Intracytoplasmic Sperm Injection Attempts Research Articles

First-line infertility treatment in normal or subnormal sperm: Interest of a simplified pre-IMSI test.

In the field of male infertility, when sperm is normal/subnormal, a few "add-on" routine tests can complete the basic semen examination. The aim of this study was to develop and evaluate a faster, simplified motile sperm organelle morphology examination (MSOME) technique for selected infertile patients with apparently normal/subnormal sperm and, in their background: failure of two or three intrauterine insemination (IUI) cycles, repeatedly fragmented embryos, embryonic development to blastocyst-stage failures, repeated miscarriages, a long period of infertility or 2 or more IVF attempts without pregnancy. Our test results were correlated with IUI, conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI) outcomes. We validated an adapted version of the MSOME analysis called the pre-IMSI test (PIT), based on vacuole evaluation alone. 248 infertile patients from our assisted reproductive technology (ART) Center were retrospectively selected and split into three PIT score subgroups (patients with ≤8% (score I), 9 to 15% (score II) and ≥16% normal spermatozoa (score III)) based on the correlation between PIT results and each ART technique outcome. The choice of one or another of these ART techniques had been made according to the usual clinico-biological criteria. Clinical outcomes for each of the three PIT subgroups were compared individually for the different ART techniques. For ICSI, the effect of the PIT score subgroup was significant for clinical pregnancies (p = 0.0054) and presented a trend for live births (p = 0.0614). Miscarriage rates of IVF attempts were statistically different depending on the PIT score (p = 0.0348). Furthermore, the odds ratios of clinical pregnancy rates were significantly different according to PIT score subgroup when comparing ICSI vs. IMSI or IVF vs. ICSI attempts. IMSI appears to be recommended when sperm belongs to PIT score I, ICSI when it belongs to PIT score II and IVF or IUI when sperm is of PIT score III quality in selected infertile couples. The lack of statistical power in these PIT subgroups means that we must remain cautious in interpreting results. Our results support the interest of this simplified test for certain couples with normal/subnormal sperm to help choose the most efficient ART technique, even as first-line treatment.

P-027 Sperm motility and calcium ionophore treatment influence fertilization and pregnancy outcomes in TESE-ICSI

Abstract Study question Are there differences in fertilization and pregnancy rates post-TESE-ICSI between motile and immotile sperm, and do these vary in treatments with and without calcium ionophore? Summary answer Post-TESE-ICSI data reveals differences in fertilization and pregnancy rates when comparing motile vs. immotile sperm and treatments with and without calcium ionophore after TESE-ICSI. What is known already Testicular sperm extraction (TESE) followed by intracytoplasmic sperm injection (ICSI) poses a challenge for every IVF laboratory. This is particularly true in patients exhibiting azoospermia, where only a few and mostly immotile sperm are present in testicular tissue. In these cases, a calcium ionophore treatment for activation of the oocyte may help to overcome poor prognosis fertilization outcome, and thereby enhancing the likelihood to achieve a transfer leading to a successful pregnancy. Although both techniques, TESE-ICSI and activating substances are commonly applied, detailed analyses for individual counselling of couples are missing. Study design, size, duration This study analyzed 787 TESE-ICSI cycles (2015-2023) regarding patient age, number of attempts, oocyte maturity, fertilization and pregnancy rate after fresh embryo transfer. Cycles were split into four groups: (A) at least one motile sperm was injected (n = 493); (B) only immotile sperm were used (n = 294); (C) calcium ionophore (n = 474); (D) no calcium ionophore (n = 313). 17 cycles were excluded from the study because no sperm were found on the day of TESE-ICSI and oocytes were vitrified. Participants/materials, setting, methods In our IVF clinic, calcium ionophore treatments are only offered if a pre-screening of TESE samples reveals the absence of motile spermatozoa or if a previously conducted TESE-ICSI attempt resulted in a low fertilization rate (<30%) or a total fertilization failure. Statistical analyses was performed to compare the motile and immotile sperm group and the groups with or without calcium ionophore treatment. Main results and the role of chance No differences were found in all groups (A-D) regarding age of female or male patient and number of collected oocytes per group. However, our results demonstrate a difference in the mean number of ICSI attempts when comparing motile sperm and immotile sperm (2.0±1.3 vs. 2.3±1.6, p < 0.05), and when comparing attempts with calcium ionophore treatment and those without (2.4±1.6 vs. 1.8±1.8, p < 0.0005). The mean fertilization rate (2PN stage) was higher in the motile sperm group (A) compared to the immotile group (B) (57.8% vs. 28.4%, p < 0.0005) and also higher in attempts of group (D) in comparison to attempts of group C (56.8% vs. 40.2%, p < 0.0005). While fertilization failure occurred less often in group (A) (3.2%) it occurred to a higher rate in group (B) (19.4%), followed by group (C) (12.0%). Biochemical (BPR) and clinical pregnancy rates (CPR) showed a significant difference between the motile and immotile sperm group (BCR: 33.2% vs. 12.3%, p < 0.0005; CPR: 25.6% vs. 9.3%, p < 0.0005), but no differences between the two calcium ionophore groups. Limitations, reasons for caution This analysis was done retrospectively. A potential subgroup analysis of groups (A) and (B) into calcium ionophore use and of groups (C) and (D) into motile or immotile sperm will reveal the effect of calcium ionophore on success rates regardless of sperm motility. Wider implications of the findings Our data provides insight into success rates of TESE-ICSI cycles, especially if only immotile sperm were present. This will serve as a helpful guide for physicians when counseling patients about their success rates and to optimize the outcome in cases with poor prognosis by using a calcium ionophore treatment. Trial registration number not applicable

Results and perinatal outcomes from 189 ICSI cycles of couples with asthenozoospermic men and flagellar defects assessed by transmission electron microscopy

Research questionDo patients presenting with flagella ultrastructural defects as assessed by electron microscopy, and defined within three phenotypes (dysplasia of the fibrous sheath [DFS], primary flagellar dyskinesia [PFD] and non-specific flagellar abnormalities [NSFA]), have decreased chances of success in intracytoplasmic sperm injection (ICSI) or adverse obstetric and neonatal outcomes? DesignRetrospective analysis of 189 ICSI cycles from 80 men with spermatozoa flagellum ultrastructural defects (DFS [n = 16]; PFD [n = 14]; NSFA [n = 50] compared with a control group (n = 97). Cycles were cumulatively analysed. All fresh and frozen embryo transfers resulting from each ICSI attempt were included. The effect of transmission electron microscopy (TEM) phenotype on the main ICSI outcomes was assessed by a multivariate logistic regression combined with a generalized linear mixed model to account for the non-independence of the observations. ResultsNo predictive value of TEM phenotype was found on the main outcomes of ICSI, namely fertilization rates, pregnancy and delivery rates, and cumulative pregnancy and delivery rates. Cumulative pregnancy rates ranged from 29.0–43.3% in the different TEM phenotype subgroups compared with 36.8% in the control group. Cumulative live birth rates ranged from 24.6–36.7% compared with 31.4% in the control group. No increase was found in miscarriages, preterm births, low birth weights or birth abnormalities. ConclusionsData on the cumulative chances of success in ICSI of patients with ultrastructural flagellar defects, a rare cause of male infertility often associated with an underlying genetic cause, are reassuring, as are obstetrical and neonatal outcomes in this population.

Successful cryptozoospermia management with multiple semen specimen collection

To determine the prevalence of sperm suitable for intracytoplasmic sperm injection (ICSI) in fresh ejaculated semen samples provided by men scheduled for a microdissection testicular sperm extraction (mTESE) procedure. Secondary objectives included an evaluation of the effect of a short abstinence period on semen quality and ICSI outcomes for men with cryptozoospermia. Retrospective cohort study. Academic medical center. All men were scheduled to undergo a mTESE procedure by a single, high-volume surgeon at an academic center from September 1, 2015, to May 1,2021. Presence of sperm suitable for ICSI in the ejaculate on the day of scheduled mTESE. Prevalence of sperm suitable for ICSI in the ejaculate among previously diagnosed men with azoospermia. Secondary outcomes included changes in semen parameters, clinical pregnancy rate, and live birth rate. Of 727 planned mTESE procedures, 69 (9.5%) were canceled because sperm suitable for ICSI were identified in a fresh ejaculated sample produced on the day of scheduled surgery (typically one day before oocyte retrieval). Overall, 50 men (50/727, 6.9%) used these rare, ejaculated sperm for ICSI. Semen samples obtained with <24 hours of abstinence were more likely to have better motility than the sample initially provided on the day of the planned mTESE. The live birth rate per ICSI attempt using these rare, ejaculated sperm was 36% (19/53). Providing a fresh ejaculated semen sample on the day of mTESE allows nearly 10% of men with azoospermia to avoid surgery with satisfactory ICSI outcomes. Providing multiple ejaculated samples over a short period of time does not adversely affect sperm concentration and may enhance sperm motility in men with cryptozoospermia.

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

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

Hyaluronic acid-sperm selection significantly improves the clinical outcome of couples with previous ICSI cycles failure.

Beneficial effects of hyaluronic acid (HA)-based selection of spermatozoa for intracytoplasmic sperm injection (ICSI) are still controversial, and further studies are needed to categorize patients that might benefit from such a method. We investigated whether HA sperm selection improved ICSI outcome of couples with previous ICSI cycle failure. In this retrospective study, we prospectively collected data of (i) Group 1: 96 couples who performed one failed ICSI cycle ("1st procedure," n=96) followed by another ICSI cycle ("2nd procedure," n=101); ii) Group 2: 87 couples who performed one failed ICSI cycle (n=87) followed by an HA-ICSI cycle (n=104). Differences between procedures and groups were measured by paired and independent statistical tests, respectively. A generalized linear mixed model analyzed the effect of procedure on the outcomes and the interaction between procedures and groups. Injection of HA-bound sperm significantly improved cleavage rate with respect to standard ICSI (p=0.026). No evolutive pregnancies were obtained in the 1st ICSI attempts. The 2nd ICSI cycles resulted in successfully seven pregnancies. In HA-ICSI cycles, the better quality of embryos with respect to ICSI (p=0.034) increased the choice of day 5 embryo transfer (p=0.030), which resulted in successfully 28 pregnancies. No differences were observed in clinical outcomes of the two ICSI procedures in Group 1, while pregnancy and implantation rates were significantly higher in HA-ICSI with respect to ICSI cycles (p=0.001, p<0.0001, respectively). No negative perinatal outcomes were recorded. In couples where previous 1st ICSI failed, selection of HA-bound spermatozoa significantly improved clinical outcomes with respect to further standard ICSI. This study identified couples with previous ICSI cycles failure as a category of infertile patients that really may benefit from HA sperm selection before ICSI.

P–143 Improved embryonic development and utilization rates with EmbryoScope: A within-subject comparison versus a benchtop incubator

Abstract Study question In consecutive intracytoplasmic sperm injection (ICSI) cycles, is embryonic development in EmbryoScope better than the previous one obtained in a benchtop (G–185) incubator? Summary answer Embryonic development, and oocyte and embryo utilization rates (OUR and EUR) are significantly improved in the EmbryoScope, as compared to G–185. What is known already The time-lapse imaging (TLI) system, which allows a non-invasive continuous assessment of embryo morphokinetics parameters in a closed culture system has been developed, promising improved embryo development by reducing oscillations in pH, humidity and temperature. To investigate this hypothesis, one study has already compared embryonic development in a TLI versus a benchtop incubator. However, It has never been investigated whether embryonic development can be improved within-subject, by changing from benchtop incubator in the first intracytoplasmic sperm injection (ICSI) cycle to the EmbryoScope, a TLI incubator, in the following ICSI cycle, and that was the objective of the present study. Study design, size, duration This study had a retrospective within-subject design, in which each cycle served as its own control. Data were obtained via chart review of patients undergoing ICSI in a private university–affiliated IVF center that fulfilled the following criteria: second ICSI attempt in which embryos had been cultured in a TLI incubator system (TLI group, n = 71), preceded by a first ICSI attempt in which embryos had been cultured in a conventional incubator (Control group, n = 71). Participants/materials, setting, methods Embryonic development up to the fifth day of development, OUR (transferred embryos plus frozen embryos per retrieved oocytes) and EUR (transferred embryos plus frozen embryos per fertilized oocytes) were compared between the groups using generalized linear models followed by Bonferroni post hoc. The post hoc achieved power was 82.6%, considering the sample size, the effect size obtained for blastocyst development rate and 5% significance level. Main results and the role of chance There were significant differences in fertilization rate (76.0% ± 1.3 vs. 80.0% ± 1.4, p = 0.044, OR: 1.051, CI: 1.001 – 1.103), non-fertilization rate (14.8% ± 0.6 vs. 6.3% ± 0.4, p &amp;lt; 0.001, OR: 0.424, CI: 0.370 – 0.486), day–2 non-cleavage rate (3.8% ± 0.2 vs. 1.1% ± 0.1, p &amp;lt; 0.001, OR: 0.285, CI: 0.234 – 0.347), blastocyst development rate (40.9% ± 1.1 vs. 55.6% ± 1.3, p &amp;lt; 0.001, OR: 1.358, CI: 1.267 – 1.456), frozen blastocyst rate (31.8% ± 0.8 vs. 37.0% ± 0.9, p &amp;lt; 0.001, OR: 1.163, CI: 1. 085 – 1.248), OUR (40.7% ± 1.0 vs. 50.2% ± 1.1, p &amp;lt; 0.001, OR: 1.232, CI: 1.155 – 1.314), and EUR (52.4% ± 1.1 vs. 66.6% ± 1.2, p &amp;lt; 0.001, OR: 1.269, CI: 1.202 - 1.341), all in favor of TLI group. Pregnancy rate (30.2% vs. 30.8%, p = 0.940), implantation rate (24.6% ± 40.0 vs. 26.1% ± 41.6, p = 0.830), and miscarriage rate (21.1% vs. 15.0%, p = 0.622) were similar between Control and TLI groups, respectively. Limitations, reasons for caution (i) Different culture dishes were used in each system; (ii) it is not possible to confirm how much of the embryonic improvement was due to the culture conditions; (iii) the study design is not ideal for the comparison of clinical outcomes and, also, underpowered to do so. Wider implications of the findings: Even though the clinical outcomes were similar between the groups, the results may also lead to higher cumulative pregnancy outcomes following embryo thawing and transfer. Trial registration number Not applicable

Oral Antioxidant Treatment of Men Significantly Improves the Reproductive Outcome of IVF Cycles.

Some 30% to 80% of male sub-fertility may be associated with oxidative stress that damages spermatozoa and can decrease success of in vitro fertilization (IVF) techniques. This multicenter, longitudinal, prospective study aimed to investigate whether oral antioxidant supplementation improved the reproductive competence of men who had had low fertilization rates in their previous intracytoplasmic sperm injection (ICSI) cycles without azoospermia or severe oligozoospermia or any identifiable andrological disease. Seventy-seven men from couples who had an ICSI attempt with unexplained <60% fertilization rate took an antioxidant mix of myo-inositol, alpha-lipoic acid, folic acid, coenzyme Q10, zinc, selenium, and vitamins B2, B6, and B12. Semen parameters were analyzed before (T0) and after 90 days (T90) of treatment, and outcomes of the paired T0 and T90 cycles were compared. After the treatment there was an increase in sperm concentration (p = 0.027), total motile sperm count (p = 0.003), progressive motility (p < 0.0001), and a decreasing trend of DNA-fragmented spermatozoa. Embryological outcomes (fertilization, embryo quality, blastocyst development) were significantly higher in T90 than T0 cycles. No T0 cycle resulted in an evolutive pregnancy. Conversely, in T90 cycles 29 singleton clinical pregnancies were obtained. No negative neonatal outcomes were recorded in newborns after antioxidant treatment. Diet supplementation of men who have had low fertilization rates in their previous ICSI cycles with a combination of myo-inositol, alpha-lipoic acid, folic acid, coenzyme Q10, zinc, selenium, betaine, and vitamins may improve semen reproductive potential and ICSI clinical outcome.

Identification of Novel Biallelic TLE6 Variants in Female Infertility With Preimplantation Embryonic Lethality.

Preimplantation embryonic lethality is a rare cause of primary female infertility. It has been reported that variants in the transducin-like enhancer of split 6 (TLE6) gene can lead to preimplantation embryonic lethality. However, the incidence of TLE6 variants in patients with preimplantation embryonic lethality is not fully understood. In this study, we identified four patients carrying novel biallelic TLE6 variants in a cohort of 28 patients with preimplantation embryonic lethality by whole-exome sequencing and bioinformatics analysis, accounting for 14.29% (4/28) of the cohort. Immunofluorescence showed that the TLE6 levels in oocytes from patients were much lower than in normal control oocytes, suggesting that the variants result in the lower expression of the TLE6 protein in oocytes. In addition, a retrospective analysis showed that the four patients underwent a total of nine failures of in vitro fertilization and intracytoplasmic sperm injection attempts, and one of them became pregnant on the first attempt using donated oocytes. Our study extends the genetic spectrum of female infertility caused by variants in TLE6 and further confirms previously reported findings that TLE6 plays an essential role in early embryonic development. In such case, oocyte donation may be the preferred treatment.

Intracytoplasmic sperm injection versus conventional in-vitro fertilisation in couples with infertility in whom the male partner has normal total sperm count and motility: an open-label, randomised controlled trial

The use of intracytoplasmic sperm injection has increased substantially worldwide, primarily in couples with non-male factor infertility. However, there is a paucity of evidence from randomised trials supporting this approach compared with conventional in-vitro fertilisation (IVF). We aimed to investigate whether intracytoplasmic sperm injection would result in a higher livebirth rate compared with conventional IVF. This open-label, multicentre, randomised trial was done at two IVF centres in Ho Chi Minh City, Vietnam (IVFMD, My Duc Hospital and IVFAS, An Sinh Hospital). Eligible couples were aged at least 18 years and the male partner's sperm count and motility (progressive motility) were normal based on WHO 2010 criteria. Couples had to have undergone two or fewer previous conventional IVF or intracytoplasmic sperm injection attempts, have used an antagonist protocol for ovarian stimulation, and agree to have two or fewer embryos transferred. Couples were randomly assigned (1:1) to undergo either intracytoplasmic sperm injection or conventional IVF, using block randomisation with variable block size of 2, 4, or 8 and a telephone-based central randomisation method. The computer-generated randomisation list was prepared by an independent statistician who had no other involvement in the study. Embryologists and couples were not masked to study groups because of the type of interventions and differences in hospital fees, but clinicians performing embryo transfer were unaware of study group allocation. The primary outcome was livebirth after the first embryo transfer from the initiated cycle. Analyses were done on an intention-to-treat basis. The trial is registered with ClinicalTrials.gov, NCT03428919. Between March 16, 2018, and Aug 12, 2019, we randomly assigned 1064 couples to intracytoplasmic sperm injection (n=532) or conventional IVF (n=532). Livebirth after the first embryo transfer from the initiated cycle occurred in 184 (35%) of 532 couples randomly assigned to intracytoplasmic sperm injection and in 166 (31%) of 532 couples randomly assigned to conventional IVF (absolute difference 3·4%, 95% CI -2·4 to 9·2; risk ratio [RR] 1·11, 95% CI 0·93 to 1·32; p=0·27). 29 (5%) couples in the intracytoplasmic sperm injection group and 34 (6%) couples in the conventional IVF group had fertilisation failure (absolute difference -0·9%, -4·0 to 2·1, RR 0·85, 95% CI 0·53 to 1·38; p=0·60). In couples with infertility in whom the male partner has a normal total sperm count and motility, intracytoplasmic sperm injection did not improve the livebirth rate compared with conventional IVF. Our results challenge the value of the routine use of intracytoplasmic sperm injection in assisted reproduction techniques for this population. My Duc Hospital and Merck Sharp and Dohme.

Circulating MMP-7 and VEGF as potential predictive biomarkers for recurrent implantation failures.

Recurrent implantation failure (RIF) is considered to be one of the major limiting factors of assisted reproductive technology (ART) programme success. The current study focused on the investigation of matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), cytokines and cell adhesion molecules in peripheral blood (PB) and follicular fluid (FF) obtained from 44 women aged between 25 and 39 years old and undergoing intracytoplasmic sperm injection (ICSI). These women were divided into two groups: 22 RIF women with embryo implantation failures after the transfer of at least four fresh or frozen-thawed good quality embryos in a minimum of three ICSI cycles, and 22 ICSI success women (controls) who achieved a clinical pregnancy at their first ICSI attempt. The PB and FF samples were obtained from each patient on the day of oocyte retrieval. MMP-1, -2, -3, -7, -9, TIMP-1, -2, vascular endothelial growth factor (VEGF), leukaemia inhibitory factor (LIF), vascular cell adhesion molecule 1 (VCAM1) and intercellular adhesion molecules 1 (ICAM1) were analyzed using enzyme-linked immunosorbent assay of PB and FF. Our results showed significant decreases in PB MMP-7 and PB VEGF in the RIF group compared with controls [281.11 (33-614) pg/ml vs 119.92 (27-441) pg/ml; P-value = 0.030] and [82.54 (25.94-210.20) pg/ml vs 30.93 (13.62-193.33) pg/ml; P-value = 0.022; respectively]. Receiver operating characteristic (ROC) curve analysis showed informative area under the curve values for PB MMP-7, as well as for PB VEGF, making them able to be proposed as biomarkers of the RIF. Therefore, circulating MMP-7 and VEGF seem to play an interesting role in embryo implantation in in vitro fertilization (IVF)/ICSI cycles and could be proposed as circulating biomarkers of the RIF. These results could be helpful for clinicians and patients to choose the best rescue strategy and treatment to minimize implantation failure in women undergoing IVF/ICSI procedures after the first attempt.

Does bacterial colonization during embryo transfer have an impact on pregnancy rate in ICSI? : Tunisian preliminary results

IntroductionEmbryos’ transfer into the uterine cavity remains a limiting factor in the success of intracytoplasmic sperm injection (ICSI). This study aimed to evaluate the impact of cervico-vaginal colonization on ICSI outcomes. Materiel and methodsLongitudinal, prospective and analytical study, conducted from February 1 st to October 31 st, 2017. On the day of embryos’ transfer, patients with the following criteria were included: Age ≤ 40, normal uterine ultrasound and hysteroscopy, first or second attempt of ICSI, use of fresh semen and transfer of at least one good quality embryo. Three samples were done for microbiological analysis: vaginal secretions, cervical mucus and the tip of the transfer catheter. ResultsThe study included 40 patients. Pregnancy rate was 52.5 %. Catheter contamination occurred in nine cases (22.5 %). The most frequently isolated bacteria were Streptococcus anginosus (37 % of cases), Gardnerella vaginalis (27 % of cases) and Streptococcus agalactiae (18 % of cases). In all cases of contamination, the same bacteria were found also in the vagina and cervical mucus. In cases of contamination with Gardnerella vaginalis, the study of vaginal flora showed bacterial vaginosis. No pregnancy was obtained when contamination of the transfer catheter occurred. In the absence of contamination, pregnancy rate was 61.2 % (ρ = 0.003). Both populations (patients with catheter contamination and patients without catheter contamination) were comparable regarding epidemiological characteristics and ICSI attempt data. ConclusionContamination of the transfer catheter by cervical-vaginal bacteria appears to affect IVF results. Systematic antibiotic prophylaxis could be proposed to reduce this negative impact.

Intracytoplasmic sperm injection outcomes after anti-oxidant treatment in repeated implantation failure

Objectives: Aim of this study was to explore the effects of three months oral anti-oxidant treatment in normozoospermic men with high DNA fragmentation index (DFI) and recurrent intracytoplasmic sperm injection (ICSI) failures. Methods: This is a retrospective analyses of normozoospermic patients with high DFI levels (&amp;gt; 30%) with at least two unsuccessful ICSI attempts. Among the participants, the study group included men who had oral anti-oxidant treatment. The men who did not receive any treatment during the study period were taken as controls. The outcomes of new ICSI cycles were compared. Results: During the study period 24 men have used three months of oral anti-oxidant treatment before a new ICSI cycle whereas the remaining 46 had ICSI-only without any other intervention. Duration of stimulation, the number of metaphase II oocytes, fertilization rate and implantation rates were similar between two groups. Although not statistically significant, cases in anti-oxidant group have slightly better pregnancy rates compared to ICSI-only group (37.5% vs 19.6%, p &amp;gt; 0.05). Only one case in each group had abortion. Conclusions: This study reveals that there is no statistically significant difference in the ICSI outcomes of the patients with high DFI levels, following unsuccessful ICSI attempts after oral anti-oxidant treatment. The possible benefit derived from the anti-oxidant therapy needs to be clarified with clinical studies with greater study populations.

Assessment of CD56+ CD69+ Natural Killer Cells in the Peripheral Blood of Women with Repeated Implantation Failure (RIF) undergoing an Intra Cytoplasmic Sperm Injection (ICSI) Cycle

Background: infertility is an increasing medical and social problem. Delayed childbearing exposes couples to increased risk of causes of infertility including endometriosis, pelvic infection, and poor egg and sperm quality. Many infertile couples turn to in-vitro fertilization (IVF). Objective: to analyze and compare between the peripheral blood NK cells (CD56+ CD69+ ) in women with repeated ICSI failure. Patients and Methods: the current study is a case control study. One hundred women were included and divided into two groups: The study group included 50 women with history of unexplained infertility and repeated implantation failure in their ICSI attempts and a control group which included 50 normal, non-pregnant and fertile women. Results: the current study revealed that there was a highly statistically significant difference between control and case groups regarding CD3- CD56+dim NK percentage with a p-value <0.0001. The current study revealed that there was a statistically significant difference between control and case groups regarding CD3- CD56+dim CD69+ NK cells percentages with a p-value 0.001. Conclusion: there was a highly statistically significant difference between control and case groups regarding CD3- CD56+dim NK percentage. Also the study showed that total CD56+dim NK had a very good predictive value as it had a sensitivity of 80.0%, a specificity of 66.7%, a PPV of 73.3% and a NPV of 78.8%.

Assisted oocyte activation significantly increases fertilization and pregnancy outcome in patients with low and total failed fertilization after intracytoplasmic sperm injection: a 17-year retrospective study

To investigate the extent to which assisted oocyte activation (AOA) improves clinical outcomes in patients diagnosed with oocyte activation deficiencies (OADs). Retrospective cohort study comparing AOA cycles and previous intracytoplasmic sperm injection (ICSI) cycles in couples experiencing low or total failed fertilization after ICSI. Importantly, the sperm-related oocyte-activating capacity was examined in all patients before AOA with the use of the mouse oocyte activation test (MOAT). Infertility center at a university hospital. A total of 122 couples with a history of low or total failed fertilization after ICSI. ICSI, MOAT, AOA, and embryo transfer. Fertilization, pregnancy, and live birth rates. MOAT revealed 19 patients with a sperm-related OAD (MOAT group 1), 56 patients with a diminished sperm-related oocyte-activating capacity (MOAT group 2), and 47 patients with a suspected oocyte-related OAD (MOAT group 3). AOA (191 cycles) significantly improved fertilization, pregnancy, and live birth rates in all MOAT groups compared with previous ICSI attempts (243 cycles). Fertilization rates after AOA were significantly different among MOAT groups 1 (70.1%), 2 (63.0%), and 3 (57.3%). Between MOAT group 1 and 3, significant differences in pregnancy (49.0% vs. 29.4%) and live birth (41.2% vs. 22.1%) rates were observed. In total, 225 embryo transfers resulted in 60 healthy live births following AOA. Patients undergoing diagnostic testing before AOA show a significant improvement in clinical outcomes compared with previous cycles. Our findings highlight that AOA should be reserved for patients with clear OADs.

Association between progesterone to number of mature oocytes index and live birth in GnRH antagonist protocols.

What is the association between blood progesterone level alone, progesterone to mature oocytes index (PMOI) and IVF outcomes? Clinical data from 960 couples undergoing their first fresh embryo transfer after an intracytoplasmic sperm injection attempt carried out between September 2012 and July 2017 were analysed. All patients underwent ovarian stimulation combining recombinant FSH and gonadotrophin releasing hormone antagonist. Progesterone was measured on the day on which ovulation was triggered. The PMOI was divided into four groups based on 25th, 50th and 75th percentiles, and live birth and implantation rates were compared between the groups. A negative association was found between PMOI levels, live birth and implantation rates. When adjusting for age, ovarian stimulation index (OSI) and number of embryos transferred, the PMOI remained negatively correlated to live birth rate (LBR) (OR = 0.147 [0.031 to 0.701]; P = 0.0161), whereas total blood progesterone was no longer evident. Moreover, the LBR (18.5% versus 28.4%; P < 0.01) and implantation rate (12.4% versus 21.0%; P < 0.01) were significantly decreased only when PMOI was 0.167 ng/ml or greater, irrespective of progesterone concentration (< or ≥1.08 ng/ml). The opposite was not true, however. Similar PMOIs were recorded in the same patient from one attempt to the next, and were partially linked to basal FSH, anti-Müllerian hormone, antral follicle count and OSI. PMOI seems to be more predictive than total progesterone level of IVF outcome and reflects the ability of embryos to develop as it is linked to the implantation rate.

DNA fragmentation index and human papilloma virus in males with previous assisted reproductive technology failures.

This study was designed to evaluate the prevalence of Human Papilloma Virus (HPV) in semen and document the cycle outcomes in couples with previous intra-cytoplasmic sperm injection (ICSI) failures. One hundred and seventeen couples with at least two ICSI attempts were included in the study. HPV infection in semen and DNA fragmentation in samples were analyzed by commercially available kits. The percentage of spermatozoa with fragmented DNA (DNA fragmentation index: DFI) was determined during fluorescence microscopic examination as previously described. The cycle outcomes of couples with or without HPV infected male partners were recorded. According to our results, the prevalence of HPV was 7.7% in asymptomatic males with at least two previous ICSI failures. The increased DFI (>30%) was observed in 82.9% of the cases. In HPV-positive cases significantly lower number of good quality embryos were obtained. The implantation and pregnancy rates were similar in infected and non-infected males (p>0.05). The early miscarriage rate was slightly higher in HPV- positive group (33% vs. 10%, p>0.05). In cases with previous ICSI failures, the prevalence of HPV infection in semen is not higher than previously reported infertile populations. The reproductive outcome might be impaired in HPV-positive semen due to lower number of good quality embryos, which needs to be clarified by further large population-based studies.

Homozygous Mutations in WEE2 Cause Fertilization Failure and Female Infertility

Fertilization is a fundamental process of development and is a prerequisite for successful human reproduction. In mice, although several receptor proteins have been shown to play important roles in the process of fertilization, only three genes have been shown to cause fertilization failure and infertility when deleted invivo. In clinical practice, some infertility case subjects suffer from recurrent failure of invitro fertilization and intracytoplasmic sperm injection attempts due to fertilization failure, but the genetic basis of fertilization failure in humans remains largely unknown. Wee2 is a key oocyte-specific kinase involved in the control of meiotic arrest in mice, but WEE2 has not been associated with any diseases in humans. In this study, we identified homozygous mutations in WEE2 that are responsible for fertilization failure in humans. All four independent affected individuals had homozygous loss-of-function missense mutations or homozygous frameshift protein-truncating mutations, and the phenotype of fertilization failure was shown to follow a Mendelian recessive inheritance pattern. All four mutations significantly decreased the amount of WEE2 protein invitro and in affected individuals' oocytes invivo, and they all led to abnormal serine phosphorylation of WEE2 and reduced tyrosine 15 phosphorylation of Cdc2 invitro. In addition, injection of WEE2 cRNA into affected individuals' oocytes rescued the fertilization failure phenotype and led to the formation of blastocysts invitro. This work presents a novel gene responsible for human fertilization failure and has implications for future therapeutic treatments for infertility cases.

Can intracytoplasmic Morphologically Selected Spermatozoa Injection be Used as First Choice of Treatment for Severe Male Factor Infertility Patients?

Aim:This study was carried out to assess the outcome of the intracytoplasmic morphologically selected sperm injection (IMSI) technique compared with the previous failed intracytoplasmic sperm injection (ICSI) attempts in oligoasthenoteratozoospermia (OAT)/severe OAT (SOAT)/teratozoospermia patients.Setting:Institution-based, in vitro fertilization center.Study Design:It was a nonrandomized prospective study including 57 couples who had previous one or two ICSI failures (failure due to no implantation as embryos were transferred in these cycles and had no pregnancy) due to male factor. There was no case of total fertilization failure. IMSI was carried out in these couples and the results were compared with their previously failed ICSI attempts.Materials and Methods:Real-time selection of sperms was done using IMSI as it allows the assessment of fine nuclear morphology and vacuoles in the sperm head at a high magnification (>6000×) with differential interference contrast optics. Therefore, IMSI was applied in couples having OAT, SOAT or teratozoospermia as male factor and the results were compared with their previous failed ICSI attempts. Statistical analysis was carried out using GraphPad Prism.Results and Conclusion:Data analysis demonstrated significant difference in the fertilization rate between IMSI and previous ICSI attempts of these patients (30.0% vs. 52.0%; P < 0.05). The embryo quality, implantation and pregnancy rates with IMSI were also significantly higher than those of their previous ICSI cycles (32% vs. 56.4%; 30.2% vs. 68.5%; 0.0% vs. 62.4%; P < 0.05). Our conclusion is that the IMSI procedure improved embryo development and the clinical outcomes in the same infertile couples with male infertility and poor embryo development over their previous ICSI attempts and can be taken up as the treatment of choice in cases of severe male factor infertility.

Microdissection testicular sperm extraction in Finland - results of the first 100 patients.

Testicular microdissection sperm extraction (MD-TESE) combined with intracytoplasmic sperm injection (ICSI) has made biological fatherhood possible for many men with the most severe form of male infertility, non-obstructive azoospermia. MD-TESE was introduced in Turku in 2008, and by 2015, 100 Finnish men with non-obstructive azoospermia have been operated on. The average age of the men was 33years at the time of surgery. Forty-eight had a needle biopsy previously and 56% had a testicular size <15mL. The most common diagnoses were idiopathic (n=65), Klinefelter syndrome (n=15), operated cryptorchidism or torsion (n=10), and Y chromosome microdeletion (n=7). The pregnancy outcomes were followed. The sperm recovery rate (SRR) overall was 42%: 31% for idiopathic non-obstructive azoospermia, 40% for Klinefelter syndrome, 57% for Y chromosome microdeletion AZFc, 90% for previous testicular surgery (mostly for cryptorchidism; n=10) and 67% for previous cytotoxic treatment (n=3). SRR with histopathologic diagnosis Sertoli-cell-only was 29%, and 44% for spermatogenic arrest. Age did not affect the outcome of the surgery. Small testicular size seemed to predict a higher SRR. A previous needle biopsy did not predict a lower SRR. Surgical complications were rare. Of couples, 32 had at least one ICSI attempt, and 22 at least one live birth, giving a cumulative live birth rate of 69%. No major pregnancy complications occurred. Our SRR is comparable with international results, and the cumulative live birth rate similar to other ICSI indications in Finland. Physicians and specialists need to be made aware of new treatment options to enable biological fatherhood for men with non-obstructive azoospermia.