Intracytoplasmic Sperm Injection Technique Research Articles

P-776 School-age outcomes among IVF and ICSI -conceived children: a causal inference analysis using linked population-wide data

Abstract Study question What is the causal effect of method of oocyte insemination; standard in-vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI), on childhood development at school entry? Summary answer The developmental outcomes at school entry for children conceived by ICSI, without severe male factor infertility, are equivalent to those conceived via standard IVF. What is known already Over 12 million children have been conceived globally with the assistance of assisted reproductive technology (ART). Within ART, ICSI continues to increase as the most common mode of oocyte insemination, even when not clearly indicated (i.e. without male factor infertility). Large cohort studies have suggested that ART, and ICSI in particular, may be associated with an increase in congenital abnormalities, autism spectrum disorder, developmental delay and intellectual disability. However, the studies are heterogenous and the evidence conflicting. The association between the ICSI technique itself and long-term childhood development remains unclear. Study design, size, duration Causal inference methods were used to analyse observational data in a way that emulates a target randomised clinical trial. For example, patients where standard IVF was not an option, (e.g., due to severe male factor infertility), were excluded. The state-wide cohort included linked maternal and childhood administrative data from Victoria, Australia. To eliminate potential confounding associated with IVF, comparison was made with children conceived via standard IVF (rather than conception without assistance). Participants/materials, setting, methods The study included singleton infants conceived via ART and born between 2005-2013. The primary exposure was conception via ICSI, with those born following conventional IVF insemination as controls. The outcome examined was the Australian Early Development Census (AEDC), a broad assessment of development and well-being performed in Australian schools, every triennium, at school entry (age 4-6). We combined multiple imputation and inverse probability weighting with regression adjustment to estimate population average causal effects. Main results and the role of chance The final cohort included 4,035 IVF and ICSI conceived children across the two outcome cohorts. Linked records were available for 2,468 ICSI children and 1,567 IVF-conceived controls. Maternal characteristics between the two treatment cohorts were similar for age, parity, education, language background, and number of eggs collected during treatment cycle. The ICSI conceived children were generally from residential areas of lower socio-economic status and conception was more likely to involve donated gametes. The unadjusted rate of the primary outcome; AEDC-defined “developmental vulnerability”, was 10.83% among the IVF group and 10.14% among the ICSI group. Given the use of observational data, there were missing data and inherent differences in the covariate profile between exposure cohorts. Multiple imputation, bootstrapping and doubly robust inverse probability weighted regression adjustment modelling was utilised to allow a causal interpretation of results. There was no evidence of a causal effect of ICSI-conception on the risk of AEDC-defined developmental vulnerability at school entry compared with spontaneously conceived children; adjusted risk difference -0.8% (95% CI -2.7% to 1.1%) and adjusted risk ratio 0.92 (95% CI 0.76 to 1.12). The adjustment variable set was determined a priori via a modified Delphi procedure. Limitations, reasons for caution Children who did not attend school due to severe disability were excluded, leading to potential selection bias. Within our study cohort of 2,468 ICSI children, no increased risk of developmental vulnerability was identified. It is possible that unmeasured confounders could have led to bias in estimating the treatment effects. Wider implications of the findings This study, in contrast to previous evidence, suggests that empiric use of ICSI compared with conventional IVF insemination does not affect early childhood developmental outcomes. These findings provide important reassurance for current and prospective parents, and clinicians alike. Trial registration number not applicable

Monitoring Blood Glucose in Mouse Offspring after Intracytoplasmic sperm injection - Embryo Transfer.

Human lifespan is considerably long, while mouse models can simulate the entire human lifespan in a relatively short period, with one year of mouse life roughly equivalent to 40 human years. Intracytoplasmic sperm injection (ICSI) is a commonly used assisted reproductive technology in clinical practice. However, given its relatively recent emergence about 30 years ago, the long-term effects of this technique on human development remain unclear. In this study, we established the ICSI combined with embryo transfer (ET) method using a mouse model. The results demonstrated that normal mouse sperm, after undergoing in vitro culture and subsequent ICSI, exhibited a fertilization rate of 89.57% and a two-cell rate of 87.38%. Following ET, the birth rate of offspring was approximately 42.50%. Furthermore, as the mice aged, fluctuations in glucose metabolism levels were observed, which may be associated with the application of the ICSI technique. These findings signify that the mouse ICSI-ET technique provides a valuable platform for evaluating the impact of sperm abnormalities on embryo development and their long-term effects on offspring health, particularly concerning glucose metabolism. This study provides important insights for further research on the potential effects of the ICSI technique on human development, emphasizing the necessity for in-depth investigation into the long-term implications of this technology.

Effect of chemical activators after intracytoplasmic sperm injection (ICSI) on embryo development in alpacas

Low motility and low sperm concentration are characteristics of alpaca semen. Thus, the intracytoplasmic sperm injection (ICSI) technique represents an alternative to improve the reproductive capacity of the male. However, the effect of post-ICSI activation in alpaca is not yet known. The aim of the present study was to compare the effect of chemical activators on alpaca embryo development after ICSI. Alpaca ovaries were collected from a local slaughterhouse and transported to the laboratory. Category I, II and III oocytes were matured for 30 h at 38.5 °C. After ICSI, injected oocytes were randomly divided and activated as follows: i) 5 μM ionomycin for 5 min, ii) 7% ethanol for 4 min, iii) 5 μM ionomycin for 5 min, window period 3 h plus 7% ethanol for 4 min, iv) 5 μM ionomycin for 5 min, window period 3 h, a second ionomycin treatment for 5 min, followed by 1.9 mM 6-DMAP for 3 h, v) 10 mM SrCl2 for 3 h. Culture was carried out for 5 days in SOFaa at 38.5 °C. The cleavage rate was the lowest in the SrCl2 group, morula development was the lowest in the SrCl2 and without activation groups, and blastocyst stage was not different between groups (P<0.05). The rates with SrCl2 were lower in total embryos produced, whereas in transferable embryos they were lower with 2Io/6-DMAP and with SrCl2 (P<0.05). In conclusion, alpaca oocyte activation is more efficient with ionomycin and ethanol to produce transferable embryos.

Microdissection testicular sperm extraction in non-obstructive azoospermic patients with solitary testis: a retrospective case-control study.

Obtaining sperm from the testis surgically and using these sperm with the intracytoplasmic sperm injection technique, has opened the way for the possibility of biological fathering in men with non-obstructive azoospermia (NOA). We aimed to evaluate our sperm retrieval rate (SRR) by microdissection testicular sperm extraction (micro-TESE) in NOA patients with solitary testis. In this retrospective case-control study, fortyfive patients with NOA who had a congenital or acquired solitary testis were included, between September 2003 and January 2022. These patients were randomly matched with patients with NOA who had bilateral testes, using a 1:3 matching ratio. We found that SRR by micro-TESE in patients with solitary testis was similar to NOA patients with bilateral testis (51.1% vs. 50.4%). Age, infertility period, ejaculate volume, serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone, history of varicocelectomy, history of orchiopexy, testicular stimulation therapy before micro-TESE, testicular volume, genetic status, TESE side, micro-TESE success, complications and histopathological evaluation results of both groups were evaluated, there was a statistically significant difference in only serum FSH and LH levels. There was no difference between the groups in terms of complications and hormonal effects in the early postoperative period. Micro-TESE in NOA patients with solitary testis has similar sperm retrieval and complication rates as NOA patients with bilateral testis.

Bovine oocyte activation with bull or human sperm by conventional ICSI and Piezo-ICSI: Its relationship with PLCɀ activity.

The intracytoplasmic sperm injection (ICSI) technique has low efficiency in cattle. This has mainly been attributed to the oocyte activation failure due to oocyte and/or sperm factors. Our aim was to evaluate the effect of conventional ICSI and Piezo-ICSI with bull or human sperm on bovine oocyte activation and embryo development and to assess its relationship with the phospholipase C zeta (PLCɀ) activity of both species. In vitro matured bovine oocytes were randomly divided into five groups and were fertilized as follows: conventional ICSI using bovine sperm with chemical activation (control), conventional ICSI using bovine sperm, Piezo-ICSI using bovine sperm, conventional ICSI using human sperm, and Piezo-ICSI using human sperm. PLCɀ activity was determined in bull and human sperm samples. Within the groups using bull sperm, the oocytes fertilized by conventional ICSI had the lowest values of 2 pronuclei (PN) formation and cleavage, Piezo-ICSI increased both percentages and ICSI + chemical activation presented the highest 2 PN, cleavage, and blastocyst rates (p < 0.05). Within the groups using human sperm, the oocytes fertilized by Piezo-ICSI presented higher 2 PN and cleavage rates than those activated by conventional ICSI (p < 0.05). Piezo-ICSI with human sperm increased bovine oocyte activation as much as conventional ICSI + chemical activation with bovine sperm (p < 0.05). Higher values of PLCɀ activity were found in human sperm compared with bovine sperm (p < 0.05). Our results suggest that the higher stability of the bovine sperm in combination with its relatively low content of PLCɀ impairs bovine oocyte activation after ICSI.

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.

The birth of a healthy child in a married couple with a history of multiple unsuccessful attempts at ART by selecting sperm on oocyte-cumulus complexes. Case report

In male infertility, couples must use assisted reproductive technologies with the intracytoplasmic sperm injection (ICSI) technique. A significant disadvantage of ICSI is bypassing natural selection barriers since selective processes in the female reproductive tract are replaced by the choice of an embryologist based on sperm motility and morphological criteria. Various selection methods are being actively studied, bringing fertilization by ICSI closer to physiological, ensuring the selection of sperm with high DNA quality and good potential for fertilization. The article presents a clinical case of the birth of a healthy child in a married couple with severe male infertility and many ineffective attempts at assisted reproductive technologies using the method of sperm selection on oocyte-cumulus complexes. The child is physically healthy and develops according to age.

Oligoasthenozoospermic and Normozoospermic Indian Males Yielded Similar Fertilization Capability by Intracytoplasmic Sperm Injection

Background: The present study evaluated the fertilization outcomes by intra-cytoplasmic sperm injection (ICSI) technique with oligoastheno-zoospermic (OAZ) compared to normozoospermic (NZ) semen. Purpose: To find the fertilizing capability of human sperm in oligo-asthenozoospermic and normozoospermic semen samples. Methods: Fresh semen ejaculates belonging to NZ and OAZ groups (n=50 in each group) patients attending the infertility clinics were evalu-ated for semen volume, sperm motility, and sperm concentration. Ferti-lization rates with ICSI along with post-capacitation motility and con-centration were studied in both groups. Results: The motility and concentration in fresh semen samples patients of the NZ group (32.44±1.50%, 71.96±6.45 M/ml, respectively) were significantly (P&lt;0.05) higher compared to the OAZ group (26.76±1.38, 10.31±0.48 M/ml, respectively).However, the results of our study re-vealed similar fertilization rates in-vitro by ICSI in the NZ (98%) and OAZ (98%) groups. Conclusion:The in-vitro processing by the ICSI technique is an effective approach to achieving good fertilization rates in sub-fertile males.

Characteristics of oocytes that failed to fertilize after intracytoplasmic sperm injection.

We assessed the morphologies of meiotic spindles in oocytes that failed to fertilize following intracytoplasmic sperm injection (ICSI) and identified factors contributing to failed fertilization. A total of 225 unfertilized oocytes were collected after ICSI. Oocytes were fixed and stained for tubulin and chromosomes. Meiotic spindle morphologies, chromosome alignment, and sperm nuclear decondensation were assessed to identify contributing factors to fertilization failure. We identified relationships between several factors and both abnormal spindle morphologies and sperm nuclear decondensation in oocytes that failed to fertilize. Three causes for unfertilized oocytes after ICSI were identified: (I) the absence of a sperm nucleus in the ooplasm; (II) failed oocyte activation; and (III) defects in pronucleus formation or migration. The rate of disarranged polar spindles in oocytes collected from women older than 35 years (73.3%; 33/45 oocytes) was significantly higher than that of those collected from women 35 years and younger (50.4%; 68/135 oocytes; odds ratio [OR]: 2.71, 95% confidence interval [CI]: 1.29-5.69, p = 0.009). The proportion of unfertilized oocytes with abnormal spindles and chromosome misalignment was significantly higher in oocytes collected from women older than 35 years than those from women 35 years and younger (62.2% vs. 41.5%, p = 0.016). The proportion of partially decondensed chromatin in the abnormal sperm morphology group was significantly higher than in the normal sperm morphology group (66.7% versus 52.9%, OR: 1.78, 95% CI: 1.01-3.11, p = 0.044). The main contributor to the failure of oocytes to fertilize after ICSI is failed oocyte activation. The ICSI technique used, the maternal age, and sperm morphology are also contributing factors in fertilization failure after ICSI.

What Does Intracytoplasmic Sperm Injection Change in Embryonic Development? The Spermatozoon Contribution.

The intracytoplasmic sperm injection (ICSI) technique was invented to solve severe male infertility due to altered sperm parameters. Nowadays, it is applied worldwide for the treatment of couple infertility. ICSI is performed with any available spermatozoon from surgery or ejaculated samples, whatever are the sperm motility, morphology or quantity. The aim of the present review was to study if embryo development and kinetics would be modified by (1) ICSI under the technical aspects, (2) the micro-injected spermatozoa in connection with male infertility. From published data, it can be seen that ICSI anticipates the zygote kinetics Furthermore, because fertilization rate is higher in ICSI compared to conventional in vitro fertilization (IVF), more blastocysts are obtained for clinical use in ICSI. Sperm and spermatozoa characteristics, such as sperm parameters, morphology and vitality, DNA content (levels of sperm DNA fragmentation, microdeletions, and chromosomal abnormalities), RNA content, epigenetics, and sperm recovery site (testicular, epididymis, and ejaculated), have an impact on fertilization and blastocyst rates and embryo kinetics in different ways. Even though ICSI is the most common solution to solve couples' infertility, the causes of male infertility are crucial in building a competent spermatozoa that will contribute to normal embryonic development and healthy offspring.

Cryptozoospermia: Should we use ejaculated sperm or surgically retrieved sperm for assisted reproductive technology?

In cryptozoospermic subjects, it may often may be difficult to secure motile sperm for assisted reproductive technology (ART). We examined the results of ART with frozen thawed ejaculated sperm in cryptozoospermic subjects and evaluated whether sperm retrieval surgery is necessary for such patients in our clinic. Between 2013 and 2021, we evaluated 197 cryptozoospermic patients. Age, endocrine panel at the time of the initial semen analysis, and anti-müllerian hormone levels at the time of the spouse's first egg retrieval were examined. Cryopreservation of ejaculated motile sperm collected essentially weekly over a 3-month period was carried out. ART data recorded was the number of egg retrieval cycles, normal fertilization rate, and clinical pregnancy rate. ART using frozen sperm as well as sperm ejaculated on the day of egg retrieval was possible in all cases. The normal fertilization rate was 70.4%, the clinical pregnancy rate per embryo transferred was achieved in 31.5% (870 cycles), and the live birth rate per case was 73.8%. Intracytoplasmic sperm injection (ICSI) was possible without sperm retrieval surgery in cryptozoospermia, resulting in 73.8% of live births per patient. Sperm identification, sperm processing, and ICSI technique are especially important in cryptozoospermia. Sperm retrieval surgery can be avoided in cryptozoospermic patients.

SPERM FACTORS AND EGG ACTIVATION: ICSI and the discovery of the sperm factor and PLCZ1.

The discovery of PLCZ1 nearly 20 years ago as the primary Ca2+ oscillation-inducing factor in the sperm of mammals represented a significant breakthrough in our quest to elucidate the molecules and pathways that promote egg activation during fertilization. The advent of the intracytoplasmic sperm injection (ICSI) technique, which made fertilization possible without sperm capacitation, acrosome reaction, and gamete fusion, strengthened the research that led to the discovery of PLCZ1 and became an essential clinical tool for humans. The use of ICSI combined with the detection of PLCZ1 expression and mutations in infertile patients established the fundamental role of PLCZ1 in human fertility while leading to the discovery of novel components of the perinuclear theca, the site of the residence of PLCZ1 in sperm before fertilization. Remarkably, the more extensive use of ICSI in species other than humans and mice revealed poor success and exposed gaps in our understanding of PLCZ1 release and/or activation. Similarly, fertilization using sperm from mouse models lacking Plcz1 has produced striking results whose true implications are yet to be determined. Nevertheless, answers to these unresolved questions will produce a complete picture of the adaptations and molecular players that mammalian species employ to ensure the success of the triggering event of embryo development that has linked generations since the beginning of times.

P-050 Reproductive results in vasectomized patients according to the time of the surgical procedure

Abstract Study question Is there a difference in reproductive outcomes in vasectomized patients related to the time of the surgical procedure undergoing ART? Summary answer The time-interval post-vascetomy sperm recovery not affected until 19 years after procedure, although p&amp;lt;0,05 in the men age and AR with &amp;gt; 20 years procedure What is known already Assisted reproductive techniques (ART) have helped vasectomized men who want to have children again, situations that are currently seen relatively frequently. However, an adequate period of time from surgery to initiate ART has not been established. For this reason, it is intended to retrospectively analyze the reproductive results with ICSI, in couples where the man has undergone a previous vasectomy, relating them to the time elapsed since the surgical procedure. Study design, size, duration Retrospective study from 2000 to 2019 of ICSI cycles with 200 patients with their own oocytes, 38 patients with donated oocytes, carried out at IVI Alicante and Murcia. The groups were distributed according to the time elapsed since the man was vasectomized and the treatment carried out: Group A (&amp;lt;10 years), B (11-19 years) and C (&amp;gt; 20 years) discriminated as follows: Own/donated oocytes: group A: 90/26, B: 70/9 and C: 40/3. Participants/materials, setting, methods Inclusion criteria: A total of 239 patients and their partners who undergo the intracytoplasmic sperm injection technique with their own and donated oocytes, before Testicular Sperm Extraction (TESE) surgery and freezing-thamwing a biopsy of testicular tissue after. The statistical analysis was carried out with the statistical software R, version 4.0, linear and multiple regression, establishing significant differences when p &amp;lt; 0.05. Main results and the role of chance The age of the woman was not significant for any of the time intervals (34,5, 35,5 and 35,6 years in own oocytes cycles and 40,6, 41,8 and 43,7 years in donated oocytes cycles), while a significant increase in the age of the man is observed as the obstructive interval increases in own oocytes cycles (42,5, 46,5 and 52,9 years with p &amp;lt; 0,05). The results in donated eggs cycles are 47,6, 50,9, and 42,7 years. Regarding reproductive results, the fertilization rate was not significant for any type of treatment.Pregnancy and implantation rates were higher in the period between 11 and 19 years, 40% (p &amp;lt; 0.05) / 30.1% in own oocytes cycles and donated oocytes cycles were 55.6 / 42.9%. A live newborn rate is reported in own oocytes cycles in group A: 28.9, group B: 37.1 and group C:25%, while in donated oocytes cycles the results are: Group A: 50, group B: 66.7 and group C: 0%. The abortion rates of groups A, B and C in own oocytes cycles are: 0, 14.2 and 33.3 (p &amp;lt; 0.05), and donated oocytes cycles are: 7,6, 0 and 0% respectively with p = 0,78. Limitations, reasons for caution More studies are necessary to get to affirm these statements with greater scientific rigor. Wider implications of the findings According to our studies, the reproductive results of those patients who have undergone post-vasectomy ART are not affected according to the time interval up to 19 years. From the age of 20, there is a trend in increasing results based on pregnancy and abortion. Trial registration number Not applicable for non clinical-trials

Microfluidic chips as a method for sperm selection improve fertilization rate in couples with fertilization failure.

Sperm quality plays a vital role in successful fertilization and pregnancy. Patients with fertilization failure (total failure or low-fertilization rate) despite having normal semen parameters are a challenging group whose sperm cannot fertilize the oocyte via the intracytoplasmic sperm injection (ICSI) technique. Microfluidics is offered as a new method for proper sperm sorting. This study aimed to evaluate sperm parameters, DNA fragmentation index (DFI), expression of phospholipase C zeta 1 (PLCZ1), and transition nuclear proteins 1 (TNP1) mRNAs in sperm selected by microfluidic sperm sorting (MSS) chip compared with conventional density gradient centrifugation technique in patients with fertilization failure following ICSI. Subsequence fertilization rate and embryo quality were assayed. Normal morphology and total motility were significantly higher, and DFI was significantly lower in sperm selected by the MSS chip in fertilization failure and control groups. The RT-PCR results demonstrated a significant increase in the expression of PLCZ1 and TNP1 genes in sperm of both groups selected by MSS chips compared to the DGC method. In addition, with the selected sperm by MSS chip, an increase in fertilization rate and improvement of embryo quality was obtained. The present study findings show that sperm sorting by the microfluidic method improves fertilization rate in patients with poor fertilization outcomes following ICSI.

Cumulus cell antioxidant system is modulated by patients' clinical characteristics and correlates with embryo development.

To study whether the cumulus cell antioxidant system varies accordingly to patients clinical characteristics' as age, infertility diagnosis, BMI, and stimulation protocol applied and if the antioxidant profile of cumulus cells could be used as a predictor of embryo development. A prospective study including 383 human cumulus samples provided by 191 female patients undergoing intracytoplasmic sperm injection during in vitro fertilization treatments from a local in vitro fertilization center and processed in university laboratories. Catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione S-transferase (GST) enzyme activity levels and reduced glutathione (GSH) levels were measured in cumulus oophorus cells individually collected from each aspirated cumulus-oocyte complex, and the results of each sample were compared considering the oocytes outcome after ICSI and patients clinical characteristics. A total of 223 other human cumulus samples from previous studies were submitted to a gene expression meta-analysis. The antioxidant system changes dramatically depending on patients' age, infertility diagnosis, stimulation protocol applied, and oocyte quality. SOD activity in cumulus cells revealed to be predictive of top-quality blastocysts for young patients with male factor infertility (P < 0.05), while GST levels were shown to be extremely influenced by infertility cause (P < 0.0001) and stimulation protocol applied (P < 0.05), but nonetheless, it can be used as a complementary tool for top-quality blastocyst prediction in patients submitted to intracytoplasmic sperm injection technique (ICSI) by male factor infertility (P < 0.05). Through a simple and non-invasive analysis, the evaluation of redox enzymes in cumulus cells could be used to predict embryo development, in a personalized matter in specific patient groups, indicating top-quality oocytes and improving success rates in in vitro fertilization treatments. The trial was registered at UFRGS Research Ethics Committee and Plataforma Brasil under approval number 68081017.2.0000.5347 in June 6, 2019.

Fertility and Sterility top videos of 2020

To objectively grade all video publications in Fertility and Sterility during 2020 and compile a list of the top 5 surgical videos. Descriptive presentation of the 5 highest scoring video publications from Fertility and Sterility in 2020 SETTING: Not applicable. Not applicable. J.F., Z.K., J.P.P., and S.R.L. acted as independent reviewers of all video publications. A standardized scoring method was used to score all videos. Up to 5 points were awarded for each of the following categories: scientific merit or clinical relevance of the topic, the clarity of the video, the use of innovative surgical technique, and video editing or the use of marking tools on the video to highlight the important features or surgical landmarks. This allowed a maximum score of 20 for each video. The numbers of YouTube views and likes were used as a tiebreaker if ≥2 videos scored similarly. The interclass coefficient from a 2-way random-effects model was calculated to assess for agreement between the 4 independent reviewers. A total of 20 videos were published in Fertility and Sterility during 2020. After averaging scores from all 4 reviewers, a list of top 5 videos was created (Table1). The overall interclass coefficient for the 4 reviews was 0.77 (95% confidence interval, 0.56-0.89). An overall substantial agreement was noted among the 4 reviewers. Five videos particularly stood out within the competitive peer reviewed publications. The subject matter of these videos ranged from complex surgical procedures, including transvaginal natural orifice surgery and advanced endometriosis to basic embryology laboratory topics that covered trophectoderm biopsy and novel intracytoplasmic sperm injection techniques.

COMPARISON OF MICROFLUIDIC SPERM SORTING (MFSS) VERSUS PHYSIOLOGICAL INTRACYTOPLASMIC SPERM INJECTION (PICSI) VERSUS DENSITY GRADIENT IN HIGH DNA FRAGMENTATION INDEX SPERM SAMPLES

To evaluate the best method for sperm separation in high DNA fragmentation index (DFI) sperm samples. A prospective randomised study was conducted from 1st January 2019 to 1st January 2021. Three hundred patients were randomised by computer generated list and divided into 3 groups. Group A (n=100) , in which sperm were processed by microfluidic sperm sorter (MFSS) while in group B (n=100), sperm were selected by Physiological Intracytoplasmic Sperm Injection (PICSI) technique and Group C (n=100), where sperm were processed by density gradient technique. Morphologically normal motile sperm were injected by Intracytoplasmic sperm injection (ICSI) technique in mature oocytes after all three techniques. The study period included all normozoospermic patients with high DNA fragmentation index (>25% ) while oligospermic, asthenozoospermic samples, patients with poor ovarian reserve and advanced age were excluded from the study. All A grade embryos were vitrified and transferred in frozen embryo replacement cycle. All the 3 groups were compared on the basis of fertilisation rate, day 3 grade A embryo development rate , clinical pregnancy rate and miscarriage rate. Cycle characteristics (female age, length of stimulation, gonadotrophin dose, number of oocytes and number of transferred embryos) were similar among all 3 groups. Between the 3 groups, there was a statistically significant increase observed in Group A (Microfluidic Sperm Sorting) in the day 3 grade A embryo development rate (60% vs. 42% vs. 38%, p-0.016) and clinical pregnancy rate (62% vs. 46% vs. 41%, p-0.049), while no statistical significant difference observed in fertilisation rate (82% vs. 78% vs. 76%, p-0.80) comparing with group B and group C. But significantly higher miscarriage rate ( 12% vs. 11% vs. 25%, p-0.019) observed in density gradient (Group C) technique. We have demonstrated that sperm sorted by microfluidic is not only correlated with better DNA integrity but also with the reproductive outcome. Using it in routine practice can help in reducing the extraneous affect of sperm processing techniques and achieving higher pregnancy rate and sustaining it comparing with PICSI and Density Gradient techniques.

P–084 Microfluidic Sperm Sorting (MFSS) technique versus Physiological Intracytoplasmic Sperm Injection (PICSI) technique in high DNA fragmentation index sperm samples

Abstract Study question To evaluate the effectiveness of using Microfluidic Sperm Sorting (MFSS) technique and Physiological Intracytoplasmic Sperm Injection (PICSI) technique in patient with high DNA fragmentation index (DFI) sperm samples. Summary answer Sperm selected by microfluidic sorting are associated with significant increase in day 3 grade A embryo development rate, clinical pregnancy rate over PICSI. What is known already DNA damage is unrecognisable in living sperm prior to insemination and an increased sperm DNA fragmentation index has been associated with lower fertilization rates, impaired embryo development and reduced pregnancy rates. Standard semen processing techniques are associated with centrifugation, which may induce reactive oxygen species and DNA damage. In strategies to minimize sperm DNA fragmentation, Physiological ICSI can relatively reduce sperm DNA fragmentation by 67.9% (Parmegiani et al., 2010) while new technique Microfluidic sperm sorter technique also demonstrate sperm selection with significantly reduced DNA damage. Study design, size, duration A prospective randomised study was conducted from 1st August 2019 to 31st December 2020. Two hundred patients were randomised by computer generated list and divided into 2 groups. Group A (n = 100) , in which sperm were processed by microfluidic sperm sorter (MFSS) while in group B (n = 100), sperm were selected by Physiological Intracytoplasmic Sperm Injection (PICSI) technique and morphologically normal motile sperm were injected by Intracytoplasmic sperm injection (ICSI) technique in all mature oocytes. Participants/materials, setting, methods The study period included all normozoospermic patients with high DNA fragmentation index (&amp;gt;25% ) while oligospermic, asthenozoospermic samples, patients with poor ovarian reserve and advanced age were excluded from the study. All A grade embryos were vitrified and transferred in frozen embryo replacement cycle. Both groups were compared on the basis of fertilisation rate, day 3 grade A embryo development rate , clinical pregnancy rate and miscarriage rate. Main results and the role of chance Cycle characteristics (female age, length of stimulation, gonadotrophin dose, number of oocytes and number of transferred embryos) were similar in both groups. Between the 2 groups, There was a significant increase observed in day 3 grade A embryo development rate (60% vs. 42%, p–0.016) and clinical pregnancy rate (62% vs. 46%, p–0.049), while no statistical significant difference observed in fertilisation rate (82% vs. 78%, p–0.80) and miscarriage rate ( 12% vs. 11%, p- 1). Limitations, reasons for caution: Larger randomised control studies are needed to strengthen these results. Wider implications of the findings: We have demonstrated that sperm sorted by microfluidic helps in selection of sperm with better DNA integrity over Physiological ICSI. Using it in routine practice can help in reducing the negative effect of reactive oxygen species and thus improve pregnancy rate and live birth rate. Trial registration number MCDH/2019/31

Association of Iron Status in Follicular Fluid with Pregnancy Outcomes in Infertile Women Undergoing IVF/ICSI

Iron status may influence the outcome of infertile women under the intracytoplasmic sperm injection (ICSI) technique of in vitro fertilization (IVF). The aim of this study is to evaluate iron status and ceruloplasmin ferroxidase activity in the follicular fluid (FF) and their association with IVF outcomes. The study enrolled fertile women with male cause infertility (n=25), infertile women with polycystic ovary syndrome (PCOS; n=21), infertile women with low AMH level (n=26), and women with unexplained infertility (UI; n=27), all undergoing IVF/ICSI. On the day of oocyte suction, the selection of FF samples was accomplished. Iron, ferritin, and transferrin levels, as well as ceruloplasmin (CP) ferroxidase activity, were measured in the FF. In the PCOS group, iron showed significantly higher level (P&lt;0.05) as compared to the control and UI groups. In the PCOS group, ferritin showed significantly higher level (P&lt;0.05) compared with the control group. In the PCOS group, transferrin showed significantly higher level (P&lt;0.05) when compared with the UI group. Also, Cp. ferroxidase activity in the PCOS group showed a lower level, but non-significant difference, compared with the other groups. In conclusion, the increased iron level in the follicular fluid of women with PCOS may lead to decrease pregnancy success after applying IVF protocol.

The Effect of Interleukin-10 And Its Relationship with The Level of White Blood Cells in Women Spontaneous Miscarriage Undergoing Intracytoplasmic Sperm Injection (ICSI) technique

This study was conducted on 84 samples of women subject to the ICSI program . Blood sample was taken tomeasure WBC count , Interleukin-10 and ?-hCG levels . Groups were divided based on the ?-hCG level totwo groups (Pregnant Women Group - Pregnancy Failure Group) and the total of pregnant women dividedinto (Pregnancy Group and spontaneous miscarriage) .The results of the current research showed a significant increase in the level ?-hCG and that the increasein the level of this hormone is evidence of the presence of high success rates for pregnancy in women whoperformed operations IVF, where the success rate at the beginning of the matter reached 61.9%, after whichit decreased to 33.3% after the first three months due to the occurrence of spontaneous miscarriage ofpregnant women due to various immunological and physiological reasons, as well as a positive correlationbetween the level of ?-hCG and other parameters within the study (Interleukin-10 -WBC).The results of the current research also showed a significant difference between the group (pregnancyfailure) and the group (spontaneous miscarriage) compared with the control group (continued pregnancy)in relation to the level of Interleukin-10, Also, The results of the current research showed a significantdifference between the group (pregnancy failure) and the group (spontaneous miscarriage) compared withthe control group (continuation of pregnancy) in relation to the level of WBC, and the present study found apositive relationship between the level of Interleukin-10 and WBC