy outcomes, including the live birth rate, and secondary outcomes including clinical pregnancy rate, ongoing pregnancy rate, and miscarriage rate. ConclusionSevere male factor infertility appeared to have an impact on the fertilisation and early developmental potential of embryos, but sperm quality did not affect cumulative clinical fertility outcomes.

Clinical outcomes of preimplantation genetic testing for aneuploidy in high-risk patients: a retrospective cohort study.

The purpose of this study was to evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on clinical outcomes among high-risk patients. This retrospective study involved 1,368 patients and the same number of cycles, including 520 cycles with PGT-A and 848 cycles without PGT-A. The study participants comprised women of advanced maternal age (AMA) and those affected by recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), or severe male factor infertility (SMF). PGT-A was associated with significant improvements in the implantation rate (IR) and the ongoing pregnancy rate/live birth rate (OPR/LBR) per embryo transfer cycle in the AMA (39.3% vs. 16.2% [p<0.001] and 42.0% vs. 21.8% [p<0.001], respectively), RIF (41.7% vs. 22.0% [p<0.001] and 47.0% vs. 28.6% [p<0.001], respectively), and RPL (45.6% vs. 19.5% [p<0.001] and 49.1% vs. 24.2% [p<0.001], respectively) groups, as well as the IR in the SMF group (43.3% vs. 26.5%, p=0.011). Additionally, PGT-A was associated with lower overall incidence rates of early pregnancy loss in the AMA (16.7% vs. 34.3%, p=0.001) and RPL (16.7% vs. 50.0%, p<0.001) groups. However, the OPR/LBR per total cycle across all PGT-A groups did not significantly exceed that for the non-PGT-A groups. PGT-A demonstrated beneficial effects in high-risk patients. However, our findings indicate that these benefits are more pronounced in carefully selected candidates than in the entire high-risk patient population.

A Comparison of ICSI Outcomes and Reactive Oxygen Species Levels in Seminal Plasma between Normozoospermia and Sperm Abnormalities Groups for Infertile Men

Background: Social life and the healthcare systems are affected by infertility. Infertility could be primary or secondary; secondary infertility is the inability to conceive after one successful pregnancy while primary infertility is the inability to conceive at all. As sperm abnormalities cause male infertility, Intracytoplasmic Sperm Injection (ICSI) is a particular type of in vitro fertilization (IVF) used to treat severe male-factor infertility. Free radical oxygen derivatives are reactive oxygen species (ROS). Reactive oxygen species may cause 30% to 80% of male infertility. Aims of the study: the present study aims to determine the reactive oxygen species level in seminal plasma and to study the Intracytoplasmic Sperm Injection outcomes for Normospermia group and compare it with Oligozoospermia, Asthenozoospermia and the Teratozoospermia Groups. Material, and Methods: a cross-sectional study was conducted between January 2023 to June 2023 in Najaf, on a non-random sampling including 50 couples who suffered from a minimum of 12 months of primary fertility with regular unprotected sexual intercourse and who had attended the fertility center in Najaf requesting fertility treatment. A gynecological examination and assessment were done for the female participants while the urologists examined and assessed the male subjects; then, a semen analysis was carried out followed by macroscopic and microscopic examinations. After the preparation of the sample for ICSI and performing ICSI, pregnancy was assessed and the outcomes of the different groups were compared. At the same time, reactive oxygen species levels were assessed by using the Human Reactive Oxygen Species ELISA kit. Then, the SPSS version 26 was used to perform the statistical analysis. Results: Among the twenty-one patients with Normozoospermia, seven with Asthenospermic, seven with Oligozoospermia and fifteen with Teratozoospermia, there was a significant difference among the four groups regarding sperm concentration, progressive motility, and sperm normal morphology. There was no significant difference among the type of sperm, Normo, Astheno, Tearato, and Oligozoospermia, regarding birth rate and ICSI outcomes. Males with higher progressive motility showed a higher pregnancy rate. In conclusion, despite the presence of reactive oxygen species, Intracytoplasmic Sperm Injection was advantageous and effective for all four groups. reactive oxygen species had no significant effect on Intracytoplasmic Sperm Injection outcomes, however Normozoospermia was associated with greater pregnancy rates. Despite the presence of reactive oxygen species, Intracytoplasmic Sperm Injection was advantageous and effective for all four groups. ROS had no significant effect on Intracytoplasmic Sperm Injection outcomes, however Normozoospermia was associated with greater pregnancy rates.

Levothyroxine for a high-normal TSH in unexplained infertility.

Unexplained infertility affects nearly one-third of infertile couples. Women with unexplained infertility are more likely to have a high-normal thyroid-stimulating hormone level(TSH: 2.5-5 mIU/L) compared to women with severe male factor infertility. Practice guidelines vary on whether treatment should be initiated for TSH levels >2.5 mIU/L in women attempting conception because the effects of treating a high-normal TSH level with levothyroxine are not known. We evaluated conception and live birth rates in women with unexplained infertility and high-normal TSH levels. Retrospective study including 96women evaluated for unexplained infertility at a large academic medical centre between 1 January 2000 and 30 June2017 with high-normal TSH (TSH: 2.5-5 mIU/L and within the normal range of the assay) who were prescribed (n = 31) or not prescribed (n = 65) levothyroxine. Conception and live birth rates were assessed. The conception rate in the levothyroxine group was 100% compared to 90% in the untreated group (p = .086 unadjusted; p < .05 adjusted for age; p = .370 adjusted for TSH; p = .287 adjusted for age and TSH).The live birth rate was lower in the levothyroxine group (63%) compared to the untreated group (84%) (p = .05 unadjusted; p = .094 adjusted for age; p = .035 adjusted for TSH; p = .057 adjusted for age and TSH). Women with unexplained infertility and high-normal TSH levels treated with levothyroxine had a higher rate of conception but lower live birth rate compared to untreated women, with the limitation of a small sample size. These findings assert the need for prospective, randomized studies to determine whether treatment with levothyroxine in women with unexplained infertility and high-normal TSH is beneficial.

#249 : Spotlight on the Artefacts in Next Generation Sequencing in PGT-A: Possible Reason for High Mosaicism Reporting

Background and Aims: Advancement in Next Generation Sequencing (NGS) has led to the extensive utilization of Preimplantation Genetic testing for Aneuploidy. But one challenge, in reporting of aligned short read sequencing data, is the ability to discriminate between embryonic NGS profiles with genuine pathological copy number variations (CNVs) from artefacts arising from erroneous DNA amplification/library preparation. Do technical errors in VeriSeq-NGS lead to artefacts in CNV charts resulting in falsely increased reporting of chromosomal mosaicism? Method: After ethical clearance and informed consent, patients [Formula: see text] 35 years, [Formula: see text]1 implantation failures and severe male factor infertility underwent PGT-A. Those with aneuploid embryos on TE biopsy, donated their embryos for research in year 2022. The DNA extraction and amplification was carried out using Sureplex-DNA amplification system. The libraries were prepared using VeriSeqTM-PGS Library Preparation kit. A total of 24 libraries were pooled, denatured and subjected to NGS using Illumina MiSeq system. CNV visualization and analysis for each sample was carried out using BlueFuse Multi Software (Illumina, USA). The aneuploid whole blastocyst were subjected to the same protocol. Results: NGS profiles generated from the aneuploid whole blastocysts, were manually analyzed for CNVs and for any common genomic artifacts. The quality control was performed using Illumina-Quality Control requirements and only samples satisfying these requirements were used for analysis in the study. Common artifacts were identified affecting chromosome 7,11,16 and 19. Out of 22 blastocysts, NGS profiles of 21 blastocysts had artifact at chromosome 19, and 15 had artifacts at chromosome 7,11,16. These artifacts may be introduced during whole genome amplification and/or suboptimal NGS library preparation or is inherent weakness of the VeriSeq-NGS kit. Conclusion: The four commonly occurring artefacts involved chromosome 7,11,16 and 19, which could be introduced during steps of whole genome amplification or library preparation. The awareness of these artefacts is important to reduce the false positive/inconclusive rate.

ICSI outcome after microdissection testicular sperm extraction, testicular sperm aspiration and ejaculated sperm

We conducted a case-controlled single-center cohort study to evaluate the intracytoplasmic sperm injection (ICSI) outcome in severe male infertility with different methods of sperm obtention. The data was compiled from a tertiary university hospital. The micro-TESE procedures were performed from 2008 to 2023, with a sperm recovery rate (SRR) of 45 %. The ICSI treatments were carried out between 2011 and 2023. The aim of the study was to compare the ICSI outcome using sperm obtained by microdissection testicular extraction (micro-TESE), testicular sperm aspiration (TESA), and ejaculated sperm with sperm concentration less than 15 million per milliliter. We included a total of 462 ICSI cycles, of which 340 ICSIs with ejaculated sperm of men with oligozoospermia, with or without asthenozoospermia or teratozoospermia (OAT group), 51 ICSIs with TESA sperm of men with obstructive azoospermia (OA, TESA group), and 71 ICSIs with micro-TESE sperm of men with non-obstructive azoospermia (NOA, micro-TESE group). The patient characteristics, fertilization rate, pregnancy rate, and pregnancy outcome data were similar between the groups. The fertilization rates were 66.0 % in the OAT group, 68.3 % in the TESA group and 62.8 % in the micro-TESE group and live birth rate per embryo transfer were 23.7 %, 28.9 %, and 25.0 %, respectively, without statistical difference. The obstetrical outcome was similar in all the groups. The overall clinical results in all ICSI cycles performed for treating severe male factor infertility were similar, independent of the method of collection of spermatozoa. The results also confirm the efficacy of micro-TESE in the treatment of severe male factor infertility.

Intracytoplasmic sperm injection does not improve the outcome of IVF treatments in patients with advanced maternal age or low oocyte number: A randomized controlled trial.

Intracytoplasmic sperm injection (ICSI) was introduced to achieve fertilization in cases of severe male factor infertility. However, ICSI is often used in cases of non-male factor infertility, such as advanced maternal age or low oocyte number, but the clinical benefit of the method in these indications has not been proven. A prospective randomized study was conducted in a university clinic between 2018 and 2020. Patients with ≥40 years of age and/or ≤4 oocytes with non-sever male factor infertility were randomized into conventional IVF or ICSI groups. Fertilization rate, embryo quality, implantation, clinical pregnancy and live birth rates were compared. A total of 336 IVF cycles (169 conventional IVF and 167 ICSI) were involved in the study. The fertilization rate was higher in the conventional IVF group compared to the ICSI group (IVF: 61.7%, ICSI: 53.4%, P=0.001). Embryo development and morphology did not show considerable difference between groups. Implantation, clinical pregnancy and live birth rate were 13.1%, 24.3% and 11.4% in the conventional IVF and 10.4%, 19.0%, 12.0% in the ICSI group. The differences were not significant. Subgroup analysis showed a significantly better clinical outcome following conventional IVF when advanced maternal age was accompanied by low oocyte number (Implantation: 11.7% vs 2.6%, P=0.027; Clinical pregnancy: 18.5% vs 4%, P=0.020). A significantly higher fertilization rate, a tendency for higher clinical pregnancy rate was found in conventional IVF treatments compared to ICSI. When advanced maternal age was associated with low oocyte number, ICSI resulted in a substantially lower chance of fertilization and clinical pregnancy. These data suggest that ICSI offers no advantage over conventional IVF in terms of fertilization, embryo quality, implantation and pregnancy rates for couples with advanced maternal age or with low oocyte number.

Manipulated sperm motility via soot nanoparticles-induced biochemical alterations in human seminal plasma

Obtaining spermatozoa with progressive motility, via postejaculatory activation with pharmacological agents such as theophylline and pentoxifylline, is crucial for the success rate of assisted reproduction in couples with severe male factor infertility. Regrettably, the possibility of premature acrosome reactions and impared oocyte function questions the practical applicability of phosphodiesterase inhibitors. The rapid development of nanotechnologies promotes the use of hydrophobic rapeseed oil soot as a non-cytotoxic biomaterial for sperm motility activation, but the scarcity of knowledge regarding the interactions of soot with components from the seminal plasma hinders the eventual commercialization of this cutting-edge approach. Aiming to eliminate this shortcoming, the current study shows for the first time how the soot nanomaterials alter the biochemistry of human seminal plasma. Upon 270 min incubation with soot nanoparticles, the activity of AST, ALT, CK, LDH and GGT enzymes in the seminal plasma of ten patients changes inversely to the registered sperm motility (i.e., lower enzyme activity, higher sperm motility and vice versa). This phenomenon is primarily related to termination of the enzymes-substrate binding or extraction of enzymes from the gametes via chemical bonding with the soot. These novel mechanisms depend on the physicochemical features of used carbon nanomaterials, revealing opportunities for predictable tuning of the sperm reproductive potential.

O-065 A multi-center randomized controlled trial of intraovarian injection of platelet rich plasma for women with poor ovarian response

Abstract Study question Does intraovarian injection of autologous platelet-rich plasma (PRP) improve oocyte yield in young patients with poor ovarian response (POR)? Summary answer Intra-ovarian PRP injection did not result in increased number of oocytes retrieved or improvement of any other relevant measured outcomes. What is known already POR is one of the major hurdles to overcome in patients undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). Several methods have recently emerged to attempt follicular reactivation in these patients with a poor prognosis. One of them, the intra-ovarian injection of autologous PRP, demonstrated promising results in retrospective and prospective cohort studies. Study design, size, duration This was a multi-center, randomized controlled trial (RCT) to evaluate the efficacy of PRP in improving IVF outcomes in women with POR. Patients who met inclusion criteria (&amp;lt;38 years, two or more prior cycles with &amp;lt; 3 oocytes retrieved, and without single gene disorders, prior ovarian surgery, endometriomas, BMI &amp;gt;35, or severe male factor infertility) were randomized to either receive an autologous intra-ovarian PRP injection (PRP group) or no intervention (control group) prior to COH. Participants/materials, setting, methods A total of 83 patients were randomized to PRP group (n = 41) or control group (n = 42). After treatment, patients underwent COH, oocyte retrieval, intracytoplasmic sperm injection (ICSI), preimplantation genetic testing for aneuploidy (PGT-A), and single frozen euploid embryo transfer. Number of MII oocytes obtained was the primary outcome. Secondary outcomes included ovarian reserve tests (antral follicle count [AFC] and anti-Mullerian hormone [AMH]), blastocyst and euploid blastocyst yields, and sustained implantation. Main results and the role of chance No significant differences were observed in number of MII oocytes retrieved (3.1 ± 3.3 vs 2.8 ± 2.4 in PRP vs control, respectively; p = 0.9), blastocysts (1.3 ± 2.1 vs 1.0 ± 1.3, p = 0.8), or euploid blastocysts (0.9 ± 1.6 vs 0.8 ± 1.1; p = 0.5) per cycle. Similarly, no differences were observed in the likelihood of obtaining at least one euploid blastocyst (37 vs 45%, p = 0.4; relative risk [RR], 95% confidence interval [CI] 0.9, 0.6-1.2) or the rate of sustained implantation (29 vs 31%, p = 0.9; RR 1.0, 0.7-1.3). Pre- and post-treatment AFC in both groups (PRP: from 5.2 ± 3.2 to 7.9 ± 4.5, p &amp;lt; 0.0001; control: from 5.6 ± 3.3 to 6.8 ± 4.8, p = 0.02) were statistically significantly different, whereas AMH was different in the PRP group only (PRP: 0.73 ± 0.46 to 0.99 ± 0.98, p = 0.02; control: 0.70 ± 0.59 to 0.73 ± 0.57, p = 0.59). Limitations, reasons for caution Oocyte retrieval took place the cycle immediately following treatment; therefore, we could not evaluate long-term effects of PRP on ovarian reserve or response. Lack of a placebo group would limit interpretability had a difference between groups been observed. The effect of PRP on patients &amp;gt;38 years old was not studied. Wider implications of the findings Intra-ovarian PRP injection did not improve reproductive outcomes by any relevant measure. Our findings do not support wide utilization of PRP to improve IVF outcomes in women with POR. Trial registration number NCT04163640

P-315 To study the efficacy of Transvaginal ultrasound-guided Embryo Transfer over Transabdominal ultrasound-guided Embryo Transfer in transfers with poor visibility due to obesity or marked retroversion

Abstract Study question Why is Transvaginal guided embryo transfer,understudied and underutilised in difficult cases of embryo transfers with compromised visibility of uterine canal in Transabdominal ultrasound ? Summary answer In patients with poor visibility of uterine canal using TAUS due Obesity, retroveted uterus,PID,pelvic adhesions, endometriosis, TVUS-ET gives significantly higher clinical Pregnancy rates. What is known already TAUS-ET is most popular method of Embryo transfer worldwide. Despite the benefits , this procedure also presents some important drawbacks. Firstly, TAUS-guided ET requires an assistant to operate the transducer while physician performs ET procedure. Moreover, patient needs to fill her urinary bladder to enable visualization of uterine canal, which causes discomfort and sometimes uterine cramping during ET, which in turn can negatively impact implantation rates.TVUS is the main technique used in all other gynecological investigations. Because of high frequency and close proximity to target area, TVUS transducers provide better resolution of the uterocervical angle and improved overall image quality. Study design, size, duration This retrospective study was conducted to compare efficacy of TVUS-guided ET to conventional TAUS-guided ET in difficult transfers, who underwent treatment at Triveni IVF, Darbhanga, India between November 2020 to December 2022. It was based on data extracted from records of all women who underwent difficult TAUS-ET procedure between Nov 2020 to Nov 2021.Data from records of matching number of women who underwent TVUS-ET procedure between Dec 2021 to Dec 2022 were used for comparison. Participants/materials, setting, methods All were ICSI and FET transfers. Inclusion criteria: (1) All included between 25–35 years of age with difficult visualisation of uterine canal during TAUS-Mock ET in previous cycle (2) BMI ≥ 30; (3) Retroverted uterus due to any underlying pathology 4)All pts included had good ovarian reserve; (4) unexplained infertility &amp;gt;3 years. Exclusion criteria: (1) severe male factor infertility, (2) any intrauterine factor of infertility (3) ovarian endometrioma or PCOS and (5) multiple ICSI failure (&amp;gt;3 trials). Main results and the role of chance Total of 123 cycles,61 guided by TVUS (Group 1) and 62 TAUS (Group 2) were included. Our results showed significantly higher Clinical Pregnancy rate in TVUS group as compared to TAUS group, in difficult transfers (43.3% vs. 36.1 % p = 0.0045). VAS (visual analog scale) of the pain during ET was significantly less in TVUS group in comparison to TAUS group (17% vs 64% p = 0.0001), also the abdominal discomfort was significantly less in TVUS group in comparison to TAUS group (11% vs. 69 % p = 0.0001). TVUS-ET group showed significantly higher CPR as it has better resolution due to proximity to target organs allowing proper visualization of catheter tip especially in cases with obesity and marked uterine retroversion.TVUS has superior role in cases with difficult transfer because it can easily analyze the cause of difficult transfers as endocervical crypts, marked anteverted uterus, cervical canal tortuosity. Another advantage of TVUS is requiring emptied urinary bladder in contrast to TAUS. Full bladder is both time consuming and causes abdominal discomfort and uterine cramps leading to increased patient anxiety.As regards to abdominal discomfort and uterine cramps, there was extremely high statistical significance between the two groups in our study. Limitations, reasons for caution Despite the wide benefits,TVUS-ET has not been widely adopted, on account of difficulty in manipulating ET catheter and vaginal transducer simultaneously in very constricted area.Other comfortable approaches that may assist physicians such as inner catheter can be pushed by embryologist while physician holds TVS-probe and outer catheter in place. Wider implications of the findings The results suggested that TVUS-guided ET was better in difficult transfers with poor visibility of uterine canal in studied sample. This technique presented deserves consideration for wider use since it can offer better visualization, excellent comfort to patients during the procedure, and requires only one operator. Trial registration number Not applicable

P-143 Will Use of Testicular sperm in ICSI cycles increase Embryo Aneuploidy?

Abstract Study question Will use of surgically retrieved Testicular Sperms (TESA) in Intra-Cytoplasmic Sperm Injection (ICSI) cycles increase embryo aneuploidy? Summary answer Use of TESA sperms for ICSI seems safe and doesn’t increase embryo aneuploidy. TESA performed for various indications also showed no variation with embryo aneuploidy. What is known already Invention of ICSI and improved surgical sperm retrieval techniques have helped men with severe male factor infertility to father their own genetic child. Sperms retrieved from seminiferous tubules of testes are considered immature as they haven’t completed the process of spermiogensis. It is hypothesized that use of TESA sperm for ICSI can increase embryo aneuploidy, which is still uncertain. Study design, size, duration This retrospective study conducted at a private fertility clinic from 2014–2022 (n = 265 patients; total of 860 embryos evaluated). Study populationTESA sperm used and PGT-A done (n = 66). TESA group was further sub-divided into 2 groups based on the indication for TESA,Non-Obstructive Azoospermia(n = 26) and Raised SDF(n = 40). Control Population Ejaculated sperm used and PGT-A done (n = 199) Only younger women(&amp;lt;37yrs) with self-gametes and RIFconsidered in this study. RIF was defined as women with 2 failed IVFcycles in thepast with at least 4 blastocystsbeing transferred. Participants/materials, setting, methods All fertilized oocytes inseminated by ICSI and subjected to extended blastocyst culture. TESA done as per our clinic’s SOP and for control group sperm preparation done by density gradient method. Trophectoderm biopsy was performed and sent for PGT-A to the genetic lab. Next-Generation sequencing (NGS) was the Comprehensive Chromosomal technique (CCS) used for assessing embryo ploidy status. Aneuploidy across all groups compared. Main results and the role of chance Following were the Aneuploidy rates of TESA sperm, Ejaculate sperm group and TESA done for NOA and SDF respectively. Ejaculate Sperm – Aneuploidy%- 42% TESA Sperm (irrespective of the indication)- Aneuploidy%- 44% (p = 0.6260) TESA Sperm (done for NOA)- Aneuploidy%- 44% (p = 0.7504) TESA Sperm (done for raised SDF) – Aneuploidy%- 32% (p = 0.05) Embryo aneuploidy was comparable between Ejaculate and TESA sperm group. Though TESA sperm is considered immature, it didn’t seem to increase embryo aneuploidy. Embryo Aneuploidy from TESA sperm ICSI cycles done for NOA or raised SDF also showed no altering trend. Data from this study is suggestive that TESA sperm doesn’t alter embryo ploidy status. Limitations, reasons for caution Data is retrospective with small sample size and unequal distribution. A well designed Randomized Trial might help test the inference of this study further. Wider implications of the findings Incidence of male infertility and use of ICSI is on a rise globally. Interventions to optimize embryo implantation and reproductive outcomes will help with better success rates and shorten duration to conception. Further research is warranted with severe male factor infertility to improve sperm selection techniques and reproductive outcomes. Trial registration number not applicable

O-136 Artificial intelligence to assist in surgical sperm detection and isolation

Abstract Study question Can an artificial intelligence (AI) improve the speed and accuracy of identifying sperm in complex testicular tissue samples? Summary answer Trained AI can identify sperm in real-time instantly with higher accuracy, not only reducing strain on embryologists but increasing sample coverage in a shorter time. What is known already Non-obstructive azoospermia (NOA) is a form of severe male-factor infertility, affecting nearly 5% of infertile couples seeking treatment. Isolating sperm from macerated testicular tissue for intracytoplasmic sperm injection (ICSI) has changed marginally in the last two decades, and still requires embryologists to search arduously through a background of collateral cells including red blood cells (RBC’s), white blood cells (WBC’s), leydig, sertoli and epithelial cells, causing fatigue and reducing sample coverage. Image analysis using trained AI can instantly identify shapes and forms and thus presents itself as a candidate to dramatically reduce processing times in surgical sperm cases. Study design, size, duration This proof-of-concept consists of two phases over 5 months. A training phase using 7 azoospermic patients to provide samples to train a convolutional neural network with manual annotation. Secondly, a side-by-side live test with an embryologist versus the AI model comparing time taken and accuracy of sperm identification, and precision of identifying sperm in macerated tissue samples (false positives and false negatives). Results were analysed using Mann-Whitney U-test. Differences were considered significant when p-value&amp;lt;0.05. Participants/materials, setting, methods Consenting patients at IVF Australia, Sydney attending for surgical sperm collection donated discarded macerated testicular tissue samples whereby excess tissue was used post-processing in conjunction with donor sperm, to train the machine learning software. Thereafter, testing using conventional ICSI micromanipulator microscope was performed side-by-side on test samples and time taken, accuracy and precision were recorded between a control count, computer vision and trained embryologist. Main results and the role of chance The AI model showed dramatic improvement in time taken to identify sperm per field, improved accuracy in identifying sperm as well as a high level of precision. Precision for the trained embryologist was considered to be 100%. Time taken per field of view to identify all sperm was significantly lower for the AI when compared to the trained embryologist (0.019 ± 1.4 x 10-4s vs 22.87 ± 0.98s, P &amp;lt; 0.0001). There was also a significant difference in accuracy (89.88 ± 1.56% vs 83.22 ± 2.02%, P = 0.017) for the AI model versus the trained embryologist respectively. The precision of the model is 91.27 ± 1.27% which considered false positives and the correct identification of sperm versus the control count. From a total of 688 sperm to be found, 560 were found by the embryologist and 611 were found by the AI model in less than 1000th of the time it took the embryologist. Limitations, reasons for caution This is a proof-of-concept test of a convolutional neural network AI. Following this study, side-by-side on donated clinical samples are needed, and a clinical trial comparing embryologists versus AI is required to prove efficacy and usefulness. A larger number of samples across multiple indications and surgical approaches is required. Wider implications of the findings AI-powered image analysis has the potential for seamless integration into laboratory workflows to reduce time to identify and isolate sperm from surgical sperm samples from hours to minutes, and do this with improved accuracy, thus increasing success rates from these treatments. Trial registration number N/A

ICSI for non-male factor infertility: time to reappraise IVF?

Intracytoplasmic sperm injection (ICSI) was initially introduced to overcome problems due of severe male factor infertility not being solved with conventional in-vitro fertilization (cIVF). However, recent years have witnessed an increasing use of ICSI by most assisted reproductive technique laboratories for non-male factor indications. Examples of the latter include previous fertilization failure after cIVF, few or poor-quality oocytes, immature oocytes, advanced maternal age, preimplantation genetics test (PGT), cryopreserved oocytes, and unexplained infertility. The replacement of cIVF with ICSI in several non-male factor infertility cases is probably because some reproductive specialists consider that ICSI is associated with better reproductive outcomes. Unfortunately, data on reproductive outcomes in favor of ICSI over cIVF are limited or absent. Therefore, the factors that can help define the use of one technique over the other should be identified. These should include the likelihood of fertilization failure, potential risks of the procedure, and its costs. In this review, we aim to highlight the current guidelines, advantages, and limitations of the use of cIVF/ICSI for infertility treatment. Additionally, we provide a comprehensive review of the use of ICSI in indications other than severe male factor infertility.

First pilot study of maternal spindle transfer for the treatment of repeated in vitro fertilization failures in couples with idiopathic infertility

To gain insights into the technical feasibility of maternal spindle transfer (MST) applied in the context of repeated invitro fertilization (IVF) failures for the treatment of idiopathic infertility. A prospective pilot study. IVF center. Twenty-five infertile couples with multiple previous unsuccessful IVF cycles (range, 3-11), no previous pregnancy, and no history of mitochondrial DNA (mtDNA) disease participated. The study focused on women <40 years, with previous IVF attempts characterized by a pattern of low fertilization rates and/or impaired embryo development. Couples with severe male-factor infertility were not eligible. Oocyte donors with previous successful IVF outcomes were matched with patients according to standard practice. We performed MST by transferring metaphase II spindles from the patients' oocytes into the previously enucleated donor oocytes, followed by intracytoplasmic sperm injection, invitro embryo culture, blastocyst biopsy, and vitrification. Only euploid blastocysts were considered for embryo transfer. Outcome measures included oocyte fertilization, blastocyst development, clinical pregnancy and live birth, incidence of mitochondrial carryover and potential mtDNA reversal, as well as general health of the children born. Twenty-eight MST cycles produced 6 children (19 embryo transfers, 7 clinical pregnancies). Pediatric follow-up of the children, performed at intervals from birth to 12-24 months of age, revealed their development to be unremarkable. DNA fingerprinting confirmed that the nuclear DNA of MST children was inherited from both parents, without any contribution from the oocyte donor. For 5 of the children, mtDNA was derived almost exclusively (>99%) from the donor. However, 1 child, who had similarly low mtDNA carryover (0.8%) at the blastocyst stage, showed an increase in the maternal mtDNA haplotype, accounting for 30% to 60% of the total at birth. This pilot study provides the first insights into the feasibility of applying MST for patients with idiopathic infertility and repeated IVF failures. Reconstructed oocytes produced embryos capable of implanting, developing to term and producing apparently healthy newborns/children. However, claims concerning the efficacy of MST with respect to infertility treatment would be premature considering the limitations of this study. Importantly, mtDNA reversal was detected in one child born after MST, a finding with possible implications for mitochondrial replacement therapies. Pilot trial registry number, ISRCTN11455145. The date of registration: 20/02/2018. The date of enrolment of the first patients: 18/03/2018.

Outcome analysis of ICSI assisted pregnancy using testicular sperm versus ejaculated sperm in man with severe oligozoospermia in the same ART cycle: A case report.

Intracytoplasmic sperm injection (ICSI) has become the most common method for couples with male factor infertility, and source of sperm for the procedure have evolved over time. but few have examined testicular sperm extraction vs. ejaculated sperm use for severe oligozoospermia in the same assisted reproductive technology (ART) cycle. Here, we evaluated the clinical outcomes after ICSI with testicular sperm or ejaculated in man with severe oligozoospermia in the same ART cycle. A couple who had failed the first ART cycle with ejaculated sperm, using the freshly ejaculated sperm and testicular sperm for ICSI during the second ART cycle by lack of enough sperm to fertilize in an ICSI attempt. The patient was diagnosed with severe oligozoospermia, and routine semen analysis revealed sperm concentration is less than 2 million/mL. The patient using testicular sperm versus ejaculated sperm with ICSI assisted pregnancy in the same ART cycle. We found that superior cleavage rate, number of embryos transferred and blastocyst rate with the use of testicular rather than ejaculated sperm-ICSI in the couple. The results described here suggest that use of testicular sperm may improve biologic outcomes, especially for couples with male-partner oligozoospermia who previous ICSI failures. Our case report supported the efficacy of testicular sperm preference over ejaculated sperm for ICSI in men with severe male factor infertility. It is a paradigm shift concerning the use of ejaculated sperm as the preferable source of sperm for ICSI, add to the small amount of literature on testicular sperm extraction vs. ejaculated sperm use for severe oligozoospermia in the same ART cycle.

The impact of small and asymptomatic intramural and subserosal fibroids on female fertility: a case-control study.

Do small and asymptomatic intramural and subserosal uterine fibroids affect female fertility? Small and asymptomatic fibroids that do not encroach the endometrial cavity appear to not markedly affect female fertility. The available evidence on uterine fibroids and fertility is limited. Most information has been obtained in IVF settings by comparing the success in women affected and not affected by fibroids. These studies have shown a detrimental effect of submucosal and possibly intramural fibroids. However, this study design provides information only on embryo implantation, not on female fertility in general. A retrospective observational case-control study on 200 women whose partner was diagnosed with severe male infertility and 200 women with unexplained infertility was conducted. If the null hypothesis (that fibroids do not affect fertility) is valid, one would expect a similar prevalence of fibroids in the two study groups. Conversely, if fibroids do impact fertility, one would expect a higher prevalence among women with unexplained infertility. The study was carried out at the Infertility Unit of the Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico covering a 5-year period between January 2014 and June 2020. We retrospectively recruited women seeking pregnancy whose partner was repeatedly documented to have a sperm concentration below 1 million/ml and matched them by age and study period to a group of women with unexplained infertility. The latter group of women was considered as a case group (infertile subjects), while the former group of women was considered as a control group (reflecting the general female population). Women with fibroids could be included in both study groups; only those with submucosal lesions were excluded. Fibroids were diagnosed in 31 women (16%) with unexplained infertility and in 32 women (16%) with severe male factor infertility. The adjusted odds ratio of carrying fibroids in women with unexplained infertility was 0.91 (95% CI: 0.52-1.58). Subgroup analyses according to number, dimension and location of fibroids failed to highlight an increased risk of infertility in any group. This is a retrospective study and some inaccuracies in fibroids detection cannot be ruled out. Moreover, the relatively small sample size hampers robust subgroup analyses. Even though we excluded women with patent causes of infertility, some women with specific causes of infertility could have been included among controls (yet are expected to account for <10% of the group). This study suggests that small fibroids that do not encroach the endometrial cavity do not markedly affect female fertility. This information is clinically relevant when counseling infertile women with small fibroids and an otherwise unremarkable diagnostic work-up. Surgery may still be considered but only in selected cases. This study was partially funded by Italian Ministry of Health: current research IRCCS. E.S. reports grants from Ferring, grants and personal fees from Merck, and grants and personal fees from Theramex outside the submitted work. All the other authors do not have any competing interest to declare. N/A.