ICSI Patients Research Articles

P-062 The impact of isolated teratozoospermia on cumulative live birth rates in IVF and ICSI treatment: a retrospective analysis of 5254 completed cycles

Abstract Study question To assess fecundity using cumulative live birth rate (CLBR) per egg collection to determine if IVF and ICSI outcomes are significantly influenced by sperm morphology. Summary answer The presence of isolated teratozoospermia does not have a significant impact on the CLBR of IVF treatment. What is known already There is ongoing debate regarding the clinical significance. Some studies indicate a pivotal role of sperm morphology in successful pregnancies, others cast doubt on its reliability as a predictive factor. Despite the application of stringent grading criteria, concerns remain about the subjective nature of assessing sperm morphology. Discrepancies in success rates between normozoospermic and isolated teratozoospermic groups shown by previous research findings underscore the uncertainty surrounding this matter. Inconsistent findings in previous research may be explained by limited sample sizes and a focus on immediate successes, neglecting subsequent frozen cycles. This study seeks to address the overlooked aspects of teratozoospermia. Study design, size, duration A retrospective study was conducted at Waterstone Clinic, involving 5254 IVF/ICSI autologous cycles completed between 2009-2022 cycles (completed meaning all embryos were either transferred or disposed of). Cycles were categorised according to sperm morphology and divided into two groups: the isolated teratospermic (ITS) group (1218 cycles) defined as having less than 4% normal morphologic forms with all other semen parameters being normal, and the normal sperm (NS) group (4036 cycles). Participants/materials, setting, methods The primary outcome measures, included fertilisation rate, number of cryopreserved embryos per egg collection, and CLBR. Employing multivariate statistics, the analysis explored the impact of interferons on CLBR in relation to variables such as sperm morphology, female age, egg number, and aetiology. Main results and the role of chance The female base characteristics were similar between the two groups in terms of age (37.1 vs 37.1 y/o), oocyte number (9.4 vs. 9.7) and FSH level (7.6 vs. 7.6 IU/L). Statistically significant differences were observed in fertilisation rates (61.7% vs. 65.8%, p < 0.001) and fertilisation failure (5.9% vs. 3.3%, p < 0.001) from IVF treatments when comparing the ITS and the NS groups. However, there was not any significant difference in the number of cryopreserved embryos between the two groups. Additionally, the CLBR did not show any significant difference between the two groups for IVF (35.8% vs. 37.8%, p > 0.05) or ICSI treatments (38.1% vs. 33.2%, p > 0.05). ICSI did not confer any benefit in improving the CLBR outcome when compared to conventional IVF, when adjusted for female age, number of oocytes collected, and aetiology. Limitations, reasons for caution While this study is retrospective it encompasses a substantial cohort. A potential for bias due to the selection criteria for IVF and ICSI exists. The intrinsic subjectivity in sperm morphology assessment introduces additional potential bias. Wider implications of the findings Despite the lack of support from CLBR to advocate for choosing ICSI in patients with ITS, this study suggests that couples with ITS experience lower fertilisation and higher fertilisation failure rates with IVF. This observation can serve as a valuable indicator for investigating the occurrence of fertilisation failure. Trial registration number not applicable

P-092 PGT-A Evaluates Advanced Sperm Selection in Embryos from High Teratozoospermia Males; a prospective randomized controlled trial

Abstract Study question Can sperm selection techniques improve the reproductive outcomes of ICSI patients, particularly in males with high teratozoospermia index (TZI) and sperm DNA fragmentation (SDF)? Summary answer Sperm selection techniques significantly increase fertilization and blastocyst development rates in males with high TZI and SDF, achieving a euploidy rate comparable to normal cases. What is known already Abnormal sperm morphology negative effects begin at the fertilization and reach the blastocyst quality and euploidy status. Men with abnormal teratozoospermic index (TZI) exhibit high levels of chromosomal abnormalities and sperm DNA fragmentation (SDF). SDF adversely affects pre and post-implantation embryo development events. Blastocyst aneuploidy isn’t solely due to the oocyte; but also, sperm contribution should be taken into consideration. Previous studies reported that advanced sperm selection techniques (PICSI and MACS) can efficiently enhance reproductive outcomes in abnormal SDF ICSI cases. The efficiency of these techniques in improving ICSI outcomes of patients with high TZI plus SDF remains unexplored. Study design, size, duration A prospective randomized controlled trial included a total of 264 couples, divided into three groups: (1) Control-1 group: males having normal TZI and no sperm selection techniques were used (n = 111), (2) Control-2 group (NO-SS): males having high TZI and no sperm selection techniques were used (n = 63), and (3) Sperm selection (SS) group: males having high TZI and SDF, and sperm selection techniques were used (n = 90). The study started from January till October 2023. Participants/materials, setting, methods Eligibility: females had ≥5MII oocytes, males had abnormal (>1.8) TZI, and all blastocysts (≥4BB) had trophoectoderm biopsy. Cases were randomized on the ICSI day to (control-2) and (SS) groups, along with (control-1) with normal TZI for comparison. Females were categorized into 2 subgroups (<35, and ≥35 years). SS techniques included physiological ICSI and magnetic-activated cell sorting. PGT-A was done using Illumina platform (Miseq). Statistical analysis (SPSS) of pre-implantation embryo development and PGT-A results was done. Main results and the role of chance We found no significant differences between the female, male ages and number of mature oocytes between the studied groups. Limitations, reasons for caution The results included all pre-implantation embryo parameters but, subsequent post-implantation events like implantation, pregnancy, and miscarriage rates will add more data about the competency of the resulting embryos. A Small sample size of females aged from 36 to 40 years old. Wider implications of the findings Sperm selection techniques appear to be efficient in improving fertilization, blastocyst development rates, and euploidy rates in males with high TZI and sperm DNA fragmentation, suggesting potential benefits for couples undergoing infertility treatment. Trial registration number NCT06231589

P-216 Whole genome amplification (WGA) of spent culture media may not be a reliable marker of blastocyst prioritization for transfer

Abstract Study question Does the success or failure of DNA amplification from spent culture media of good-quality blastocysts correlate with their ploidy status and clinical pregnancy outcomes? Summary answer Samples with successful DNA amplification exhibited similar rates of euploidy and clinical pregnancy rates compared to spent culture samples with failed DNA amplification. What is known already Studies have detected DNA in spent culture media/blastocoelic fluid (BF) from fully developed blastocysts. After amplification, this DNA is sequenced to provide information on their ploidy status. Some authors reported simply detection of amplified DNA as an indicator of blastocyst viability, so that spent culture media/BF with DNA amplification derive from abnormal embryos, and failed DNA amplification from euploid blastocysts. Along with this line of rationale, some authors associated DNA amplification failure with higher clinical pregnancy rates. Reproducibility of such findings is warranted which may stem from various factors, such as culture conditions, amplification methods and statuses of blastocysts analysed. Study design, size, duration This preliminary set of results is part of a cohort (nonselective) ongoing study, from which 62 patients participated so far from two tertiary IVF centers during the period of January 2022 until October 2023. Only ICSI patients aged <43-year-old with >6 mature oocytes were included. Single frozen blastocyst transfers were performed blinded to the results of the whole genome amplification (WGA) and/or DNA sequencing and pregnancy outcomes recorded. Participants/materials, setting, methods Embryos were singly cultured into 20µl GTL-media in Embryoscope+ incubator (Vitrolife). On day 3, embryos were washed repeated times and placed into new culture dish. Spent media was collected from ≥BL4BB blastocysts (Gardner) on days 5/6 after laser collapse prior vitrification. DNA from culture medium were amplified using MALBAC (Yikon-Genomics). After WGA, DNA was quantified using Qubit 3.0 fluorometer (cut-off of 0.6 ng/µl) and then subjected to next-generation sequencing (NGS) with Illumina MiSeq platform. Main results and the role of chance A total of 104 spent culture media samples were analysed from individual blastocysts. About 70% of samples of spent media had amplified DNA and were sequenced further. From those sequenced samples, 55% resulted in euploidy and 45% in abnormal ploidy status. Embryo transfers took place from blastocysts cultured in 82/104 samples. A total of 36 (44%) clinical pregnancies were obtained from 82 transfers/samples from which, 78% derived from samples with DNA amplification and 22% from samples with failed DNA amplification. Outcomes of clinical pregnancies were verified for both conditions, samples with DNA amplification or failed DNA amplification. A total of 28/55 (51%) samples with DNA amplification resulted in clinical pregnancies, while only 30% (8/27) of failed DNA amplification samples did so (P = 0.06). Positive predictive values (PPV) and negative predictive values (NPV) of ploidy status of sequenced samples in relation to ongoing pregnancy and live birth rates are ongoing. Limitations, reasons for caution The limited sample size. Wider implications of the findings The presence of DNA in the spent culture media may offer an attractive alternative as a non-invasive test for aneuploidy. Research is still ongoing and challenging. Nevertheless, truncating the test at DNA amplification phase seems not a reproducible methodology to serve as a criterion to prioritize embryos for transfer. Trial registration number This research project is financially supported by Theramex Birth Grant program, a competitive grant supporting IVF research in Europe.

Effect of Chronic Endometritis Treatment on The Outcome of ICSI in Patients with Repeated Implantation Failure

Effect of Chronic Endometritis Treatment on The Outcome of ICSI in Patients with Repeated Implantation Failure

Indications affect neonatal outcomes following early rescue ICSI: a retrospective study.

To investigate the impact of heterogeneity in patient indications or insemination protocols on neonatal outcomes of singletons following early rescue ICSI (rICSI) treatments. A retrospective study was conducted. Propensity score matching and multivariable logistic regression were used to adjust for confounders and biases. A total of 9095 IVF patients, 2063 ICSI patients, and 642 early rICSI patients were included in the study. No differences were detected in neonatal outcomes except small for gestational age (SGA) which increased in early rICSI patients compared with both unmatched and matched IVF groups with the risk ratio (RR) of 1.31 (95% CI: 1.05, 1.64) and 1.49 (95% CI: 1.05, 2.12). Further analysis showed that SGA increased significantly in partial fertilization failure (PFF) cycles with RRs of 1.56 (95% CI: 1.08, 2.27) and 1.78 (95% CI: 1.22, 2.59) compared with both unmatched and matched IVF patients but not in TFF patients. A positive association between fertilization rate via IVF and birth weight z-score was revealed in the PFF patients. Early rICSI in patients with total fertilization failure (TFF) appeared to be safe in terms of neonatal outcomes. However, when expanding the indications of rICSI to PFF patients, the SGA in the offspring increased, suggesting a potential effect on long-term health. Since other treatment options, such as using only the IVF-origin embryos still exist for these patients, further studies were needed to confirm the optimal decision for these patients.

Comparison of Hyaluronic Acid Binding and Conventional Sperm Selection For ICSI in Patients with Oligozoospermia Cohort

Comparison of Hyaluronic Acid Binding and Conventional Sperm Selection For ICSI in Patients with Oligozoospermia Cohort

O-178 Conventional IVF results in better embryo development when compared with ICSI in patients without severe male factor

Abstract Study question Which fertilisation technique offers better embryo development results in patients without severe male factor? Summary answer Conventional IVF offers better results regarding embryo development than ICSI in patients without male factor when applying both techniques in the same cohort of oocytes. What is known already ICSI was originally indicated for severe male factor. Nevertheless, it is currently used for many other indications so that it has become the most used technique. One of the reasons for the use of ICSI is the fear of a fertilisation failure when using conventional IVF (cIVF). We have to consider that ICSI is more expensive and invasive. For this reason, it is important to know whether there is a justification to use it in these cases. Study design, size, duration It is a randomized prospective study where 68 IVF/ICSI cycles were analysed. The study comprised 812 oocytes and 469 embryos and was conducted from January to December 2022 in the Assisted Reproduction Unit of a tertiary hospital. In each cycle, we performed a mixed technique using cIVF and ICSI in the same cohort of oocytes. MII rate, fertilisation rate and embryo development were analysed. Participants/materials, setting, methods All consecutive cycles with at least 6 fresh oocytes and semen parameters suitable for cIVF were offered to participate. In each cycle, oocytes were divided into two groups in order of collection and the fertilisation technique (cIVF or ICSI) was randomly assigned for each group. All the embryos were cultured to blastocyst stage. Data of maturity, fertilisation and embryo development were individually registered for every oocyte. Data were statistically analysed by applying multilevel regression models. Main results and the role of chance The patients average age was 36.3±3.0 (28 to 40) and the number of oocytes per cycle was 11.9±5.1 (6 to 26). 396 oocytes (48.8%) were allocated to cIVF group and 416 (51.2%) to ICSI group. The percentage of mature oocytes was significantly higher in cIVF group than in ICSI group (88.9% vs 81.5%, p = 0.017), which is consistent with the later assessment of maturity in cIVF. There were no statistically significant differences between cIVF and ICSI groups in terms of fertilisation rate (59.6% vs 56.0%) and high-quality blastocyst rate (A+B) (9.3% vs 8.7%) when analysed per oocyte. We observed clinical but not statistical differences in blastocyst rate per oocyte (30.6% cIVF vs 23.6% ICSI, p = 0.09). However, blastocyst rate per fertilised oocyte showed statistically significant differences (51.3% cIVF vs 41.6% ICSI, p = 0.047). Additionally, we found statistically significant differences between cIVF and ICSI in the rate of usable blastocysts (transferred or cryopreserved) per oocyte (26.3% vs 18.3%, p < 0.01) and per fertilised oocyte (44.1% vs 32.2%, p < 0.01). In 57.8% of the cycles, the best blastocyst was obtained by cIVF and in 42.2% by ICSI. Fertilisation failure was observed in 3 patients with cIVF and 1 patient with ICSI. All of them had ≤4 mature eggs. Limitations, reasons for caution This study includes the 68 patients who met the recruitment requirements until this moment. It is a part of a wider work, which will analyse a higher number of oocytes and pregnancy and perinatal outcomes. For this reason, the sample size is a limitation to the interpretation of the results. Wider implications of the findings Our findings encourage the use of cIVF when there is no severe male factor. Considering that cIVF is cheaper and less invasive than ICSI, the exclusive use of ICSI in these patients does not seem to be justified according to our results. Trial registration number not applicable

O-294 Dual trigger versus hCG-only trigger in ICSI patients: an analysis of 8500 cycles

Abstract Study question Among patients undergoing ovarian stimulation for ICSI, does dual trigger with hCG and GnRH agonist offer any benefit over mono trigger with hCG alone? Summary answer Dual trigger is associated with a higher number of mature oocytes without any impact on the pregnancy rate. What is known already In ART cycles, final oocyte maturation and resumption of meiosis are generally triggered by the administration of hCG as a surrogate for the natural LH surge. Several studies have investigated the role of a combination of a bolus of GnRH agonists and hCG to mimic the natural peak of endogenous LH and FSH, without clear answers regarding clinical benefit. A meta-analysis showed highernumbers of good-quality embryos and increased ongoing pregnancy rate after dual trigger and a recent systematic review demonstrated significantly higher live birth rate (LBR) per cycle after administration of dual trigger compared to hCG-only trigger. Study design, size, duration The study was a retrospective, single-centre cohort study. In this study, 8,525 ICSI cycles were included between January 2017 and April 2022 at a tertiary referral University Hospital. We included all patients undergoing ICSI in a GnRH antagonist ovarian stimulation cycle. The mono trigger consisted of recombinant or highly purified urinary hCG. In the dual trigger group, a bolus of GnRH agonist was combined with the hCG trigger. Participants/materials, setting, methods Our cohort of 8,525 cycles was divided into two groups: Group A, the mono trigger group, (7,022 cycles), and Group B, the dual trigger group (1503 cycles). Patients who underwent IVF, pre-implantation genetic testing, oocyte donation, and fertility preservation were excluded. Patients with uterine anomalies and endocrine disorders were also excluded, as well as patients who received a triptorelin-only trigger because of hyperresponse. Main results and the role of chance There was no difference in the mean age between the dual vs mono trigger cohorts (35.93 ± 4.90 vs 35.52±4.85). The most common indication for ART was male factor infertility in both groups (21.36% in group B vs 27.11% in group A). Stimulation was shorter in the dual trigger cohort (10.41 vs 12.08 p = 0.04). The total number of cumulus oocyte complexes (8.26±5.32 vs 7.45±4.68, p < 0.001), mature oocytes (6.39±4.15 vs 5.99±3.80, p = 0.006) and fertilized oocytes (4.79±3.59 vs 4.33±3.27, p = <0.001) was higher in the dual trigger group compared to those in the mono trigger group. Day 5 embryo transfer was more prevalent in the dual trigger group (46.07% vs 33,84%, p < 0.001). Embryo utilization rate was higher in group A (61.37% ±31.98) than in group B (53.47% ±30.91 p = <0.001). Ongoing regnancy rate was similar in both groups (27.29% in group A vs 27.08% in group B). Pearson chi2, Mann Whitney and logistic regression tests were used for the analysis. Limitations, reasons for caution In spite of the large sample, this study is retrospective and holds the possibility of unmeasured confounders. Wider implications of the findings Although a dual trigger may increase the number of mature oocytes, this does not translate into a higher pregnancy nor live birth rate. Reduced oocyte competence of the “surplus” oocytes may underlie this observation. Further research should identify specific subgroups that may benefit from dual trigger across meaningful outcome parameters. Trial registration number not applicable

Day after rescue ICSI: eliminating total fertilization failure after conventional IVF with high live birth rates following cryopreserved blastocyst transfer.

What is the impact of day after rescue ICSI (r-ICSI) on success of fresh and frozen embryo transfers? The use of r-ICSI can virtually allay fears of total fertilization failure (TFF) after conventional IVF (C-IVF) and achieve high live birth rates after frozen blastocyst transfer. More infertility clinics have resorted to the use of ICSI in place of C-IVF in IVF treatment owing to fear of TFF or a low fertilization rate. r-ICSI has been attempted either on the day of IVF or the day after. Day after r-ICSI has proved unsuccessful in the past. A retrospective data analysis was performed of 16 608 qualifying cases between April 2010 and July 2021 conducted at a single private academically affiliated fertility clinic. r-ICSI was performed principally on patients with >4 metaphase II oocytes, showing no signs of fertilization 18 h after C-IVF. C-IVF was performed on patients who had >4 million total motile sperm after preparation. r-ICSI was then performed 18-24 h after insemination, using the sperm sample from the previous day. r-ICSI fertilization rates, cryopreservation of cleavage and blastocysts embryos, and pregnancy rates after fresh or frozen transfer were then assessed. r-ICSI was performed on 377 patients (2.3% of eligible retrieval cycles) who had a mean (±SD) female and male age of 35.9 ± 4.5 and 38.1 ± 9.1 years, respectively. A total of 5459 oocytes were initially retrieved. Of the oocytes undergoing r-ICSI, 2389 (49.5%) fertilized normally, and 205 (54.4%) patients underwent a fresh embryo transfer. The live birth rates were 23/186 (12.3%) for fresh cleavage and 5/19 (26.3%) for fresh blastocyst stage transfers. In 145 cycles a blastocyst was frozen, and 137 transfers were performed with a 64/137 (46.7%) live birth rate. Of the 377 cycles receiving r-ICSI only, 25 of the qualifying cases failed to have any fertilization, reducing TFF to 25/16 608 (0.15%). This was a single-center retrospective study on a specific subset of patients, which may limit its generalizability to other clinics. r-ICSI allows a second opportunity to fertilize oocytes despite poor initial outcomes. Patients who had a frozen blastocyst transfer achieved high live birth rates, indicating that a resynchronization of the embryo with the endometrium can optimize r-ICSI cases. r-ICSI allays fears of TFF when using C-IVF, providing evidence that the overuse of ICSI in patients without male factor may not be warranted. The study was internally funded by Boston IVF. The authors declare that they have no conflict of interest in relation to the data published in the article. N/A.

PIEZO-ICSI increases fertilization rates compared with conventional ICSI in patients with poor prognosis.

Limited research has been published comparing PIEZO-ICSI with conventional ICSI. While positive effects have been documented in improving fertilization and degeneration, the outcomes in patients with previous poor results from conventional ICSI remain unclear. It is hypothesized that these patients may benefit the most from this form of insemination. This retrospective paired within-patient cohort study investigated patients (n=72) undertaking PIEZO-ICSI after a previous conventional ICSI cycle resulted in poor outcomes (including low fertilization (<50%), high degeneration (>15%), and/or poor embryo development and utilization). Patients required at least five oocytes collected in both cycles and a period of less than 2 years between the cycles. The outcomes of both cycles were compared in respect to fertilization, degeneration, embryo utilization, and pregnancy rates. Further analyses were applied to patients <38 and ≥38 years of age, with <50% or ≥50% fertilization with conventional ICSI and with <20% or ≥20% utilization with conventional ICSI. PIEZO-ICSI resulted in significantly higher fertilization (61.9% vs 45.3%, P<0.0001) and lower degeneration (7.7% vs 18.2%, P=0.0001) when compared to the conventional ICSI cycles. The greatest benefit was seen in patients who had less than 50% fertilization or <20% utilization in their conventional ICSI cycle, with improvements in fertilization and degeneration rates resulting in a significantly higher number of embryos utilized (frozen or transferred) per cycle. PIEZO-ICSI improved fertilization, degeneration, and utilization rates in patients with previous poor outcomes from conventional ICSI. The number of embryos available for use per cycle was also increased. Further significant improvements were achieved in patients who exhibited poor fertilization (<50%) or low utilization (<20%) from conventional ICSI.

Micro-TESE strategy in patients with NOA caused by AZFc deletion: synchronous or asynchronous?

In the treatment of infertile patients with non-obstructive azoospermia (NOA) caused by the deletion of the azoospermia factor c region (AZFc) on the Y chromosome, synchronous and asynchronous surgical strategies are discussed. Clinical data from NOA patients with the AZFc deletion who underwent micro-TESE were analyzed retrospectively. The sperm retrieval rate (SRR) and sperm utilization rate of synchronous and asynchronous operation groups were followed up and compared. The fertilization rate, high-quality embryo rate, clinical pregnancy rate, abortion rate, and cumulative live birth rate of ICSI in patients with successful sperm retrieval were compared between the two groups. The two groups had sperm utilization rates of 98.9% (93/94) and 50.0% (14/28), respectively. The asynchronous group's sperm consumption rates were much lower than those of the synchronous operation group. Fertilization rate, high-quality embryo rate, clinical pregnancy rate of fresh transfer cycle, abortion rate, and cumulative live birth rate of patients in the synchronous operation group with fresh sperm, and the asynchronous operation group with thawed sperm, respectively, were 30.6% vs 33.8%, 33.8% vs 40.7%, 40.0% vs 12.5%, 30.4% vs 7.1%. Between the two groups, there was no significant difference. This suggests that individuals with NOA caused by the AZFc deletion have a high possibility of successfully acquiring sperm using micro-TESE and ICSI to conceive their own offspring. Synchronous micro-TESE is recommended to improve sperm utilization rate and the cumulative live birth rate.

Evaluation of the role of three-dimensional ultrasound and power Doppler in prediction of endometrial receptivity in ICSI patients

Evaluation of the role of three-dimensional ultrasound and power Doppler in prediction of endometrial receptivity in ICSI patients

Evaluation of The Role of 3D Power Doppler in Predicting the Viability of The Endometrium to Receive Embryos for ICSI Patients

Evaluation of The Role of 3D Power Doppler in Predicting the Viability of The Endometrium to Receive Embryos for ICSI Patients

P-096 Clinical outcome of ICSI, PICSI and testicular spermatozoa-ICSI procedures in overcoming in patients with high level of sperm DNA fragmentation

Abstract Study question To compare the clinical effectiveness of ICSI, PICSI and testicular spermatozoa-ICSI (T-ICSI) in patients with abnormal level of sperm DNA fragmentation (SDF). Summary answer Clinical effect of T-ICSI procedure exceeds the effect of PICSI and ICSI in male infertility with high level of SDF. What is known already The effectiveness of ART in male infertility depends on sperm chromatin structure integrity. High level of SDF may be reason of IVF/ICSI procedure failures. Several studies have demonstrated impaired clinical outcomes of ICSI in patients with abnormal level of SDF. SDF has a positive correlation with pregnancy loss. It has been shown the benefit of using of testicular spermatozoa in ICSI in high level of SDF. PICSI is a method of the selection of sperm that may reduce the risk of reproductive losses. The data on the comparison of the effectiveness of these methods in high SDF are limited. Study design, size, duration Data was collected from 2019 to 2021 years. The couples with male factor of infertility were enrolled in the study. All the patients had abnormal level of SDF (30-45%). The age of women was less than 35 year. The couples were randomly selected and divided in 3 groups according the ART method: I (n = 25) ICSI; II (n = 22) PICSI; III (n = 20) T-ICSI procedure. Participants/materials, setting, methods The trial involved 67 couples. The mean patients age was 33,3±6,2 y (24-46), the women age - 29,1±2,3 (25-33). All of the men were non-smokers, had idiopathic pathospermia, high level of SDF and underwent unsuccessful courses of empirical therapy in anamnesis. SDF level was measured by TUNEL method. 52 (77,6%) couples had unsuccessful attempts of ICSI including pregnancy loss. The fertilization, blastocyst formation (Gr A/B), clinical pregnancy and live birth rates were measured. Main results and the role of chance It was not significant difference in SDF level in the groups: I group - 37,3±3,1%; II group - 37,2±3,5%; III group - 37,8±3,2% (III). The fertilization rate was comparable in the groups I, II and III - 65,4%; 66,6%; 63,3% respectively. Blastocyst formation (good quality embryos) rate was 41,2%; 46,1%; 56,1%(p &amp;lt; 0001) in I, II and III groups respectively. Clinical pregnancy and life birth rates were 36,0% [9/25]; 40,9%[9/22] (p &amp;lt; 0,05); 45,0%[9/20] (p &amp;lt; 0001) and 20,0% [5/25]; 27,2%[6/22]; 35%[7/20] in ICSI, PICSI, T-ICSI groups respectively. It was significant difference in miscarriage rate 16%, 13,7% (p &amp;lt; 005), 10% (p &amp;lt; 0001) in I, II и III groups respectively. Limitations, reasons for caution T-ICSI might compensate for ART failure in high level of SDF cases. Testicular sperm retrival is an invasive intervention, and its benefit must be carefully justified in each cases. Further studies are needed to determine whether infertility with sperm DNA damage can be overcomed by less invasive methods than T-ICSI. Wider implications of the findings There was no significant difference in fertilization rate in ICSI, PICSI and T-ICSI. T-ICSI demonstrated the best blastocyst formation rate and clinical outcome in trial. According results of our investigation PICSI might increase pregnancy rates and reduce pregnancy losses when compared with ICSI in infertility in high SDF. Trial registration number not applicable

O-043 ICSI: the gamechanger in ART

Abstract On behalf of Neelke De Munck, Herman Tournaye and all colleagues BrusselsIVF (UZBrussel) and Vrije Universiteit Brussel (1980-2022) In the early years of ART, IVF was successful to alleviate female-factor or idiopathic infertility, yet limited results were obtained in male-factor infertility. In the 80s, assisted fertilization procedures using micromanipulation were introduced, i.e. zona drilling and partial zona dissection. As these procedures did not perform well, at BrusselsIVF we investigated subzonal insemination (SUZI) in mice and eventually we introduced SUZI in our clinic for couples with impaired sperm parameters. IRB approval was obtained in 1991 but required a prospective follow-up protocol of all pregnancies and children born. During SUZI, it occurred that sperm was injected in the oolemma, after which normal fertilization and embryo development was observed. The first pregnancy leading to a live birth in January 1992 was established when 11 oocytes had no fertilization after SUZI but one oocyte was fertilized after “failed SUZI” and was eventually transferred. In the following period, we were able to demonstrate that ICSI was more efficient than SUZI and from July 1993 we only applied ICSI in patients in need of assisted fertilization. We deliberately choose to share our experience widely with our peers and thereafter ICSI was performed in many centres worldwide and ICSI has been mushrooming. The 2017 ICMART World Report mentions an annual increase of ART cycles with 2 million cycles in 2017 resulting in 330.000 deliveries. The percentage of cIVF versus ICSI is very different depending on the region but on average 70% of cycles are ICSI. This indicates that ICSI is also widely used for non-male infertility. The occurrence of about 15% failed fertilization cycles after cIVF is often the reason for using ICSI. This is for sure the case in countries where only a limited number of cycles is reimbursed by social security or where no reimbursement is in place. ICSI proved to relieve infertility in couples with severely impaired characteristics of ejaculated sperm including cryptozoospermia. For these patients ICSI results were similar as cIVF for other indications. ICSI with epididymal or testicular sperm could also solve the problem in patients with obstructive azoospermia. Patients with non-obstructive azoospermia can also be helped if sperm cells can be found by testicular sperm extraction using conventional TESE or microTESE. However, the chance to have a child in NOA couples embarking for both TESE and eventually ICSI is much lower if expressed in terms of Intention to Treat: 13,4% in NOA patients and 10,1% in Klinefelter patients. Most of these couples will embark on treatment even after full counselling before considering the use of donor sperm. At BrusselsIVF, ICSI is also used on in-vitro matured oocytes following culture of immature cumulus-oocyte complexes collected from antral follicles in PCOS patients. For couples undergoing Preimplantation Genetic Testing with biopsy of day-3 cleaving embryos or day-5 or day-6 blastocysts, ICSI is still the preferred procedure in order to avoid contamination at the time of biopsy by remnants of sperm or cumulus cells when cIVF is used. From the start in 1980 we have invested in clinical, translational and fundamental research. As of now this research is carried out by more than 250 individuals from BrusselsIVF, the centre for reproductive medicine of the UZBrussel and different research units at the VUB. Research focuses on different aspects of infertility, medical genetics, human reproduction, early human development and human pluripotent cells.

Marginal differences in preimplantation morphokinetics between conventional IVF and ICSI in patients with preimplantation genetic testing for aneuploidy (PGT-A): A sibling oocyte study.

ObjectiveThis study aimed to analyze the morphokinetic behaviour between conventional IVF and ICSI, in cycles with preimplantation genetic testing for aneuploidies (PGT-A).MaterialsA randomized controlled trial (NCT03708991) was conducted in a private fertility center. Thirty couples with non-male factor infertility were recruited between November 2018 and April 2019. A total of 568 sibling cumulus oocyte complexes were randomly inseminated with conventional IVF and ICSI and cultured in an Embryoscope time-lapse system. The morphokinetic behaviour of IVF/ICSI sibling oocytes was analysed as primary endpoint. As secondary endpoints, morphokinetic parameters that predict blastocysts that will be biopsied, the day of biopsy, gender and euploid outcome was assessed.ResultsWhen comparing IVF to ICSI, only the time to reach the 2-cell stage (t2) was significantly delayed for IVF embryos: OR: 1.282 [1.020–1.612], p = 0.033. After standardizing for tPNf (ct parameters), only Blast(tStartBlastulation-t2) remained significant: OR: 0.803 [0.648–0.994], p = 0.044. For the analysis of zygotes that will be biopsied on day 5/6 versus zygotes without biopsy, only early morphokinetic parameters were considered. All parameters were different in the multivariate model: ct2: OR: 0.840 [0.709–0.996], p = 0.045; ct6: OR: 0.943 [0.890–0.998], p = 0.043; cc2(t3-t2): OR: 1.148 [1.044–1.263], p = 0.004; cc3(t5-t3): OR: 1.177 [1.107–1.251], p<0.0001. When comparing the development between blastocysts biopsied on day 5 versus day 6, only three morphokinetic parameters were significant: cc2(t3-t2): OR: 1.394 [1.010–1.926], p = 0.044; ctBlastocyst: OR: 0.613 [0.489–0.768], p<0.0001 and ctExpandedBlastocyst: OR: 0.913 [0.868–0.960], p = 0.0004. Multivariate analysis of gender and ploidy did not reveal differences in morphokinetic behaviour.ConclusionMinor morphokinetic differences are observed between IVF and ICSI. Early in the development, distinct cleavage patterns are observed between embryos that will be biopsied or not.

Evaluation the effect of sildenafil on nitric oxide secretion and improvement of endometrial receptivity in fresh ICSI cycles

Introduction: Endometrial receptivity is a chain of events that promotes embryo implantation and is regulated by a variety of substances, such as cytokines, growth factors, and steroid hormones. The study&#39;s objective is to assess how vaginally delivered sildenafil in ICSI patients affects pregnancy outcome, endometrial thickness, subendometrial resistance index, and pulsatility index. Methods: The sixty women who participated in this controlled randomised trial (interventional experimental research) at the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies underwent IVF/ICSI cycles. Patients were split into two groups: Study group: From the day of the last menstrual period to the day of the hCG injection, thirty (30) infertile women got four (4) doses of vaginal sildenafil 25mg tablets daily. Thirty (30) infertile ladies who weren&#39;t given sildenafil therapy made up the control group. Results: Nitric oxide levels in the study group&#39;s serum were significantly significant. In the subendometrial Doppler research, the mean pulsatility index was considerably lower in the study group whereas the mean resistance index and mean systolic to diastolic ratio did not change statistically between the control and study groups. Between the control and study groups, there was no statistically significant difference in the mean endometrial thickness. The sildenafil group had a greater pregnancy rate, although the difference was not statistically significant. Conclusion: By enhancing subendometrial blood flow by lowering the pulsatility index and raising the nitric oxide (NO) factor, the use of vaginal sildenafil in a new ICSI cycle improves endometrial receptivity.

Ionophore application for artificial oocyte activation and its potential effect on morphokinetics: a sibling oocyte study

PurposeTo evaluate whether ionophore application at the oocyte stage changes the morphokinetics of the associated embryos in cases of artificial oocyte activation.MethodsIn a prospective sibling oocyte approach, 78 ICSI patients with suspected fertilization problems had half of their MII-oocytes treated with a ready-to-use ionophore (calcimycin) immediately following ICSI (study group). Untreated ICSI eggs served as the control group. Primary analyses focused on morphokinetic behavior and the presence of irregular cleavages. The rates of fertilization, utilization, pregnancy, and live birth rate were also evaluated.ResultsIonophore-treated oocytes showed a significantly earlier formation of pronuclei (t2PNa) and a better synchronized third cell cycle (s3) (P < .05). The rate of irregular cleavage was unaffected (P > .05). Ionophore treatment significantly improved the overall rates of fertilization (P < .01) and blastocyst utilization (P < .05).ConclusionIonophore application does not negatively affect cleavage timing nor is it associated with irregular cleavage.

P–324 Pregnancy outcome after IVF for endometriosis or male infertility: What differs?

Abstract Study question Are endometriosis women pregnant after IVF at increased risk of preeclampsia or placenta praevia than patients monitored for male infertility? Summary answer Patients with endometriosis are at greater risk than patients monitored for male infertility of developing preeclampsia and placenta previa. What is known already Endometriosis is a chronic estrogen-dependent disease that affects women of childbearing age which represents 10% of the general population[.The main symptoms found are chronic pelvic pain, infertility, dyspareunia and dysmenorrhea. Numerous publications have highlighted the deleterious effect of endometriosis on pregnancy i.e miscarriage, placental abnormalities, preeclampsia, preterm birth, low gestational weight. This complication may be related to the molecular and cellular abnormalities present in the endometrium of these patients and to the inflammatory state that may lead to abnormal contractility of the uterus at the time of the implantation window and trophoblastic invasion. Study design, size, duration This study is a retrospective, non-interventional monocentric cohort study conducted between January 2011 and December 2017 in Institut de Medecine de la Reproduction - Clinique Bouchard in Marseilles, France. Participants/materials, setting, methods The outcome of pregnancies obtained after IVF and/or ICSI in patients with endometriosis (n = 270) was compared with patients,free of endometriosis,monitored for male infertility (n = 366) The statistical study was carried out using GraphPad Version 8 The Student T-test was used to compare means across them. Results were considered significant for p &amp;lt; 0.05. Main results and the role of chance Patients with endometriosis and monitored during this period were older than those managed for male infertility. (33.59 vs 32.78) (p = 0.04). There was no difference between the two populations regarding BMI (p = 0.31) or smoking (p &amp;gt; 0.9). The rate of miscarriage observed in the two populations was comparable (25.37 vs. 25.78%) (p &amp;gt; 0.9), so was the rate of IUGR (5.81% vs. 2.29%) despite the observed percentages (p &amp;gt; 0.9). The rate of premature deliveries did not differ between the two populations (18.37% vs. 14.29%) (p = 0.55) neither did the number of children born with a weight &amp;lt;2500g at term (13.68% vs. 12.5%) (p = 0.83). Although the rate of gestational diabetes was comparable in both groups (4.11% vs 4.56%), the rate of preeclampsia was higher in the group of patients with endometriosis with a statistically significant difference (4.79% vs 0.79%) (p = 0.01). Similarly, the rate of placenta previa was higher in patients with endometriosis (4.11% vs 0.76%) (p = 0.02). All pregnancies complicated by placenta previa resulted from J2/J3 embryo transfer. Estradiol levels on the day of induction (2166 pg/ml vs 2452) (p = 0.67) and endometrial thickness was not different between patients with placenta praevia or no (10.45 vs 10.51) (p = 0.66). Limitations, reasons for caution Our study is retrospective which may introduce several biases despite the size of our sample i;e patients with endometriosis are older, adenomyosis was not included in the criteria. In our study we have not found any additional risk related to the type of embryo transferred. Wider implications of the findings: Patients with endometriosis are at greater risk than patients managed for male infertility of developing preeclampsia and placenta previa. It is advisable to warn patients of this possible complication, to promote e-SET and to set up early monitoring in order to place the appropriate management around these patients. Trial registration number Not applicable

Sperm Selection Procedures for Optimizing the Outcome of ICSI in Patients with NOA.

Retrieving spermatozoa from the testicles has been a great hope for patients with non-obstructive azoospermia (NOA), but relevant methods have not yet been developed to the level necessary to provide resolutions for all cases of NOA. Although performing testicular sperm extraction under microscopic magnification has increased sperm retrieval rates, in vitro selection and processing of quality sperm plays an essential role in the success of in vitro fertilization. Moreover, sperm cryopreservation is widely used in assisted reproductive technologies, whether for therapeutic purposes or for future fertility preservation. In recent years, there have been new developments using advanced technologies to freeze and preserve even very small numbers of sperm for which conventional techniques are inadequate. The present review provides an up-to-date summary of current strategies for maximizing sperm recovery from surgically obtained testicular samples and, as an extension, optimization of in vitro sperm processing techniques in the management of NOA.