Intracytoplasmic Sperm Injection Procedure Research Articles

Melatonin levels and embryo quality in IVF patients with diminished ovarian reserve: a comparative study

BackgroundMelatonin, a hormone found in various bodily fluids and cells, is known for its potent antioxidative, anti-apoptotic, and endocrine regulatory properties. This study aimed to analyze melatonin levels in patients with diminished ovarian reserve (DOR) and its impact on embryo quality.MethodsWe enrolled 85 women who were undergoing in vitro fertilization or intracytoplasmic sperm injection procedures, including normal ovarian reserve (NOR, n = 27), pathological DOR (DOR-Path, n = 25), and physiological DOR (DOR-Phy, n = 33). Melatonin levels in patient serum and follicular fluid were assessed using ELISA, and correlations between melatonin levels and indicators of embryo quality were examined.ResultsOur findings indicate that melatonin levels in the follicular fluid and basal serum of the DOR-Path and DOR-Phy groups were lower compared to the NOR group (P < 0.05). However, no significant differences in melatonin levels were found between the DOR-Path and DOR-Phy groups (P > 0.05). Additionally, the concentration of melatonin in the follicular fluid of the NOR group was significantly higher than in their serum (P < 0.001). Lastly, a significant correlation was discovered between melatonin levels in serum and follicular fluid and parameters of ovarian reserve and embryonic development (P < 0.05).ConclusionsMelatonin levels in DOR patients may impact embryo quality, offering insights into potential DOR pathogenesis and opportunities to enhance treatment outcomes in these patients.

Effect of body mass index on ovarian reserve and ART outcomes in infertile women: a large retrospective study

BackgroundObesity poses a significant global health challenge, with profound implications for women’s reproductive health. The relationship between ovarian reserve and body mass index (BMI) remains a subject of debate. While obesity is generally associated with poorer outcomes in assisted reproductive technology (ART), the evidence remains inconclusive. This study aimed to investigate the effect of pre-pregnancy BMI on ovarian reserve and ART outcomes in infertile patients.MethodsWe conducted a retrospective cohort study involving women who underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) procedures at Tongji Hospital between 2016 and 2023. The study included 30,746 initial fresh cycles and 5,721 singleton deliveries. Patients were stratified by age and further categorized into four BMI groups: lean (< 18.5 kg/m²), normal weight (18.5–24.9 kg/m²), overweight (25.0–29.9 kg/m²), and obese (≥ 30.0 kg/m²). The primary endpoints of the study were pregnancy and perinatal outcomes. To explore the association between BMI and these outcomes, we adjusted for relevant confounding factors and utilized multivariate linear regression models, complemented by multifactorial logistic regression analyses.ResultsAnti-Müllerian hormone (AMH) levels were significantly lower in the overweight and obese groups compared to the normal weight group. After adjusting for age, a negative correlation was found between AMH and BMI in the age subgroups of 20–30 and 30–35 years. Among women aged 20–35 years, those in the overweight and obese groups had significantly fewer retrieved oocytes, mature oocytes, and two-pronuclear (2PN) embryos than their normal weight counterparts. Despite these differences, pregnancy outcomes in the overweight and obese groups were comparable to those in the normal weight group across all age categories. Additionally, obesity was linked to an increased risk of gestational diabetes mellitus, hypertensive disorders of pregnancy, and macrosomia.ConclusionsAn age-related decrease in AMH levels was evident with increasing BMI. Although being overweight or obese is associated with poorer embryo and perinatal outcomes, it does not seem to have a substantial impact on fertility.

Reproductive characteristics and success rate of intracytoplasmic sperm microinjection in spinal cord injury infertile men: Retrospective cohort study.

Thanks to modern methods of assisted reproduction (ART), parenthood has become an attainable goal for couples in which the male partner has experienced spinal cord injury (SCI). The aim of our study was to determine the success of the treatment of infertile patients with SCI with intracytoplasmic sperm injection (ICSI) of cryopreserved sperm obtained by the testicular sperm aspiration (TESA) procedure. In this retrospective study 156 infertile couples were included, in which the male partner is primarily infertile due to azoospermia. Infertile couples were divided into two groups. The first group (n= 82) includes men with SCI, and the second (n= 74) men with obstructive azoospermia (OA) as the cause of infertility. All infertile men were examined and processed in the diagnostic procedure, and based on the urological findings, surgical extraction of sperm from the testicles was indicated. Exclusion criteria were the age of women over 40 and men over 45. We found that the quality of sperm was worse in the group with SCI, compared to the group with OA, but without statistical significance. Zenica and Johnsen score (p= 0.001; p= 0.000) showed worse semen characteristics in the group with SCI. There were no significant differences in the average number of embryos (p= 0.698), pregnancy rates per cycle (p= 0.979) and pregnancy rates per embrio transfer (ET), clinical pregnancy rates per ET (p= 0.987) and delivery rates per ET (p= 0.804) in couples with SCI, compared to couples with OA. Based on the results of this research, the TESA and ICSI procedures can be recommended as a successful method in the treatment of male infertility caused by azoospermia due to SCI.

Live Birth after Cleavage-Stage versus Blastocyst-Stage Embryo Transfer in Assisted Reproductive Technology: A Randomised Controlled Study.

Blastocyst stage transfer appears to improve pregnancy outcomes. The aim of this study is to evaluate the pregnancy results between fresh cycle blastocyst stage embryo transfer and cleavage stage embryo transfer in patients who undergo intracytoplasmic sperm injection (ICSI). This randomised clinical trial study was conducted at the Infertility Research Centre of Milad Hospital in Mashhad, Iran from 2018 to 2020 on 240 infertile women who presented for their first ICSI procedure. These patients were assigned to receive either cleavage embryo transfer (n=112) or blastocyst stage transfer (n=107). Pregnancy outcomes were measured in both groups. There were no differences regarding age, body mass index (BMI), serum follicle-stimulating hormone (FSH), duration of infertility, and aetiology of infertility between the groups (P>0.05). There were more follicles, total oocytes, and metaphase II (M2) oocytes in the blastocyst stage group. Considerably more cleavage stage embryos were transferred compared to the number of transferred blastocysts (P=0.001). The blastocyst group had more vitrified embryos than the cleavage group (P=0.000). The rates of implantation (P=0.332), chemical pregnancy (P=0.165), clinical pregnancy (P=0.694), and live births (P=0.727) were higher in the blastocyst group, but they were not significantly different. The rate of abortion was also not significantly higher in the blastocyst group (P=0.296). Blastocysts transferred in the fresh cycle of an ICSI procedure may be more advantageous compared to cleavage stage embryo transfer (registration number: IRCT20181030041503N1).

P-135 Impact of TE-biopsy practitioners on PGT-A results and live-birth outcomes after single euploid blastocyst transfer: comparison of 19 TE-biopsy practitioners in a single center

Abstract Study question Can TE-biopsy practitioners impact pre-implantation genetic test for aneuploidy (PGT-A) results, pregnancy, and live-birth outcomes in a single center that has nineteen biopsy practitioners? Summary answer Yes. Trophectoderm (TE)-biopsy practitioners, especially micromanipulation experience, can impact PGT-A results but do not impact pregnancy or live-birth outcomes after single euploid blastocyst transfer (SeBT). What is known already Recently, PGT-A has become important in in vitro fertilization treatment. However, several factors can impact the accuracy and interpretation of PGT-A results. Several studies have suggested that TE-biopsy practitioners can impact PGT-A results. However, these findings are controversial because these previous studies used data from a low number of TE-biopsy practitioners and from multiple centers. There is, therefore, still limited knowledge of the impact of TE-biopsy practitioners on PGT-A results and their clinical outcomes. Study design, size, duration A total of 3,589 PGT-A cycles (average maternal age, 40.9±3.2; 1,168 patients; February 2020 to November 2022) were retrospectively analyzed. Furthermore, 779 single frozen euploid blastocyst transfer (SeBT) cycles were analyzed. First, we investigated the association between TE-biopsy practitioners and transferable blastocysts that included euploid and low-level mosaic blastocysts (LLM, 20 to 40% abnormal cells). Second, we analyzed the impact of TE-biopsy practitioners on pregnancy and live-birth outcomes after SeBT. Participants/materials, setting, methods The nineteen practitioners (Years of intracytoplasmic sperm injection (ICSI) experience: 2 to 20 years) who received the same training performed a total of 5,613 TE-biopsies for PGT-A. The same practitioner performed both the biopsy and the tubing procedures for each blastocyst they biopsied. The Wald test for multivariable logistic regression analysis (mLR) was used to determine the impact of TE-biopsy practitioners. The confounding factors were selected by univariable logistic regression analysis before mLR. Main results and the role of chance TE-biopsy was carried out by the flicking method with laser assistance and it was taken five to seven TE cells. Overall, 22.9% of blastocysts were transferable blastocysts (euploid: 21.1%, LLM: 1.8%). The results were inconclusive for 0.1% of the blastocysts. Maternal/paternal age, number of previous egg retrieval/embryo transfers, anti-Müllerian hormone, sperm characteristics, indication for PGT-A, number of previous deliveries/chemical abortions, blastocyst quality (the day of vitrification, inner diameters of blastocysts, and Gardner grading) were used as confounding factors on the mLR. TE-biopsy practitioners were significantly associated with both ensuring transferable blastocysts and LLM in transferable blastocysts (p &amp;lt; 0.05). Additionally, the practitioner characteristics that impact ensuring transferable blastocysts or LLM in transferable blastocysts were assessed in a sub-analysis. Years of ICSI experience (p &amp;lt; 0.05), number of ICSI performed during the last years (p &amp;lt; 0.05), and number of TE-biopsies at that point (p &amp;lt; 0.05) were associated with ensuring transferable blastocysts and LLM in transferable blastocysts. On the other hand, TE-biopsy practitioners did not have a significant impact on biological pregnancy, chemical abortion, pregnancy, or live-birth rates after SeBT. Limitations, reasons for caution This study did not determine the influence of TE biopsy itself and the laser assistance on PGT-A and clinical results. And this study was based on a minimal stimulation cycle. Moreover, the patient cohort in this study only included cases involving recurrent implantation failure or repeated pregnancy loss. Wider implications of the findings This study showed that TE-biopsy practitioners could impact decreasing transferable blastocysts, and this depends on their micromanipulation experience. This result suggests TE-biopsy training should be started when embryologists have gained several years of experience with ICSI procedures. This could prevent the decline in the efficacy of PGT-A for clinical results. Trial registration number not applicable

P-048 Morphokinetic analysis and implantation prediction of embryos derived from oocytes microinjected with Artificial Intelligence (AI) real-time assessed sperm

Abstract Study question Is embryo morphokinetic development affected by sperm quality assessed real time during Intracytoplasmic Sperm Injection (ICSI)? Summary answer Embryos derived from oocytes microinjected with low-quality sperm, analyzed by an AI-tool, exhibit a sluggish pace and a diminished potential for successful implantation. What is known already The study of embryo morphokinetics provides insights into the temporal aspects of cell division, which are pivotal for successful implantation. Key parameters such as time of division to 3 cells (t3), time to division to 5 cells (t5), time between division to 3 cells and subsequent division to 4 cells (s2) and duration of cell cycle two, i.e. time between division to 2 cells and division to 3 cells (cc2) are closely associated with implantation potential. Monitoring these markers allows clinicians to select embryos with optimal developmental trajectories, improving the chances of successful implantation and ultimately leading to healthier pregnancies. Study design, size, duration That is a single-centre and non-intervention study of 60 ICSI cycles. Seminal samples were assessed real-time with Sperm identification (SiD) software (v1.0, IVF2.0, LTD). Microinjected oocytes were maintained in culture in time-lapse incubators (EmbryoScope or EmbryoScope Plus). Injected sperm and their corresponding oocytes were individually followed-up to assess early embryo morphokinetics: t3, t5, s2, cc2 (273 total blastocysts) in a four-month period. Participants/materials, setting, methods Sixty ICSI cycles (35 from own oocytes and 25 from egg donation) were studied.418 sperm were assessed with SiD and classified as ‘low’, ‘medium’, ‘good’ or ‘best’, according to their motility parameters. Early embryonic morphokinetics were studied focusing on t3, t5, s2 and cc2 times. Ranges related with best implantation potential were studied according to previous works of our group: t3(34-40h), t5 (45-55h), cc2 (9-12h) and s2 (≤0,76h). Main results and the role of chance No discernible differences were observed in the parameters t3 and t5 among embryos within each sperm quality group. However, significant differences were identified in s2 and cc2 metrics between embryos of distinct sperm quality groups. Specifically, oocytes subjected to microinjection with ‘best’ or ‘good’ sperm exhibited an optimal average cc2 length (10.49 ± 5.12), in stark contrast to embryos derived from oocytes microinjected with ‘medium’ or ‘low’ sperm, which manifested an acelerated developmental pace (8.73 ± 5.55) (p &amp;lt; 0.05). Furthermore, embryos from the ‘best’ and ‘good’ groups displayed significantly shorter s2 lengths (1.16 ± 1.98 and 1.03 ± 1.62) compared to embryos from the ‘low’ group (2.73 ± 3.96). The latter exhibited an extended duration between the 3-cell division and subsequent division to 4 cells (p &amp;lt; 0.05). These findings lead us to infer that embryos arising from oocytes microinjected with ‘best’ or ‘good’ sperm possess superior implantation potential compared to those originating from oocytes microinjected with ‘medium’ or ‘low’ sperm, as indicated by SiD scores. The latter cohort appears to exhibit an alterated morphokinetic development and diminished implantation potential. Limitations, reasons for caution This project is limited by its retrospective and single-centre nature. In addition, studying later embryo morphokinetics parameters could provide a better description of embryo development and its relation with sperm quality assessed real-time. Finally, sperm morphology should also be included in SiD software because of its relation with reproductive success. Wider implications of the findings Selection of high-quality sperm to microinject into the oocytes, supported by real-time analysis, could contribute to improved morphological and kinetic characteristics of embryos, potentially increasing the chances of success in assisted reproductive procedures. In addition, it provides an opportunity to reduce subjectivity and inter-user variability of ICSI procedure. Trial registration number Not applicable

A brief history of technical developments in intracytoplasmic sperm injection (ICSI). Dedicated to the memory of J.M. Cummins.

Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technology for treatment of severe male infertility introduced into clinical practice in 1992. This review provides a brief history of the development of ICSI by acknowledging major developments in the field. The review addresses key developments in pre-clinical and early studies, how ICSI compares with in vitro fertilisation, long-term consequences, how the mechanistic approach to ICSI has changed in both manual and semi-automated approaches, and how sperm selection procedures are integrated into ICSI. From the beginnings using animal models in the 1960-1970s, the development of ICSI is a remarkable and transformative success story. Indeed, its broad use (70% of cycles globally) exceeds the need required for treating infertile males, and this remains a controversial issue. There remain questions around the long-term health impacts of ICSI. Furthermore, advances in automation of the ICSI procedure are occurring. An estimated 6million children have been born from the ICSI procedure. With further automation of sperm selection technologies, coupled with automation of the injection procedure, it is likely that the proportion of children born from ICSI will further increase.

The impact of SARS-CoV-2 mRNA vaccine on intracytoplasmic sperm injection outcomes at a fertility center in Iraq: A prospective cohort study.

Coronavirus disease 2019 (COVID-19) is a major public health problem that requires preventative vaccines. However, there is vaccine hesitancy among women of reproductive age in Iraq. This study aimed to investigate SARS-CoV-2 vaccination effects on intracytoplasmic sperm injection (ICSI) and related fertility parameters. The study population comprised 54 infertile patients undergoing the ICSI procedure at a fertility clinic: vaccinated (n = 17) and non-vaccinated (n = 37). SARS-CoV-2-IgG/mL was assayed in follicular fluid from patients. Fertility parameters were assessed using oocyte and embryo quality and pregnancy outcomes between study groups, with respect to the time interval from vaccination to ova pick up. There were no significant differences between non-vaccinated and vaccinated groups in respect of oocytes quality with regard to the mean number of picked up oocytes (p = 0.564), abnormal oocyte (p = 0.827), oocytes metaphase I and II (p = 0.306; p = 0.165), germinal vesicles (p = 0.076), grade I, II, and III fertilized oocytes (p > 0.05), and for maturation rate (p = 0.13). There were also no significant differences (p > 0.05) in embryo quality parameters with the mean number of grade I, II, and III fertilized oocytes and the fertilization rate, the number of transferred embryo (0.086). There were no significant differences between vaccinated and unvaccinated groups with respect to follicular fluid SARS-CoV-2-IgG (p = 0.854), and pregnancy outcomes (p = 0.550). The COVID-19 mRNA vaccine has no effect on ICSI, fertility parameters, and pregnancy outcome.

Influx of zwitterionic buffer after intracytoplasmic sperm injection (ICSI) membrane piercing alters the transcriptome of human oocytes.

Does piercing oocyte membranes during ICSI allow the influx of surrounding zwitterionic buffer into human oocytes and result in altered developmental competence? Human oocytes directed to IRB-approved research were used to determine the unrestricted influx of surrounding buffer into the oocyte after piercing of membranes via confocal fluorescence microscopy (n = 80 human MII oocytes) and the influence of the select buffer influx of HEPES, MOPS, and bicarbonate buffer on the oocyte transcriptome using ultra-low input RNA sequencing (n = 40 human MII oocytes). Piercing membranes of human MII oocytes during sham-ICSI resulted in the unrestricted influx of surrounding culture buffer into the oocyte that was beyond technician control. Transcriptome analysis revealed statistically significant decreased cytoskeletal transcripts in the pierced buffer cohorts, higher levels of embryo competency transcripts (IGF2 and G6PD) in the bicarbonate buffer cohort, higher levels of stress-induced transcriptional repressor transcripts (MAF1) in the HEPES and MOPS cohorts, and decreased levels of numerous chromosomal maintenance transcripts (SMC3) in the HEPES buffer cohort. The HEPES buffer cohort also revealed higher levels of transcripts suggesting increased oxidative (GPX1) and lysosomal stress (LAMP1). The influence of zwitterionic buffer on intrinsic cellular mechanisms provides numerous concerns for their use in IVF clinical applications. The primary concern is the ICSI procedure, in which the surrounding buffer is allowed influx into the oocytes after membrane piercing.Selecting a physiological bicarbonate buffer may reduce imposed stress on oocytes, resulting in improved embryo development and clinical results because intracellular MOPS, and especially HEPES, may negatively impact intrinsic biological mechanisms, as revealed by transcriptome changes. These findings further support the utilization of bicarbonate buffer as the oocyte-holding medium during ICSI.

Evaluation of Oxidative and Nitrosative Stress Markers Related To Inflammation in The Cumulus Cells and Follicular Fluid of Women Undergoing Intracytoplasmic Sperm Injection: A Prospective Study.

Oxidative/nitrosative stress in the oocyte microenvironment could have an impact on intracytoplasmic sperm injection (ICSI) outcomes. The presence of reactive oxygen species (ROS) and reactive nitrogen species (RNS) can stimulate the secretion of pro-inflammatory cytokines, leading to chronic inflammation and potentially affecting embryo as well as oocyte quality. This study aimed to examine the relationship of lipid peroxidation [measured by the malondialdehyde (MDA) assay] with protein carbonyl [measured by the 2,4 dinitrophenylhydrazine (DNPH) assay] levels in cumulus cells (CCs), as well as nitric oxide (NO), peroxynitrite (ONOO-), and C-reactive protein (CRP) levels in follicular fluid (FF). The potential relationship of these levels with ICSI outcome was also evaluated. In this prospective study, 63 FF samples and their corresponding CCs were collected for ICSI procedures. Spectrophotometry was used to assess levels of DNPH, MDA, NO, and ONOO-. CRP levels were evaluated using an immunoturbidimetric assay. The patients under 37 years with normal ovarian reserve had significantly lower levels of MDA, DNPH, NO, ONOO-, and CRP compared to those over 37 years. Additionally, we observed higher levels of MDA, DNPH, NO, ONOO-, and CRP in the group with an oocyte maturity rate of less than 60%. No significant difference was observed between the DNPH levels and factors such as infertility duration, embryo quality, pregnancy, or the number of retrieved oocytes. A higher level of MDA, NO, ONOO-, and CRP was found to be significantly related to the lower number of retrieved oocytes, longer periods of infertility, poor embryo quality, and negative pregnancy outcomes. Oxidative/nitrosative stress, linking to inflammation in the oocyte microenvironment, can be considered as a potentially useful biomarker for assessing the development and competence of oocytes and embryos and predicting ICSI outcomes.

Intuitive Cell Manipulation Microscope System with Haptic Device for Intracytoplasmic Sperm Injection Simplification.

In recent years, the demand for effective intracytoplasmic sperm injection (ICSI) for the treatment of male infertility has increased. The ICSI operation is complicated as it involves delicate organs and requires a high level of skill. Several cell manipulation systems that do not require such skills have been proposed; notably, several automated methods are available for cell rotation. However, these methods are unfeasible for the delicate ICSI medical procedure because of safety issues. Thus, this study proposes a microscopic system that enables intuitive micropipette manipulation using a haptic device that safely and efficiently performs the entire ICSI procedure. The proposed system switches between field-of-view expansion and three-dimensional image presentation to present images according to the operational stage. In addition, the system enables intuitive pipette manipulation using a haptic device. Experiments were conducted on microbeads instead of oocytes. The results confirmed that the time required for the experimental task was improved by 52.6%, and the injection error was improved by 75.3% compared to those observed in the conventional system.

Oocyte activation with phospholipase Cζ mRNA induces repetitive intracellular Ca2+ rises and improves the quality of pig embryos after intracytoplasmic sperm injection.

For the intracytoplasmic sperm injection (ICSI) procedure in pigs, an electrical pulse (EP) has been used as an effective method for oocyte stimulation, but unlike sperm, EP is unable to induce Ca2+ oscillations. In this study, we investigated the effects of generating artificial Ca2+ oscillations with phospholipase Cζ (PLCζ) mRNA, a candidate sperm factor, on fertilization, embryonic development, and gene expression after ICSI. Firstly, the concentration of PLCζ mRNA of a fixed volume (1.0 pl) that would induce a pattern of Ca2+ rise similar to that of in vitro fertilized (IVF) sperm was examined and determined to be 300 ng/μl. Secondly, the effects of oocyte stimulation methods on fertilization and embryonic development were investigated. ICSI-oocytes were activated by EP (EP group) or by PLCζ mRNA (PLCζ group). Furthermore, IVF-oocytes (IVF group) and ICSI-oocytes with and without an injection of buffer (buffer and untreated groups, respectively) were used as controls. It was found that the rates of normal fertilization in the PLCζ and EP groups were significantly higher than those in the buffer and untreated groups. The blastocyst formation rates did not differ among the groups. The embryo quality in the EP group was inferior to those in the PLCζ and IVF groups. Additionally, the expression level of a proapoptosis-related gene (Caspase-3) in the EP group was significantly higher than those in the PLCζ and IVF groups. Our data suggest that oocyte activation by PLCζ mRNA has the effect of improving embryo quality.

Evaluation of medical equipment of the infertility treatment centers in North western of Libya

Background Cell culture, particularly in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) are both of which assisted reproductive technology (ART) approaches that are most frequently utilized to treat infertility around the world. Objective Highlight the differences and similarities in instruments and equipment in ART laboratories that govern their success, limitations, and potential differences between laboratories by identifying the facilities of laboratories to improve outcomes of ICSI and IVF techniques in clinical centers on the western coast of Libya. Method: descriptive study, comparison of five (5) assisted reproduction laboratories instrument and equipment parameters using the same quality-control application in clinical centers on the western coast of Libya.Result: Among the seven clinical centers specializing in the diagnosis and treatment of infertility that were contacted, four infertility treatment centers and the Medical Research Center (MRC) provided a response. The MRC and four clinical centres had good laboratory facilities for ICSI and IVF techniques compared to Jordan ART laboratory facilities.Conclusion: This study highlights the imperative of enhancing laboratory infrastructure to support optimal implementation of ICSI and IVF procedures.

ICSI using testicular spermatozoa after failure of ICSI with ejaculated spermatozoa could be a good choice: A propensity score-matched cohort study.

Ejaculated spermatozoa are considered to possess a higher fertilisation potential than testicular spermatozoa. In selected cases, the use of testicular spermatozoa from non-azoospermic infertile men resulted in a higher implantation and pregnancy rate than the use of ejaculated spermatozoa. The primary objective was to compare the live birth rate and cumulative live birth rate between couples with failed intracytoplasmic sperm injection procedure using ejaculated spermatozoa who subsequently had an intracytoplasmic sperm injection cycle with testicular spermatozoa and those who subsequently had an intracytoplasmic sperm injection cycle with ejaculated spermatozoa. The secondary objective was to determine the indications for the use of testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa. A retrospective study of matched couples using propensity score matching analysis was performed. After an intracytoplasmic sperm injection failure (cycle_1), intracytoplasmic sperm injection with either ejaculated spermatozoa (ejaculated sperm group), or testicular spermatozoa (testicular sperm group), was performed (cycle_2). The matching was on intracytoplasmic sperm injection performed in cycle_1 according to spermatozoa used (testicular or ejaculated) in cycle_2. Logistic regression was used to evaluate the influence of sperm origin on cumulative live birth rate. Univariate analysis on parameters of cycle_1 was used to identify the prognostic factors to propose an intracytoplasmic sperm injection with testicular spermatozoa in case of cycle_1 failure. The study outcomes were live birth rate and cumulative live birth rate. Among the 6034 couples available, 63 were selected to constitute the testicular sperm group and 63 were selected by propensity score matching to constitute the ejaculated sperm group. After matching, the DNA fragmentation index was higher in the testicular sperm group (13.43%±9.65% vs. 8.93%±4.47%, p=0.013); no significant difference was observed for the fertilisation rate, the number of obtained embryos, blastulation rate and frozen embryo rate. In cycle_2, the live birth rate was higher in the testicular group (22.2% vs. 0.0%, p<0.001), as was the cumulative live birth rate (25.4% vs. 6.3%, p=0.065). The prognostic factors identified for the proposal of intracytoplasmic sperm injection procedure with testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa were: teratozoospermia, cryptozoospermia and high DNA fragmentation index. According to the present study and current knowledge, the use of testicular spermatozoa after failed intracytoplasmic sperm injection procedure in non-azoospermic men could be proposed instead of sperm donation in case of high sperm DNA fragmentation index, cryptozoospermia and teratozoospermia. A good oocyte response to ovarian stimulation during the previous assisted reproductive technology attempt will increase the chance of success. Although the main limitation of the current study is its retrospective nature, the use of the propensity score matching to perform causal inference study increases its reliability. The present study supports that the use of testicular spermatozoa outside the classical indication of azoospermia is a good option when the indication is well established. However, before proposing a testicular biopsy, an improvement in sperm characteristics should be considered by treating the causes of sperm alteration.

#166 : Investigating the Effect of Parental Age on Intracytoplasmic Sperm Injection Reproductive Outcomes in Indonesia: Prognostic Tools and Prediction Approaches to Maximize Human Embryo Implantation

Background and Aims: Reproductive issues impact one in eight Indonesian married couples. Intra Cytoplasmic Sperm Injection (ICSI) is still expensive; hence many couples don’t get evaluated or enroll fertility programs. We decided to study the correlation between age, hormonal status, and ICSI embryo quality because Indonesia’s infertility rate is rising. This is the first investigation into the factors behind ICSI outcomes in Indonesia. Method: In 2022, two fertility centers in Surabaya and Tangerang, Indonesia, performed ICSI on couples. 150 couples receiving ICSI at two research sites comprise the sample. Surgically retrieved or frozen sperm cycles and incomplete medical records were excluded. The Spearman test was used to investigate the correlation between parental age, hormonal status, and ICSI embryo quality on day 1 and day 3. (D1 and D3). Results: 150 ICSI cycles were studied. The hormonal baseline varied from normal to subnormal, while paternal and maternal age ± SDs were 37.03 ± 6.08 and 34.28 ± 5.11. The Spearman test demonstrated a significant negative connection between paternal age and embryo quality D1 (p&lt;0.005, r=-0.363) and D3 (good category: p&lt;0.005, r=-0.172; intermediate category: p&lt;0.005, r=-0.250; poor category: p&lt;0.005, r=-0.221). The Spearman test showed a significant negative connection between maternal age and embryo quality D1 (p&lt;0.005, r=-0.413) and D3 (good category: p&lt;0.005, r=-0.231; intermediate category: p&lt;0.005, r=-0.294; poor category: p&lt;0.005, r=-0.291). Conclusion: This study reveals that married couples have a substantial impact on their hormonal condition, the rate of fertilization, the quality of D1 and D3 embryos, and the number of embryos generated following ICSI. To increase the success rate of ICSI procedures, it is necessary to maximize fertility at a younger age.

Effects of histones related to sperm chromatin on embryo development and ART outcomes.

In the process of spermatogenesis and maturation, histones of the sperm nucleus were gradually replaced by protamine. Abnormal sperm nucleoprotein histotype conversion can make sperm DNA unstable and affect sperm function. The aim of this study is to investigate the impact of high and low proportion of sperm histone positivity in semen sample on embryonic development and assisted reproductive technology results, and to evaluate its diagnostic value in assisted reproduction. Sperm nuclear status was detected with aniline blue staining. Under acidic conditions, aniline blue combines with histones rich in lysine residues to form blue compounds. The groups were divided according to the critical value of sperm histone positive ratio of 30%. Using the intracytoplasmic sperm injection procedure, the fertilization rate and normal fertilization rate in the normal sperm histone positive ratio group were significantly higher than those in the abnormal group, and the difference was statistically significant (P < .001). Using the in vitro fertilization procedure, the effect of sperm histone positive ratio on each index was not statistically different. Overall the study provides some preliminary evidence that abnormal sperm histones may be a factor that affects the fertilization success of intracytoplasmic sperm injection procedures. However, more research is needed to confirm this finding to determine the exact mechanism by which abnormal sperm histones affect fertilization.

Ayurvedic management of male infertility due to severe oligospermia associated with hypotrophy of testes and hyperviscosity: A case report

Abstract: Infertility is defined as the failure of a couple to achieve conception within one or more years of regular unprotected coitus. Male infertility is any health issue in a man that lowers the chances of his fertile female partner getting pregnant. Males are found to be solely responsible for 20%–30% of infertility cases and contribute to 50% of overall cases. Testicular hypotrophy is a potential cause of reduced semen quality which results in decreased chances of successful fertilization of an ovum. In this case report, a 32-year-old male patient presented with complaints of primary infertility due to severe oligospermia associated with hypotrophy of testes and hyperviscosity since three years and six months of married life, with a 26-year-old female having regular menstruation. They underwent internal medication and Intrauterine Insemination (IUI) two times and were suggested an Intracytoplasmic Sperm Injection (ICSI) procedure. As per Ayurveda parameters, the case was diagnosed and medicines were administered in three phases: an initial phase of Deepana (~enhancing metabolic fire) and Anulomana (~proper elimination of flatus, feces, urine, etc.) Chikitsa (~treatment), followed by Granthibhoota shukradushti (~hyperviscous semen) Chikitsa in the second phase and Dhatu vriddhikara (~nourishment of the tissues) and Vajikarana chikitsa (~aphrodisiac treatment) in the last phase. After treatment, total sperm count increased from 1 million/cc to 20 million/cc, active motility improved from 3% to 10%, viscosity became normal, liquefaction time decreased from 1 h to 30 min and ultimately resulted in pregnancy followed by delivery of a healthy male baby weighing 2.75 kg.

Ensembling noisy segmentation masks of blurred sperm images

Background:Sperm tail morphology and motility have been demonstrated to be important factors in determining sperm quality for in vitro fertilization. However, many existing computer-aided sperm analysis systems leave the sperm tail out of the analysis, as detecting a few tail pixels is challenging. Moreover, some publicly available datasets for classifying morphological defects contain images limited only to the sperm head. This study focuses on the segmentation of full sperm, which consists of the head and tail parts, and appear alone and in groups. Methods:We re-purpose the Feature Pyramid Network to ensemble an input image with multiple masks from state-of-the-art segmentation algorithms using a scale-specific cross-attention module. We normalize homogeneous backgrounds for improved training. The low field depth of microscopes blurs the images, easily confusing human raters in discerning minuscule sperm from large backgrounds. We thus propose evaluation protocols for scoring segmentation models trained on imbalanced data and noisy ground truth. Results:The neural ensembling of noisy segmentation masks outperforms all single, state-of-the-art segmentation algorithms in full sperm segmentation. Human raters agree more on the head than tail masks. The algorithms also segment the head better than the tail. Conclusions:The extensive evaluation of state-of-the-art segmentation algorithms shows that full sperm segmentation is challenging. We release the SegSperm dataset of images from Intracytoplasmic Sperm Injection procedures to spur further progress on full sperm segmentation with noisy and imbalanced ground truth. The dataset is publicly available at https://doi.org/10.34808/6wm7-1159.

Human oocytes image classification method based on deep neural networks

BackgroundThe effectiveness of in vitro fertilization depends on the assessment and selection of oocytes and embryos with the highest developmental potential. One of the tasks in the ICSI (intracytoplasmic sperm injection) procedure is the classification of oocytes according to the stages of their meiotic maturity. Oocytes classification traditionally is done manually during their observation under the light microscope. The paper is part of the bigger task, the development of the system for optimal oocyte and embryos selection. In the hereby work, we present the method for the automatic classification of oocytes based on their images, that employs DNN algorithms.ResultsFor the purpose of oocyte class determination, two structures based on deep neural networks were applied. DeepLabV3Plus was responsible for the analysis of oocyte images in order to extract specific regions of oocyte images. Then extracted components were transferred to the network, inspired by the SqueezeNet architecture, for the purpose of oocyte type classification. The structure of this network was refined by a genetic algorithm in order to improve generalization abilities as well as reduce the network’s FLOPs thus minimizing inference time. As a result, overline{Acc} at the level of 0.964 was obtained at the level of the validation set and 0.957 at the level of the test set. Generated neural networks as well as code that allows running the processing pipe were made publicly available.ConclusionsIn this paper, the complete pipeline was proposed that is able to automatically classify human oocytes into three classes MI, MII, and PI based on the oocytes’ microscopic image.

Obesity and fertility: a literature review of the association between parental obesity at the time of conception and embryo quality

Obesity [body mass index (BMI) ≥30 kg/m2] has been shown to be a risk factor in reducing the rate of successful in vitro fertilization (IVF) outcomes. In this systematized literature review, we investigate whether embryo quality specifically is affected by single-parent obesity (maternal or paternal) and obesity in both parents at the time of conception. We systematically searched for studies published over the last 10 years that graded embryo quality in conventional IVF and intracytoplasmic sperm injection procedures. Studies that did not use measured weight and height for BMI calculation were excluded. Ten eligible articles were found across 13 public databases. Comparing the results of 1845 couples, both female and male obesity independently caused faster preimplantation embryo development (P &lt;0.05). The fertilization rate was not affected by either female, male, or combined parental obesity (P&gt;0.05). Day 3 embryo quality does not seem to be affected by either female or male obesity alone. The 2 studies that investigated the effect of combined parental obesity on embryo quality reached contradictory findings. Male, but not female, &gt;7% BMI reduction following bariatric surgery significantly increased the number of top-quality embryos compared with their results before surgery (P&lt;0.05). We conclude that embryos of couples with single-parent obesity are not of lower quality compared with those of normal-weight parents. Combined parental obesity may affect embryo quality. BMI reduction could lead toward a successful outcome in obese infertile couples, regardless of whether they are undergoing IVF or not.