Outcome Of Intracytoplasmic Sperm Injection Cycles Research Articles

P-510 The impact of the severe male factor in the egg donation cycles using preimplantation genetic test for aneuploidy

Abstract Study question Does severe male factor affect the euploidy rate and IVF outcomes in donor egg ICSI cycles using PGT-A? Summary answer Severe male factor affects the euploidy rate in egg donation cycles, however, no statistical differences were reported in live birth and cumulative live birth rate. What is known already Around 50% of couples seeking fertility treatment have male-factor infertility. Intracytoplasmic sperm injection (ICSI) relies on evaluating sperm morphology, but the direct relationship between semen quality and embryonic potential remains unclear. Evidence indicates that poor semen characteristics correlate with an increase in aneuploidy rate of spermatozoa. Therefore, pregenetic testing for aneuploidies (PGT-A) is recommended in cases of male factor infertility. The relationship between aneuploidy rates and abnormal sperm remains a debated topic. The use of the egg donation model offers a valuable chance to isolate the male-related factor and evaluate its influence on the occurrence of embryonic aneuploidies. Study design, size, duration This is a retrospective, observational, cohort study including all European IVI clinics. The study included egg donation cycles using pre-implantation genetic testing for aneuploidies (PGT-A) generated in IVI clinics from January 2017 until December 2023. 690 egg donation cycles were included in the study, N = 202 in the group with sperm < 5mil/ml, N = 102 in the sperm group with 5-15 mil/ml and N = 386 in the group with sperm >15 mil/ml. Participants/materials, setting, methods The study included women between 30-49 years undergoing egg-donor cycles, using fresh/frozen sperm Sperm samples were derived after 2-5 days of abstinence. The cycles of egg donation were divided into three groups according to sperm concentration, severe oligozoospermia (SO, < 5mil/ml), moderate oligozoospermia (MO, between 5-15 mil/ml), and control group(>15 mil/ml). Trophectoderm biopsy was performed on day 5 or 6 and subsequent analysis used the next sequencing generation (NGS). Single euploid blastocyst transfers were included. Main results and the role of chance The fertilization rate was decreased in SO (73.33 ± 16.19) than in the MO group (79.07 ±12.45) and control group (77.80 ±14.72) p = 0.0047. The number of blastocysts was higher in the control (5.80 ± 2.72) and MO (5.97± 2.78) than in the SO group (4.99 ± 2.40), p = <0.001. The number of euploid blastocysts was decreased in SO (3.21 ± 2.07) than in the control (4.09 ± 2.41) and MO group (4.05 ± 2.15), p = <0.001. The euploidy rate of SO decreased (63.32 ±26.39) compared to the control (69.67 ± 25.08) and MO group (68.55 ± 20.37), p = 0.012. The logistic regression model included possible confounders (male age, age donor, male BMI, sperm motility, sperm morphology and sperm concentration) that showed no effect on the euploidy rate. The pregnancy rate was similar across the groups 69.77% vs 66.05% vs 66.23%, p = 0.54. No statistical differences were reported for the clinical pregnancy rate 62.79% vs 61.73% vs 58.40%, p = 0.44, and the live birth rate 62.79% vs 61.73% vs 58.40%, p = 0.44. 48.05 vs 46.38% vs 42.64%, p = 0.37. No differences were reported in the miscarriage rate (p = 0.37) and CLBR (p = 0.26). Limitations, reasons for caution The retrospective nature of the study and the sample size represent the principal limitations of the study. The clinical reason for severe male factors was not considered in the study. Wider implications of the findings SO affects the quality affect the early embryonic potential through an increase in aneuploidy, despite no clinical effects.The use of PGT-A in case of severe male factor remains not justified in the egg donation.Future prospective multi-center studies are needed to highlight the effect of sperm quality on early embryonic potential. Trial registration number not applicable

Prediction of Reproductive Outcomes of Intracytoplasmic Sperm Injection Cycles Using a Multivariate Scoring System.

Prediction of in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI) success is crucial in counselling patients about their real chance of getting a live birth before commencing treatment. A multivariate scoring system proposed by Younis et al., 2010, was amongst the predictive models used to evaluate IVF/ICSI success. The score entitles basal endocrine, clinical and sonographic parameters. The objective of this study is to assess the predictability of the Younis multivariate score for pregnancy outcomes in ICSI cycles. This prospective observational cohort study (NCT03846388) included patients who pursued IVF or ICSI in a tertiary infertility unit between February 2019 and December 2021. The score variables were age, body mass index, antral follicle count, basal follicle-stimulating hormone (FSH), basal FSH/luteinising hormone ratio, infertility duration, number of previous cancellations and mean ovarian volume. For each woman included in the study, Younis multivariate score was calculated. Then, we correlate the different reproductive outcomes with score levels to validate the score predictability. A score of ≤14 was defined as a low score based on the previous study's results. The student's t-test and Mann-Whitney test were used to compare numerical variables, whereas categorical variables were analysed using the Chi-square test. A receiver operating curve (ROC) and a multivariate logistic regression model were used to investigate the predictability of the Younis scoring model for cycle outcomes. Two hundred ninety-two ICSI-ET cycles were analysed. Of the total cohort, 143 (48.97%) women included showed a low score (≤14), whereas 149 (51.03%) women showed a high score (>14). Women with low scores had significantly higher pregnancy and live birth rates compared to women with high scores (60.1% vs. 7.4%, respectively, P < 0.001; 44.7% vs. 6.7%, respectively, P < 0.001). The area under the curve (AUC) in the ROC curve analysis showed a higher predictability for the scoring system for live birth rate with an AUC of 0.796, with a sensitivity of 86.5% and specificity of 63.8% when using a cut-off level of ≤14. For pregnancy prediction, the AUC was 0.829, with a sensitivity of 88.66% and a specificity of 70.77% when using the same cut-off. Women who have a low score have a high chance of having frozen embryos. Likewise, women who have a high score have a very high chance of cycle cancellation. The Younis multivariate score can be used for the prediction of ICSI cycle outcomes and to calculate the chance of cycle cancellation, pregnancy and take-home baby before ICSI.

Y chromosome microdeletions in Chinese men with infertility: prevalence, phenotypes, and intracytoplasmic sperm injection outcomes

BackgroundThe incidence of Y chromosome microdeletions varies among men with infertility across regions and ethnicities worldwide. However, comprehensive epidemiological studies on Y chromosome microdeletions in Chinese men with infertility are lacking. We aimed to investigate Y chromosome microdeletions prevalence among Chinese men with infertility and its correlation with intracytoplasmic sperm injection (ICSI) outcomes.MethodsThis single-center retrospective study included 4,714 men with infertility who were evaluated at the Reproductive Center of the First Affiliated Hospital of Sun Yat-sen University between May 2017 and January 2021. Semen analysis and Y-chromosome microdeletion via multiplex polymerase chain reaction were conducted on the men. The study compared outcomes of 36 ICSI cycles from couples with male azoospermia factor (AZF)cd deletions with those of a control group, which included 72 ICSI cycles from couples without male Y chromosome microdeletions, during the same period. Both groups underwent ICSI treatment using ejaculated sperm.ResultsAmong 4,714 Chinese men with infertility, 3.31% had Y chromosome microdeletions. The combined deletion of sY254 and sY255 in the AZFc region and sY152 in the AZFd region was the prevalent pattern of Y chromosome microdeletion, with 3.05% detection rate. The detection rates of AZF deletions in patients with normal total sperm count, mild oligozoospermia, severe oligozoospermia, cryptozoospermia, and azoospermia were 0.17%, 1.13%, 5.53%, 71.43%, and 7.54%, respectively. Compared with the control group, the AZFcd deletion group exhibited no significant difference in the laboratory results or pregnancy outcomes of ICSI cycles using ejaculated sperm.ConclusionsThis is the largest epidemiological study on Y chromosome microdeletions in Chinese men with infertility. The study results underline the necessity for detecting Y chromosome microdeletion in men with infertility and severe sperm count abnormalities, especially those with cryptozoospermia. The combined deletion of sY254 and sY255 in the AZFc region and sY152 in the AZFd region was the most prevalent Y chromosome microdeletion pattern. Among patients with AZFcd deletion and ejaculated sperm, ICSI treatment can result in pregnancy outcomes, similar to those without AZFcd deletion.

Effect of ejaculatory abstinence period on fertilization and clinical outcomes in ICSI cycles: a retrospective analysis

Does ejaculatory abstinence impact fertilization outcomes in intracytoplasmic sperm injection (ICSI) cycles in infertile couples? This single-centre retrospective observational study included 6919 ICSI cycles from 2013 to 2022. The primary outcome was the assessment of oocyte fertilization, measured in terms of the rate of formation of two-pronuclear (2PN), 3PN and 1PN zygotes. Secondary outcomes were blastulation, cumulative positive β-human chorionic gonadotrophin test and clinical pregnancy rates. Relationships between ejaculatory abstinence and fertilization outcomes, and ejaculatory abstinence and clinical outcomes were evaluated with multivariable analysis, including possible confounders. A positive association was observed between ejaculatory abstinence and semen sample volume (P < 0.001), sperm concentration (P < 0.001) and total motile sperm count (P < 0.001). No association was found between the 1PN zygote rate and ejaculatory abstinence (P = 0.97). Conversely, for each additional day of ejaculatory abstinence, the likelihood of obtaining 2PN zygotes from all inseminated oocytes decreased by 3% [adjusted odds ratio (aOR) 0.97, 95% CI 0.94-0.99], whilst the likelihood of obtaining 3PN zygotes from all inseminated oocytes increased significantly by 14% (aOR 1.14, 95% CI 1.07-1.22). No significant associations were found between ejaculatory abstinence and blastulation, cumulative pregnancy or miscarriage rates. A longer ejaculatory abstinence period significantly decreases the rate of 2PN zygotes, and increases the rate of 3PN zygotes without directly affect blastulation and pregnancy rates.

A 3–4 h oocyte retrieval-ICSI interval optimizes clinical outcomes for women over 40 years

To explore an appropriate time interval between oocyte retrieval and intracytoplasmic sperm injection (ICSI) for optimal embryological and clinical outcomes in ICSI cycles over 40 years of maternal age. A retrospective analysis of 1476 ICSI fresh cycles from women aged over 40 years, was performed at the Reproduction Medicine Research Center of the Sixth Affiliated Hospital of Sun Yat-sen University, between December 2013 and August 2020. The fertilization rate and clinical pregnancy rate were the main outcomes. Multivariate linear regression and logistic regression analysis of factors showed that fertilization rate (P = 0.024) and clinical pregnancy rate (P = 0.011) were significantly associated with oocyte pick up (OPU)-ICSI interval. A longer OPU-ICSI interval (no more than 6 h) was associated with a higher fertilization rate but significantly decreased the clinical pregnancy rate when the OPU-ICSI interval was over 4 h (P < 0.05). The optimal OPU-ICSI interval is between 3 and 4 h for excellent embryological and clinical outcomes in ICSI cycles over 40 years of maternal age.

The impact of female BMI on sperm DNA damage repair ability of oocytes and early embryonic development potential in intracytoplasmic sperm injection cycles.

Obesity adversely influences the quality of oocytes and embryos and can affect DNA repair in embryos, leading to reproductive issues. However, the effects of body mass index (BMI) on DNA repair ability in oocytes during intracytoplasmic sperm injection (ICSI) cycles have not yet been investigated. Therefore, this retrospective study aimed to analyze the influence of sperm DNA damage on embryo development and reproductive outcomes in overweight/obese and normal-weight women in ICSI cycles. A total of 1,141 patients who received the first fresh ICSI cycle treatments were recruited from July 2017 to July 2021. Based on the BMI of the women, all patients were divided into normal weight (18.5≤BMI<25 kg/m2; n=824; 72.22%) and overweight/obese (BMI≥25 kg/m2; n=317; 27.78%) groups. Furthermore, according to the sperm DNA fragmentation index (DFI), these two groups were subdivided into two subgroups: DFI<30% and DFI≥30%. In the normal-weight women group, the embryonic development and reproductive outcomes of ICSI cycles were not statistically different between the two subgroups (DFI<30% and DFI≥30%). However, in the overweight/obese women group, couples with a sperm DFI≥30% had a significantly lower fertilization rate (76% vs. 72.7%; p=0.027), cleavage rate (98.7% vs. 97.2%; p=0.006), and high-quality embryo rate (67.8% vs. 62.6%; p=0.006) than couples with a sperm DFI<30%. When injected sperm with high DFI into the oocytes of overweight/obese women, resulting in lower fertilization, cleavage, and high-quality embryo rates in ICSI cycles, and the decreased early developmental potential of embryos from overweight/obese patients may be caused by the diminished capacity of oocytes to repair sperm DNA damage.

Telomere Length in Human Spermatogenic Cells as a New Potential Predictor of Clinical Outcomes in ART Treatment with Intracytoplasmic Injection of Testicular Spermatozoa.

Predicting the clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles that use the testicular spermatozoa of azoospermic patients presents a challenge. Thus, the development of additional approaches to assessing the competence of a testicular-sperm-derived embryo without causing damage to gametes or the embryo is necessary. One of the key parameters in determining such developmental competence is telomere length (TL). We aimed to analyze TLs in spermatogenic cells from the testicular biopsy samples of azoospermic patients and determine how this parameter influences embryo competence for pre- and post-implantation development. Using Q-FISH, we studied the TL of the chromosomes in spermatogonia and spermatocytes I from the TESE biopsy samples of 30 azoospermic patients. An increase in TL was detected during the differentiation from spermatogonia to spermatocytes I. The patients' testicular spermatozoa were used in 37 ICSI cycles that resulted in 22 embryo transfers. Nine pregnancies resulted, of which, one was ectopic and eight ended in birth. The analysis of embryological outcomes revealed a dependence between embryo competence for development to the blastocyst stage and the TL in spermatogenic cells. The TLs in spermatogonia and spermatocytes I in the testicular biopsy samples were found to be higher in patients whose testicular sperm ICSI cycles resulted in a birth. Therefore, the length of telomeres in spermatogenic cells can be considered as a potential prognostic criterion in assessing the competence of testicular-sperm-derived embryos for pre- and post-implantation development. The results of this study provide the basis for the development of a laboratory test for the prediction of testicular sperm ICSI cycle outcomes.

Comparison of Reproductive Outcomes in ICSI Cycles Using Sperm Chip Technique and Density Gradient Technique in Men with Normal Semen Analysis

Background: Microfluidic sperm sorting procedure or sperm chip technique is a chemical-free method of selecting sperm using a disposable chip. It is a new gentle alternative for sperm processing which has been produced to obtain sperm with higher rates of motility and better morphology, as well as to reduce DNA fragmentation in sperm with high DNA fragmentation to nearly undetectable levels. We aimed to evaluate sperm chip techniques on clinical pregnancy rates in patients who underwent intracytoplasmic sperm injection (ICSI). Methods: The patients in whom fresh embryo transfer (ET) on Day-3 or 5 after ICSI had been performed were analyzed in this prospective randomized cohort study. Results: Of those, 102 patients underwent ICSI with sperm isolated using sperm chip technique (study group) while 111 patients underwent ICSI with sperm isolated using swim-up technique (control group). No significant difference was observed between the groups in terms of fertilization rate in patients who underwent ET on the 3rd or 5th day. In the patients having ET on Day-3, Grade 1 embryos were obtained similarly between the sperm chip group and the control group. Grade 1 embryos transferred on Day-5 were observed significantly more frequently in the study group (p = 0.050). However, clinical pregnancy rates did not show significant differences between the groups in patients who were transferred on both the 3rd and 5th days. Conclusions: Although sperm selection using by sperm chip technique provides advantage in terms of blastocyst quality, use of this technique does not enhance success in terms of clinical pregnancy. Clinical Trial Registration: Approval was obtained from ClinicalTrials.gov with NCT03355937 approval number.

Profiling the male germline genome to unravel its reproductive potential

To identify specific germline mutations related to sperm reproductive competence, in couples with unexplained infertility. In this retrospective study, couples were divided according to whether they had successful intracytoplasmic sperm injection outcomes (fertile) or not (infertile). Ancillary sperm function tests were performed on ejaculates, and whole exome sequencing was performed on spermatozoal DNA. Sperm aneuploidy and gene mutation profiles were compared between the 2 cohorts as well as according to the specific reasons for reproductive failure. Center for reproductive medicine at a major academic medical center. Thirty-one couples with negative infertility workups and normal semen parameters. Couples with mutations on fertilization- or embryo development-related genes were subsequently treated by assisted gamete treatment or microfluidics, respectively. Intracytoplasmic sperm injection cycle outcomes including fertilization, clinical pregnancy, and delivery rates. Sperm aneuploidy was lower in the fertile group (4.0% vs. 8.4%). Spermatozoa from both cohorts displayed mutations associated with sperm-egg fusion (ADAM3A) and acrosomal development (SPACA1), regardless of reproductive outcome. The infertile cohort was then categorized according to the reasons for reproductive failure: absent fertilization, poor early embryo development, implantation failure, or pregnancy loss. Spermatozoa from the fertilization failure subgroup (n = 4) had negligible PLCζ presence (10% ± 9%) and gene mutations (PLCZ1, PIWIL1, ADAM15) indicating a sperm-related oocyte-activating deficiency. These couples were successfully treated by assisted gamete treatment in their subsequent cycles. Spermatozoa from the poor early embryo development subgroup (n = 5) had abnormal centrosomes (45.9% ± 5%), and displayed mutations impacting centrosome integrity (HAUS1) and spindle/microtubular stabilization (KIF4A, XRN1). Microfluidic sperm processing subsequently yielded a term pregnancy. Spermatozoa from the implantation failure subgroup (n = 7) also had abnormal centrosomes (53.1% ± 13%) and carried mutations affecting embryonic implantation (IL9R) and microtubule and centrosomal integrity (MAP1S, SUPT5H, PLK4), whereas those from the pregnancy loss subgroup (n = 5) displayed mutations on genes involved in trophoblast development (NLRP7), cell cycle regulation (MARK4, TRIP13, DAB2IP, KIF1C), and recurrent miscarriage (TP53). By assessing the sperm genome, we identified specific germline mutations related to various reproductive processes. This information may clarify elusive factors underlying reproductive competence and enhance treatment for couples with unexplained infertility.

Assessing male gamete genome integrity to ameliorate poor assisted reproductive technology clinical outcome

To assess the role of evaluating sperm chromatin fragmentation (SCF) as a tool to guide treatment in couples who achieved unexpectedly poor clinical outcomes after intracytoplasmic sperm injection (ICSI). We identified couples with an unexpectedly suboptimal clinical outcome after ICSI who were then screened for SCF. Consequently, the same couples were counseled to undergo a subsequent ICSI cycle using either ejaculates processed by microfluidic sperm selection (MFSS) or spermatozoa retrieved from the testis, and clinical outcomes were compared between history and treatment cycles. To confirm the sole effect of a compromised male gamete, we compared the ICSI outcome in cycles where male gametes with abnormal SCF were used to inseminate autologous and donor oocytes. Finally, to eliminate an eventual confounding female factor component, we compared the clinical outcome of ICSI cycles using sibling donor oocytes injected with spermatozoa with normal or abnormal SCF. Academic reproductive medicine center point of care. The patient population consisted of 76 couples with reproductively healthy and relatively young female partners and male partners with compromised semen parameters, but suitable for ICSI. In a subanalysis, we identified 67 couples with abnormal SCF who underwent ICSI cycle(s) with donor oocytes. Furthermore, we identified 29 couples, 12 with normal SCF and 17 with abnormal, uncorrected SCF, and 7 couples with abnormal, corrected SCF vs. a control, who used sibling donor oocytes for their ICSI cycle(s). For couples who resulted in surprisingly low clinical outcomes after ICSI, despite semen parameters adequate for ICSI and a normal female infertility evaluation, a SCF assessment was performed on the semen specimen using the terminal deoxynucleotidyl transferase-mediated fluorescein-deoxyuridine triphosphate nick-end labeling (TUNEL) assay. The couples then underwent a subsequent ICSI cycle with spermatozoa processed by MFSS or surgically retrieved. Moreover, cycles with donor oocytes were used to confirm the sole contribution of the male gamete. Clinical outcomes, such as fertilization, embryo implantation, clinical pregnancy, delivery, and pregnancy loss rates were compared between history and treatment cycle(s) using ejaculated spermatozoa selected by MFSS or from a testicular biopsy, taking into consideration the level of SCF. In a subanalysis, we reported the clinical outcomes of 67 patients who used donor oocytes and compared them with cycles where their own oocytes were used. Furthermore, we compared the ICSI clinical outcomes between cycles using sibling donor oocytes injected with low or high SCF with or without sperm intervention aimed at correcting, or alleviating the degree of SCF. In a total of 168 cycles, 76 couples had in a prior cycle a 67.1% fertilization rate, and clinical pregnancy and pregnancy loss rates of 16.6% and 52.3%, respectively. After testing for SCF, the DNA fragmentation rate was 21.6%. This led to a subsequent ICSI cycle with MFSS or testicular sperm extraction, resulting in clinical pregnancy and delivery rates of 39.2%, and 37.3%, respectively. The embryo implantation rate increased to 23.5%, whereas the pregnancy loss rate decreased to 5% in the treatment cycle. This was particularly significant in the moderate SCF group, reaching embryo implantation, clinical pregnancy, and delivery rates of 24.3%, 40.4%, and 36.2%, respectively, and reducing the pregnancy loss rate to 10.5% in post-sperm treatment cycles. In 67 patients with high SCF who used donor oocytes, a significantly higher fertilization rate of 78.1% and embryo implantation rate of 29.1% were reported, compared with those in couples also with an elevated SCF who used their own. Interestingly, the clinical pregnancy and delivery rates only increased slightly from 28.0%-36.1% and from 23.7%-29.2%, respectively. To further control for a female factor, we observed couples who shared sibling donor oocytes, 17 with normal SCF and 12 with abnormal (uncorrected) SCF. Interestingly, the abnormal SCF group had impaired fertilization (69.3%), embryo implantation (15.0%), and delivery (15.4%) rates. For an additional 15 couples who split their donor oocytes, 8 had normal SCF, and although 7 couples originally had abnormal SCF, 4 used microfluidic processing, 2 used testicular spermatozoa, and 1 used donor spermatozoa to alleviate the degree of SCF, resulting in comparable clinical outcomes with the normal SCF group. A superimposed male factor component may explain the disappointing ICSI outcome in some couples despite reproductively healthy female partners. Therefore, it may be useful to screen couples for SCF to guide treatment options and maximize chances of a successful pregnancy. The improved, but suboptimal pregnancy and delivery outcomes observed in couples using donor oocytes confirmed the exclusive detrimental role that the male gamete exerted on embryo development despite the presence of putative oocyte repair mechanisms.

O-296 Paternal age does not affect obstetric and perinatal outcomes in IVF or ICSI cycles with autologous oocytes

Abstract Study question Does paternal age have any effect on obstetric and perinatal outcomes in in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) cycles with autologous oocytes? Summary answer Gestational diabetes, delivery type, cranial circumference and preterm birth were significantly different between our age groups, although after adjusted analysis no significant differences were seen. What is known already In recent years, there has been an increase of the average male age at which the first child is conceived. Therefore, studies addressing paternal age and reproductive risks are increasing and results are controversial, so there is a need of more research about it, which this study tries to address. Although several studies suggest the threshold to consider paternal age ‘advanced’ at 40 years, this limit is not well established. The assessment of risks concerning the mother and offspring is important and, thus, so is the study of its association with paternal age. Study design, size, duration This retrospective observational multicentric cohort study has considered pregnancies and children conceived from IVF-ICSI performed to couples in Spain IVIRMA clinics between January 2008 and March 2020 using patients’ own sperm and autologous oocytes. Paternal age ranged from 18 to 51 years. The study population was categorized in three groups: &amp;lt;30(A, reference), 30-40(B) and &amp;gt;40(C) years. The data available included 14402 patients who had a delivery with a live birth and 16118 singleton newborns. Participants/materials, setting, methods We evaluated pregnancies and children from autologous IVF-ICSI with own semen, known age and resulting in delivery. Data was exported to obtain the clinical database followed by the statistical analysis of the study main outcomes. P &amp;lt; 0.05 was considered statistically significant. Obstetric and perinatal outcomes were measured including delivery type, incidence of preterm birth, gestational diabetes, anemia, hypertension and pre-eclampsia; and newborn sex, weight, length, cranial circumference, Apgar score and neonatal intensive care unit (NICU) admission. Main results and the role of chance Results are expressed as rate or mean with 95%CI. There were no statistically significant differences for the incidence of anemia, hypertension or pre-eclampsia; newborn sex, weight, length, Apgar score (1, 5 and 10 min) and NICU admission between the reference younger group (A) vs. the older paternal age groups (B and C). However, there were statistically significant differences for the incidence of gestational diabetes when comparing A 5.1%(3.5-7.2) with B 7.7%(7.1-8.3) with OR = 1.5(1.1-2.2)(p = 0.021) and with C 9.1%(7.1-11.4) with OR = 1.9(1.2-2.9)(p = 0.005); for delivery type when comparing A[cesarean 33.8%(31.3-36.4)] with B[cesarean 37.8%(37.0-38.7)] with OR = 1.2(1.1-1.3) (p = 0.004) and with C[cesarean 49.3%(46.7-52.0)] with OR = 1.9(1.6-2.2) (p &amp;lt; 0.001); for cranial circumference when comparing A 34.5(34.3-34.7) with C 34.8(34.6-35.0) with p = 0.03; and for the incidence of preterm birth when comparing A 7.9%(6.6-9.5) with C 5.9%(4.7-7.2) with OR = 0.7(0.5-1.0)(p = 0.034). These results suggest an increased risk of gestational diabetes and cesarean delivery as paternal age increases. Moreover, group C showed a larger cranial perimeter and a lower risk of preterm birth when compared to group A. However, after adjusted analysis (according to selected patients’, cycles’ and semen samples’ characteristics, gestational age, newborn sex and type of delivery) these differences were no longer statistically significant, so probably other clinical variables are involved in the differences seen. Limitations, reasons for caution Despite the retrospective nature of this study that leads to biases derived from the clinical practice and to the presence of missing data limiting sample size, the study includes a huge sample size. Wider implications of the findings Currently, fatherhood delay has led to a growing interest in studies addressing paternal age and reproductive risks. Our paternal age comparison showed significant differences for gestational diabetes, delivery type, cranial circumference and preterm, no longer significant after adjusted analysis, so probably other clinical variables were involved. Further studies are needed. Trial registration number Not applicable

O-134 Predictive value of seminal oxidation-reduction potential (ORP) and sperm DNA fragmentation (SDF) analysis for reproductive outcomes of intracytoplasmic sperm injection (ICSI) cycles

Abstract Study question Do seminal oxidation-reduction potential (ORP) and sperm DNA fragmentation (SDF) predict ICSI cycle outcomes? Summary answer Both SDF and seminal ORP have strong prognostic value in predicting good fertilization (≥80%), blastocyst development (≥60%), clinical pregnancy and live birth. What is known already Seminal oxidative stress (OS) has been found to play an important role in various aetiologies of male infertility and is induced by the imbalance between reactive oxygen species (ROS) and antioxidants. OS is a major cause of sperm DNA fragmentation (SDF). It is well established that high levels of seminal OS and SDF are negatively associated with ART outcomes. Previous studies have associated high levels of SDF with an increase risk of miscarriage and a decrease in live birth rate. Study design, size, duration This prospective study included 144 couples who had a fresh autologous ICSI cycle between June 2018 and December 2020. There was no restriction to patients with severe male factor infertility. All cryopreservation and third party assisted reproductive techniques were excluded. Participants/materials, setting, methods This multi-centre study was performed at three ART Clinics in Cape Town, South Africa. All couples included in the study had fresh autologous ICSI cycles with freshly ejaculated semen. Seminal ORP testing was performed using the MiOXSYS system and SDF by means of a fluorescence microscopic TUNEL assay on freshly ejaculated semen samples that were used for ICSI. The ART outcomes evaluated were fertilization rate, blastocyst development rate, clinical pregnancy rate and live birth rate. Main results and the role of chance The study shows that seminal ORP significantly negatively correlates with fertilization rate (r=-0.267; P = 0.0012), blastocyst development rate (r=-0.389; P &amp;lt; 0.0001), pregnancy (r=-0.296; P = 0.0003) and live birth (r=-0.353; P &amp;lt; 0.0001). ROC curve analysis shows significant predictive power for ORP for fertilization (≥80%; AUC=0.652; P = 0.0012), blastocyst development rate (≥60%; AUC=0.762; P &amp;lt; 0.0001), pregnancy (AUC=0.677; P = 0.0002) and live birth (AUC=0.729; P &amp;lt; 0.0001). Comparable results were obtained for SDF. An average cut-off value for ORP of 0.55 mV/106 sperm/mL was calculated. The AUCs between the SDF and ORP results for all reproductive outcome parameters did not differ. For all reproductive outcome parameters, normal sperm morphology shows the lowest predictive power. ORP with male age as confounding factor had significant effects on odds ratios (OR) of fertilization (OR = 0.1446; CI: 0.048-0.430; P = 0.0001) and blastocyst development (OR = 0.2468; CI: 0.095-0.635; P = 0.0029), while it was not significant for pregnancy and life birth. Multivariate logistic regression with fertilization, blastocyst development, pregnancy and live birth as dependent parameters and high/low seminal ORP, primary and secondary infertility, unexplained infertility, polycystic ovaries, endometriosis, tubal factor myomectomy, ovarian insufficiency and advanced maternal age as independent variables showed overall model fits (P &amp;lt; 0.0002) with a significant (P &amp;lt; 0.0084) influence of ORP on all reproductive outcome parameters. Limitations, reasons for caution Since only ICSI patients were included in this study, this might have contributed to the fact the cut-off value of ORP is lower than the published cut-off. A further limitation is the use of a fluorescence microscopic TUNEL assay which drastically limited the number of sperm evaluated. Wider implications of the findings This is the first study showing predictive value of seminal ORP for reproductive outcome including pregnancy and live birth. The method is easy and much quicker than current methods for the determination of SDF. Hence, it can be used a point of care method. Trial registration number Not applicable

Are Clinical Outcomes of Micro-TESE in Non-obstructive Azoospermic Men Affected by the Use of Fresh or Frozen Gametes?

Background: The combination of microsurgical testicular sperm extraction (mTESE) and intracytoplasmic sperm injection (ICSI) has become a common management option, with sperm motility being positively associated with successful outcomes. However, few studies have investigated whether the use of fresh or thawed gamete combinations affect clinical outcomes. Objectives: To determine whether the clinical outcomes of ICSI cycles using mTESE recovered testicular sperm of non-obstructive azoospermia (NOA) patients are affected by using fresh or thawed gametes. Material and Methods: A retrospective study was conducted of NOA patients who underwent mTESE between 2017 and 2020 at Monash IVF assisted reproductive clinics in Melbourne, Australia. The impact of gamete fresh/frozen status and sperm motility was investigated on clinical outcomes such as fertilization, blastocyst formation, clinical pregnancy, and live birth rates (LBRs). Results:A total of 103 NOA patients underwent mTESE, with a 65.1% successful surgical-sperm-retrieval. In total 56 patients contributed to 68 ICSI cycles, with a 35.1% fertilization rate and 25% LBR per embryo transfer. Compared with fresh testicular sperm, thawed testicular sperm did not affect clinical outcomes, including LBRs [16.7% vs 12.0%; odds ratio (OR) 0.68 (0.18–2.70)]. However, the use of thawed oocytes had a negative effect on fertilization rates [fresh-oocytes, 37.8%; vitrified-oocytes, 34.5%; OR 0.86 (0.02–0.48)]. Cycles using only motile sperm had a greater fertilization rate than those using a combination of motile and non-motile sperm (49.6% vs 37.2%, p ¡ 0.05). Importantly, when exclusively non-motile sperm were available (n = 26 cycles) their injection resulted in a very low fertilization rate (2.7%) and no live births were recorded. Conclusion: Micro-TESE is an effective treatment for NOA patients, with no clear advantage of using fresh over thawed sperm, however, the use of vitrified compared with fresh oocytes requires further investigation. Importantly, patients should be informed of the poor outcomes with the use of non-motile sperm in mTESE ICSI cycles.

Does sperm DNA fragmentation affect clinical outcomes during vitrified-warmed single-blastocyst transfer cycles? A retrospective analysis of 2034 vitrified-warmed single-blastocyst transfer cycles.

Sperm DNA is essential in embryo development. The sperm DNA fragmentation index (DFI), which reflects the degree of sperm DNA fragmentation (SDF), is a crucial biomarker in evaluating the sperm quality. However, whether SDF influences the clinical outcomes after in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) remains controversial. This study aimed to investigate the relationship between sperm DNA SDF and clinical outcomes of vitrified-warmed single-blastocyst transfer cycles. A total of 2034 vitrified-warmed single-blastocyst transfer cycles (536 from ICSI and 1498 from IVF) were included in this analysis. According to the sperm DFI, all cycles were divided into two groups (DFI < 27.3% group and DFI ≥ 27.3% group). The Mann-Whitney and chi-squared tests were used to compare patient characteristics and clinical outcomes between the two groups. Furthermore, logistic regression analysis was performed to analyze the association between SDF and clinical outcomes. The chi-squared test showed no differences in positive human chorionic gonadotropin (HCG) rate, clinical pregnancy rate, miscarriage rates, and live birth rate between the two groups. Logistic regression analysis indicated that SDF was not a prognostic predictor of positive HCG, clinical pregnancy, miscarriage, and live birth. SDF was not associated with clinical outcomes either in ICSI or IVF cycles during vitrified-warmed single-blastocyst transfer cycles.

Intracytoplasmic Sperm Injection Cycle Outcomes in Women Aged 40 Years and Over

Objective: The main detrimental factor in female fertility is maternal age. Clinical pregnancy and live birth rates decline gradually with age and this decline is dramatical over 40 years. In this study, we examined the in vitro fertilization/intracytoplasmic sperm injection cycle outcomes of women aged 40 years and over. Study Design: This retrospective study included 336 fresh in vitro fertilization/ICSI cycles of women aged ≥40 years. Six groups, stratified by one-year intervals were composed according to age: 40 years; 41 years; 42 years; 43 years; 44 years, and ≥45 years. The primary outcomes were the clinical pregnancy and live birth rates. Results: The clinical pregnancy rate was 18.6% in 40 years old women and it decreased to 4% in women aged ≥45 years. The live birth rates in women aged 40 and 41 years (10% and 6.1% respectively) were higher than the live birth rates in women aged 42 and 43 years (4.3% and 3.8% respectively). There was no live birth in women aged 44 and over. The miscarriage rate was 46.2% at age 40 and it increased to 100% at age 44 and over. Conclusion: The clinical pregnancy and the live birth rates decreased significantly with every single year after 40 years of age. In patients between the ages of 40-43, the live birth rate is acceptable but starting an in vitro fertilization cycle beyond 43 years of age does not seem to be reasonable.

Influence of sperm morphology in ICSI cycle outcomes: a retrospective study

Aim: To determine whether sperm morphology has any influence on the assisted reproductive technique (ART) outcome in in vitro fertilization (IVF)-intracytoplasmic sperm injection (ICSI) cycles. Objectives: To assess whether sperm morphology affects IVF-ICSI outcome. Method: This study was a retrospective analysis of data from 1000 couples who had undergone ART cycle at Southend Fertility & IVF, Delhi between January 2016 and January 2021. The recruited patients were divided into two groups: Group A: Group A was the study group, and included 600 patients with an abnormal sperm morphology (morphology of <4%); Group B: Group B was the control group, and included 400 patients with a normal sperm morphology (morphology of >4%). The two groups were compared in terms of fertilization rate, embryo development rate, grade A embryo development rate, grade B embryo development rate, grade C embryo development rate, cleavage rate, embryo discarded or damage rate, and pregnancy rate. Results: Of the total 1000 ART-ICSI cycles analyzed the baseline characteristics, such as age, type of infertility, and semen parameters such as volume, count, motility, debris, agglutinations, grade A motility, grade B motility, total progressive (A+B) motility, morphology assessments, and type of protocol used for stimulation were comparable between the two groups. In the 1000 patients analyzed, a total of 6871 oocytes were injected at ICSI, of which 4049 oocytes belonged to group A and 2822 oocytes belonged to group B of the 6871 oocytes injected, 6275 got fertilized, giving a fertilization rate of 91.3%. On comparing the fertilization rate of the two groups, a fertilization rate of 90.1% (3650/4049) was observed in group A and 93.01% (2625/2822) was observed in group B, which was statistically different (P < 0.001). Of the various outcome parameters, a statistically significant difference was reported in the fertilization rate, cleavage rate, embryo development rate, grade A embryo development rate, and grade C embryo development rate in the two groups. No statistically significant difference was observed in the pregnancy rate. It was also observed that once fertilization occurred, embryo quality was good for all types of abnormal spermatozoa (grade A, good- to excellent-quality embryos), except for spermatozoa with broken necks, with which only good to excellent quality, or poor quality grade B and group C embryos were obtained. Conclusion: Sperm morphology influences the fertilization rate, cleavage rate, and the embryo development rate but there is no significant influence of sperm morphology on pregnancy rate.

Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies

BackgroundAzoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI.MethodsA retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group.ResultsCompared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05).ConclusionsNo negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

SUBTLE ENDOMETRIAL ABNORMALITIES DO NOT AFFECT THE PREGNANCY RATE IN FRESH ICSI CYCLES

The study aimed to determine whether the presence of subtle endometrial abnormalities discovered by office hysteroscopy (OH) affects the pregnancy rate in fresh intracytoplasmic sperm injection (ICSI) cycles. A prospective cohort study included women undergoing their first ICSI cycles was performed in assisted reproductive technology unit. Patients were divided into two groups: patients with subtle endometrial abnormalities diagnosed by OH (group 1, n=78), and patients with normal uterine cavity (group 2, n=122). OH was conducted post-menstrual and subtle abnormalities detected included hypervascularization, petechiae, mucosal elevation, micropolyps, pale endometrium and single adhesion band. The main outcome measures were clinical pregnancy and implantation rates The most common subtle abnormality observed was pale endometrium in 35.9% of cases. Both groups were comparable regarding the baseline clinical data and endometrial thickness. Additionally, total amount of gonadotrophins used, duration of stimulation, peak estradiol concentrations, peak endometrial thickness, number of mature follicles and number of embryos transferred were not significantly different between both groups. The implantation rate was similar in both groups (2.35 ± 0.83 vs. 2.22 ± 0.84, p=0.465). Also, the pregnancy rate was similar in both groups (38.5% vs. 33.6%, p=0.067) Subtle endometrial abnormalities diagnosed by OH do not negatively affect pregnancy and implantation outcomes in fresh ICSI cycles.

The outcomes of gonadotropins releasing hormone (GnRH) antagonist cycles with and without letrozole co-treatment in patients undergoing ICSI

Background: Letrozole is an aromatase inhibitor which has been proposed as a great tool in ovulation induction and infertility management. This study aimed to evaluate whether the use of letrozole in combination with gonadotropins and GnRH antagonist is superior to gonadotropins and GnRH antagonist alone in women undergoing ICSI treatment. Methods: This prospective randomized controlled trial was conducted during the period from October 2017 to March 2020 in a private fertility center. 112 participants were randomly allocated into two groups either intervention (Letrozole plus antagonist protocol) or control (antagonist protocol) groups. Participants were subjected to ovarian stimulation (OS) using a GnRH antagonist protocol. Only for the intervention group, letrozole was added from the second day of the cycle to the sixth day. On the day of hCG trigger, endometrial thickness, estradiol and progesterone level were assessed. The results were correlated to the outcomes of the Intracytoplasmic sperm injection (ICSI) cycle. Results: Both the intervention and control groups were balanced in respect to the demographic and clinical characteristics. The stimulation outcomes were comparable between the two studied groups. The clinical pregnancy rate and the ongoing pregnancy rate were comparable between both groups. There was no significant relation between the day of embryo transfer and either occurrence of clinical pregnancy or ongoing pregnancy among both groups' participants. Conclusions: Letrozole co-treatment in the first five days of gonadotropins stimulation in anatagonist cycles did not show any significant change in the pregnancy outcomes of ICSI cycles.

Semen parameters on the intracytoplasmic sperm injection day: Predictive values and cutoff thresholds of success

ObjectiveThis study was conducted to investigate the relationship of semen parameters in samples used for intracytoplasmic sperm injection (ICSI) with fertilization and pregnancy rates in infertile couples.MethodsIn this prospective study of Infertile couples with male factor infertility that had undergone ICSI, fractions of the same semen samples obtained for microinjection (to ensure the best predictability) were evaluated to determine the semen parameters and sperm DNA fragmentation index (DFI) on the day of oocyte recovery.ResultsIn total, 120 couples completed the study and were subdivided into fertilized (n=87) and non-fertilized couples (n=33). The fertilized couples were further classified into pregnant (n=48) and non-pregnant (n=39) couples. Compared to non-fertilized and non-pregnant couples, fertilized and pregnant couples showed statistically significantly higher sperm viability and percentage of normal sperm morphology, as well as significantly lower sperm DFI values. A receiver operating characteristic curve analysis of data from the 120 ICSI cycles showed that sperm viability, normal sperm morphology percentages, and sperm DFI were significant prognostic indicators of fertilization at cutoff values of 40%, 7%, and 46%, respectively. A sperm DFI of 46% showed sensitivity and specificity of 95% and 90%, respectively, for predicting fertilization, and no clinical pregnancies occurred in couples with a sperm DFI above 46%.ConclusionSemen parameters from the ICSI day sample, especially sperm viability, normal morphology, and DFI, had an impact on fertilization and pregnancy outcomes in ICSI cycles.