Improve In Vitro Fertilization Outcomes Research Articles

Melatonin supplementation attenuates cuproptosis and ferroptosis in aging cumulus and granulosa cells: potential for improving IVF outcomes in advanced maternal age

BackgroundAdvanced maternal age is associated with decreased oocyte quantity and quality and in vitro fertilization (IVF) success rates. This study aimed to investigate whether melatonin supplementation can improve IVF outcomes in women of advanced maternal age by modulating cuproptosis and ferroptosis.MethodsThis prospective cohort study included 161 women aged 35–45 years undergoing IVF-frozen embryo transfer cycles. Participants were assigned to either melatonin (n = 86, 2 mg daily for ≥ 8 weeks) or control (n = 75) groups. Cumulus cells were analyzed for cuproptosis and ferroptosis-related gene expression. Additional experiments were conducted on the HGL5 human granulosa cell line to assess mitochondrial function and metabolic reprogramming.ResultsMelatonin supplementation significantly improved IVF outcomes in women aged ≥ 38 years, increasing clinical pregnancy rates (46.0% vs. 20.3%, P < 0.01), ongoing pregnancy rates (36.5% vs. 15.3%, P < 0.01), and live birth rates (33.3% vs. 15.3%, P < 0.05). In cumulus cells from patients, gene expression analysis revealed that melatonin modulated cuproptosis and ferroptosis-related genes, including ATP7B and GPX4, with more pronounced effects in the ≥ 38 years group. This suggests melatonin enhances cellular resilience against oxidative stress and metal-induced toxicity in the ovarian microenvironment. In vitro studies using HGL5 cells showed melatonin reduced oxidative stress markers, improved mitochondrial function, restored expression of glycolysis and TCA cycle-related genes and modulated cuproptosis and ferroptosis-related gene expression. These findings provide mechanistic insight into melatonin’s protective effects against regulated cell death in ovarian cells, potentially explaining the improved IVF outcomes observed.ConclusionsMelatonin supplementation significantly improved IVF outcomes in women of advanced maternal age, particularly those ≥ 38 years old, likely by modulating cuproptosis and ferroptosis and enhancing mitochondrial function in cumulus and granulosa cells. These results suggest that melatonin could be a promising adjuvant therapy for improving IVF success rates in older women.

Detoxification as a potential intervention for improving in vitro fertilization outcomes: a comparative analysis of failed cycles before and after detox

Objectives: In vitro fertilization (IVF) success rates vary, posing challenges for patients with repeated failed cycles. Environmental toxins, including endocrine-disrupting chemicals, may contribute to IVF failure. Detoxification has emerged as a potential intervention to improve outcomes, but its efficacy remains uncertain. Methods: We conducted a retrospective analysis of data from 65 IVF patients undergoing detoxification at a private fertility clinic in Lagos, Nigeria, between 2020 and 2022. Demographic data, IVF history, and detoxification details were collected. Logistic regression assessed the impact of age and failed IVF cycles on live birth outcomes, with odds ratios calculated. Results: Of 65 patients, 33 achieved a positive pregnancy outcome after detoxification. A significant reduction in failed IVF cycles postdetoxification was observed (P &lt; 0.0001). Logistic regression indicated that older age and more failed IVF cycles were associated with decreased odds of live birth. However, detoxification mitigated this effect, reducing the odds of failed IVF cycles, although not significantly. Conclusion: Detoxification shows promise as an intervention to improve IVF outcomes in patients with repeated failed cycles. By reducing failed IVF cycles and enhancing live birth rates, detoxification may optimize reproductive success. Further research with larger sample sizes is needed to confirm these findings and elucidate underlying mechanisms. Nonetheless, our study highlights the importance of considering environmental factors and adopting holistic approaches to enhance IVF success.

The efficacy of orally administered L-carnitine in alleviating ovarian dysfunctions has laid the foundation for targeted in vivo use: a study employing self-control and propensity score matching.

This study aimed to evaluate the effectiveness of oral L-carnitine administration in patients after treatment failure to lay the groundwork for targeted in vivo use. A total of 515 In Vitro Fertilization (IVF) patients undergoing subsequent cycles were included after applying exclusion criteria. They were divided into a control group of 362 patients and a study group of 153 patients who received oral L-carnitine until oocyte retrieval.140 patients were matched according to maternal age, infertility duration, body mass index (BMI), day three top-quality embryos rate, by propensity score matching (PSM). The study investigated the relationship between L-carnitine treatment and in vivo oocyte maturation, normal fertilization, and subsequent embryo development. Following PSM, initial differences in BMI and Day3 top-quality embryo rate between groups were nullified, we created two comparable cohorts with highly similar characteristics. In the subsequent cycles, the study group showed significant improvements in in vivo oocyte maturation rate at retrieval (p=0.002), normal in vitro fertilization rate (p=0.003), blastocyst formation rate (p=0.003), and usable blastocyst rate compared to controls. Although there was no significant difference in the top-quality embryo rate on Day 3, the study group showed a 10% increase in the upper quartile (55.35% vs. 66.67%). The cumulative clinical pregnancy and live birth rates showed a significant improvement (59.82% vs. 68.42%,p=0.004, 47.41% vs. 59.80%, p=0.002). Furthermore, self-control analysis revealed substantial enhancements (p<0.001) in all outcome measures following L-carnitine administration, resulting in the birth of 74 healthy neonates without congenital anomalies. We theorized that daily oral intake of L-carnitine before oocyte retrieval could boost oocyte quality and embryonic development, thus improving IVF outcomes. Ongoing investigations hold the potential to offer valuable insights into the applications and mechanisms underlying the therapeutic effectiveness of L-carnitine.

Impact of Hysteroscopic Polypectomy on IVF Outcomes in Women with Unexplained Infertility.

Objective: To assess the effect of hysteroscopic polypectomy on the in vitro fertilization (IVF) results in infertile women with at least one prior negative IVF outcome. Methods: This retrospective cohort study included women who had attended the "2nd Department of Obstetrics and Gynecology of the National and Kapodistrian University of Athens" and "Iaso" Maternity Hospital from October 2019 to January 2023 for infertility treatment. The medical records of 345 women aged 18-45 years old without abnormal findings in hysterosalpingography (HSG) and with at least one previous failed IVF procedure were analyzed. The male factor was excluded, as well as a prior hysteroscopic removal of polyps. In 67 women, polyps were suspected during initial two-dimensional ultrasound (2D-US) examination. The final sample of the study comprised 40 patients, in which endometrial polyps were removed by hysteroscopy with the use of resectoscope. All patients underwent ovarian stimulation and IVF in the consecutive cycle using a short GnRh antagonist protocol. Main Results: After hysteroscopic polypectomy, 29 (72.5%) out of 40 patients had a positive pregnancy result: 26 (65%) clinical and 3 (7.5%) biochemical pregnancies were documented. There was a statistically significant difference between the number of clinical pregnancies before and after polypectomy (p < 0.001), as well as between the total number of pregnancies (p < 0.001). Secondary Results: Women with positive outcome were significantly younger and had significantly lower FSH levels (p < 0.007). They also had significantly higher AMH (p < 0.009) and peak estradiol levels (p < 0.013) and yielded more M II oocytes (p < 0.009) and embryos (p < 0.002). Conclusions: Hysteroscopic polypectomy in women with a suspected endometrial polyp using 2D ultrasound and a history of prior failed IVF attempt improves IVF outcomes in terms of the clinical and total number of pregnancies.

Optimizing trigger timing in minimal ovarian stimulation for In Vitro fertilization using machine learning models with random search hyperparameter tuning

Various studies have emphasized the importance of identifying the optimal Trigger Timing (TT) for the trigger shot in In Vitro Fertilization (IVF), which is crucial for the successful maturation and release of oocytes, especially in minimal ovarian stimulation treatments. Despite its significance for the ultimate success of IVF, determining the precise TT remains a complex challenge for physicians due to the involvement of multiple variables. This study aims to enhance TT by developing a machine learning multi-output model that predicts the expected number of retrieved oocytes, mature oocytes (MII), fertilized oocytes (2 PN), and useable blastocysts within a 48-h window after the trigger shot in minimal stimulation cycles. By utilizing this model, physicians can identify patients with possible early, late, or on-time trigger shots. The study found that approximately 27 % of treatments administered the trigger shot on a suboptimal day, but optimizing the TT using the developed Artificial Intelligence (AI) model can potentially increase useable blastocyst production by 46 %. These findings highlight the potential of predictive models as a supplementary tool for optimizing trigger shot timing and improving IVF outcomes, particularly in minimal ovarian stimulation. The experimental results underwent statistical validation, demonstrating the accuracy and performance of the model. Overall, this study emphasizes the value of AI prediction models in enhancing TT and making the IVF process safer and more efficient.

Assessment of immune cells in the uterine fluid at the time of the embryo transfer.

Although endometrial receptivity is a key factor in influencing implantation in both naturally conceived and assisted reproductive technology (ART) cycles, very little is known about the endometrium milieu around the time of implantation. Previous studies have demonstrated the presence of several cytokines in the endometrium that affect implantation. However, there is lacking data about the presence of immune cell subtypes within the endometrium and in the uterine cavity at the time of implantation. This study was approved by the Institutional Review Board (# 225589). The study was designed as a prospective observational cohort study between May 2021 and December 2022 at a single academic-based fertility center. All patients underwent at least one In Vitro Fertilization (IVF) cycle and have frozen embryos. Twenty-four participants were recruited for this study which was conducted during the frozen embryo transfer (FET) cycle regardless of the outcome of previous cycles. Two samples were acquired from each subject, denoted as lower and upper. A trial transfer catheter was introduced under ultrasound guidance into the lower uterine segment. Upon removal, the tip was rinsed in IMDM medium containing 10% FBS (lower uterus). A transfer catheter was then loaded with the embryo that was placed in the upper uterus under ultrasound guidance. The tip of the transfer catheter was rinsed in separate aliquot of the above media (upper uterus). After centrifugation, pelleted cells were stained for the following surface markers: CD45, CD3, CD19, CD4, CD8, gamma delta TCR, CD25, CD127, CD66b, CD14, CD16, CD56 and acquired on Sony SP6800 Spectral Analyzer. Upon staining the pelleted cells, we were able to identify viable leukocytes from samples obtained from both, upper and lower uterus (0.125×106 cells ± SD 0.32), (0.123×106 cells ± SD 0.12), respectively. Among total viable cells, there was no significant difference in both percent and number of CD45+ cells between the upper and lower uterus (9.88% ± 6.98 SD, 13.67% ± 9.79 SD, p=.198) respectively. However, there was significantly higher expression of CD3+ (p=.006), CD19+ (p=.032) and CD14+ (p=.019) cells in samples collected from upper compared to lower uterus. Within all CD3+ cells, we found that gamma delta T cells (GDT) were the major population of T cells in both upper and lower uterus. In contrast, CD8+ T cells were significantly higher in the lower uterus when compared to the upper uterus (p=.009). There was no statistically significant difference in the expression of CD4+ T cells, T regulatory cells (CD4+CD25+CD127-), NK cells (CD56+), neutrophils (CD66b+) and FcγRIII+ cells (CD16+) between upper and lower uterus. We believe the immune milieu at the time of embryo transfer will affect implantation. Understanding the composition of immune cells will guide further research in identifying optimal immune milieus that favor implantation. Comprehensive analysis of endometrium is expected to lead to new diagnostic and therapeutic approaches to improve IVF outcomes.

Assessment of the implantation window and embryonic factor impact to the treatment of recurrent implantation failure (RIF). A prospective study

The aim: to study of the prognostic value of endometrial receptivity and preimplantation genetic diagnosis of embryos, and their influence on the effectiveness of in vitro fertilization (IVF) programs. We also evaluate the importance of this factor in comparison with other potential causes of infertility. Materials and methods: This prospective cohort study included 123 infertile women who underwent in vitro fertilization (IVF) treatment. 93 patients had repeated unsuccessful implantation attempts and were divided into three groups: group 1 - patients who were treated using genetically untested embryos according to a standard fixed stimulation protocol, group 2 - patients who were treated using euploid embryos after preimplantation genetic screening according to standard fixed protocol; group 3 - patients who underwent treatment using euploid embryos after pre-implantation genetic screening and determination of the implantation window with subsequent modification of the stimulation protocol, according to the endometrial examination result. 30 patients had a first attempt at IVF, which was carried out using genetically untested embryos, according to a standard fixed protocol, and made up the control group (CG). Determination of the window of implantation was carried out by triple aspiration biopsy of the endometrium during the luteal phase of the menstrual cycle since the endometrium is most susceptible to implantation during this period. Samples were analyzed using scanning electron microscopy. Based on the results obtained, the endometrial preparation protocol was individualized for the next attempt. preimplantation genetic testing (PGT) of embryos was carried out by the next generation (NGS) method. Statistical analysis was performed using IBM SPSS V25.0 for Windows software. Results: According to the obtained results, patient characteristics, screening rates, IVF cycle characteristics, and the number, quality, and stage of transferred embryos were compared between groups. The rate of clinical pregnancy was 46.7 % among patients of group 1.70 % among patients of group 2, 82.8 % among patients of group 3 and 50.0 % of the control group and statistically significantly different between groups (χ2=10.955, p= 0.012). The rate of live birth was 43.3 % among patients of group 1, 53.3 % among patients of group 2, 72.4 % among patients of group 3 and 43.3 % - of the control group, however, it did not differ statistically significantly between groups (χ2=6,639, р=0,084) Conclusions: The unique window of implantation and the embryonic factor are among the main reasons for multiple failed implantation attempts. Personalization of the endometrial preparation protocol and preimplantation embryo diagnosis are effective methods to improve IVF outcomes

Vaginal microbiota are associated with in vitro fertilization during female infertility.

The vaginal microbiome plays an essential role in the reproductive health of human females. As infertility increases worldwide, understanding the roles that the vaginal microbiome may have in infertility and in vitro fertilization (IVF) treatment outcomes is critical. To determine the vaginal microbiome composition of 1411 individuals (1255 undergoing embryo transplantation) and their associations with reproductive outcomes, clinical and biochemical features are measured, and vaginal samples are 16S rRNA sequenced. Our results suggest that both too high and too low abundance of Lactobacillus is not beneficial for pregnancy; a moderate abundance is more beneficial. A moderate abundance of Lactobacilluscrispatus and Lactobacillus iners (~80%) (with a pregnancy rate of I-B: 54.35% and III-B: 57.73%) is found beneficial for pregnancy outcomes compared with a higher abundance (>90%) of Lactobacillus (I-A: 44.81% and III-A: 51.06%, respectively). The community state type (CST) IV-B (contains a high to moderate relative abundance of Gardnerellavaginalis) shows a similar pregnant ratio (48.09%) with I-A and III-A,and the pregnant women in this CST have a higher abundance of Lactobacillus species. Metagenome analysis of 71 samples shows that nonpregnant women are detected with more antibiotic-resistance genes, and Proteobacteria and Firmicutes are the main hosts. The inherent differences within and between women in different infertility groups suggest that vaginal microbes might be used to detect infertility and potentially improve IVF outcomes.

Sperm DNA Fragmentation: Causes, Effects and Relevance in IVF Outcomes

Sperm DNA fragmentation, characterized by damaged or broken DNA strands within sperm cells, has been recognized as an important factor in male infertility and poor ART outcome. This lecture aims to provide an overview of the causes, effects, and relevance of sperm DNA fragmentation in in vitro fertilization (IVF) outcomes and look into the latest evidences from the literatures. Causes: Sperm DNA fragmentation can result from various factors, including oxidative stress, advanced paternal age, lifestyle factors such as smoking and excessive alcohol consumption, environmental exposures, genotoxic agents, and underlying genetic abnormalities. These factors can induce DNA damage during sperm development, during the transit of sperm through the male reproductive tract or during the storage of the sperm in epididymis. Relevance in IVF Outcomes: The assessment of sperm DNA fragmentation has emerged as a valuable tool in evaluating male infertility and predicting IVF success. Several methods, including the sperm chromatin structure assay (SCSA), terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL), comet assay, and other commercial detection kits, are available to measure sperm DNA fragmentation. Previous small cohort studies have demonstrated that higher levels of sperm DNA fragmentation are associated with lower pregnancy rates, decreased embryo quality, and an increased risk of pregnancy loss in IVF cycles. Therefore, incorporating sperm DNA fragmentation analysis into the routine assessment of male infertility can aid in optimizing treatment strategies and improving IVF outcomes. Recent evidences: Latest large clinical studies showed a limited association to IVF outcome. The DFI remains controversial as a predictive tool. Nonetheless, sperm DNA fragmentation is still believed to be a significant factor affecting male fertility and IVF outcomes. Further research is warranted to explore the mechanisms underlying DNA fragmentation, develop standardized assessment methods, and explore potential interventions to mitigate the adverse effects of sperm DNA damage on IVF success.

#53 : Pregnancy Outcome After Hydrosalpinx Aspiration in In-Vitro Fertilization Cycle

Background and Aims: Hydrosalpinx is a severe manifestation of tubal disease-causing infertility in women. It has an adverse effect on the outcome of in vitro fertilization (IVF), reducing live birth rate (LBR) by 50% compared to other tubal factors. Laparoscopic salpingectomy can effectively improve IVF outcome but is contraindicated in some women. It can also be expensive and is not without complications. Ultrasound-guided hydrosalpinx aspiration is a cheaper, less invasive procedure alternative to salpingectomy. It has been shown to improve clinical and ongoing pregnancy rates compared to patients with no aspiration. However, the published literature on the effectiveness of hydrosalpinx aspiration is equivocal. Method: This retrospective study aimed to evaluate the outcome of minimal stimulation (mild) IVF cycles wherein the women underwent hydrosalpinx aspiration prior to embryo transfer at a private fertility clinic in the Philippines. Results: A total of 98 IVF cycles satisfied the inclusion criteria. The average patient age was 35.22 ± 4.57 and average duration of infertility was 5.87 ± 3.70 years. The biochemical pregnancy rate was 48% and the clinical pregnancy rate was 39.8%. Live birth rate was 35.7%, which is comparable to that of mild IVF patients without hydrosalpinx. There was a negative correlation between patient age and biochemical pregnancy rate, clinical pregnancy rate, and LBR, however, the correlations were statistically insignificant (Pearson correlation, P = 0.081, 0.197, and 0.344, respectively). Binary logistic regression analyses showed that age, infertility type, infertility factor, basal hormone levels, and number of eggs retrieved were not predictive of the success of IVF among the women in the study. Conclusion: Our results show that hydrosalpinx aspiration is a safe and effective option in improving pregnancy and live birth rates in women undergoing mild IVF.

Current Applications and Controversies in Preimplantation Genetic Testing for Aneuploidies (PGT-A) in In Vitro Fertilization.

Preimplantation genetic testing for aneuploidy (PGT-A) has evolved over recent years, including improvements in embryo culture, biopsy, transfer, and genetic testing. The application of new comprehensive chromosome screening analysis has improved the accuracy in determining the chromosomal status of the analyzed sample, but it has brought new challenges such as the management of partial aneuploidies and mosaicisms. For the past two decades, PGT-A has been involved in a controversy regarding its efficiency in improving IVF outcomes, despite its widespread worldwide implementation. Understanding the impact of embryo aneuploidy in IVF (in vitro fertilization) should theoretically allow improving reproductive outcomes. This review of the literature aims to describe the impact of aneuploidy in human reproduction and how PGT-A was introduced to overcome this obstacle in IVF (in vitro fertilization). The article will try to analyze and summarize the evolution of the PGT-A in the recent years, and its current applications and limitations, as well as the controversy it generates. Conflicting published data could indicate the lacking value of a single biopsied sample to determine embryo chromosomal status and/or the lack of standardized methods for embryo culture and management and genetic analysis among other factors. It has to be considered that PGT-A may not be a universal test to improve the reproductive potential in IVF patients, rather each clinic should evaluate the efficacy of PGT-A in their IVF program based on their population, skills, and limitations.

O-065 A multi-center randomized controlled trial of intraovarian injection of platelet rich plasma for women with poor ovarian response

Abstract Study question Does intraovarian injection of autologous platelet-rich plasma (PRP) improve oocyte yield in young patients with poor ovarian response (POR)? Summary answer Intra-ovarian PRP injection did not result in increased number of oocytes retrieved or improvement of any other relevant measured outcomes. What is known already POR is one of the major hurdles to overcome in patients undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). Several methods have recently emerged to attempt follicular reactivation in these patients with a poor prognosis. One of them, the intra-ovarian injection of autologous PRP, demonstrated promising results in retrospective and prospective cohort studies. Study design, size, duration This was a multi-center, randomized controlled trial (RCT) to evaluate the efficacy of PRP in improving IVF outcomes in women with POR. Patients who met inclusion criteria (&amp;lt;38 years, two or more prior cycles with &amp;lt; 3 oocytes retrieved, and without single gene disorders, prior ovarian surgery, endometriomas, BMI &amp;gt;35, or severe male factor infertility) were randomized to either receive an autologous intra-ovarian PRP injection (PRP group) or no intervention (control group) prior to COH. Participants/materials, setting, methods A total of 83 patients were randomized to PRP group (n = 41) or control group (n = 42). After treatment, patients underwent COH, oocyte retrieval, intracytoplasmic sperm injection (ICSI), preimplantation genetic testing for aneuploidy (PGT-A), and single frozen euploid embryo transfer. Number of MII oocytes obtained was the primary outcome. Secondary outcomes included ovarian reserve tests (antral follicle count [AFC] and anti-Mullerian hormone [AMH]), blastocyst and euploid blastocyst yields, and sustained implantation. Main results and the role of chance No significant differences were observed in number of MII oocytes retrieved (3.1 ± 3.3 vs 2.8 ± 2.4 in PRP vs control, respectively; p = 0.9), blastocysts (1.3 ± 2.1 vs 1.0 ± 1.3, p = 0.8), or euploid blastocysts (0.9 ± 1.6 vs 0.8 ± 1.1; p = 0.5) per cycle. Similarly, no differences were observed in the likelihood of obtaining at least one euploid blastocyst (37 vs 45%, p = 0.4; relative risk [RR], 95% confidence interval [CI] 0.9, 0.6-1.2) or the rate of sustained implantation (29 vs 31%, p = 0.9; RR 1.0, 0.7-1.3). Pre- and post-treatment AFC in both groups (PRP: from 5.2 ± 3.2 to 7.9 ± 4.5, p &amp;lt; 0.0001; control: from 5.6 ± 3.3 to 6.8 ± 4.8, p = 0.02) were statistically significantly different, whereas AMH was different in the PRP group only (PRP: 0.73 ± 0.46 to 0.99 ± 0.98, p = 0.02; control: 0.70 ± 0.59 to 0.73 ± 0.57, p = 0.59). Limitations, reasons for caution Oocyte retrieval took place the cycle immediately following treatment; therefore, we could not evaluate long-term effects of PRP on ovarian reserve or response. Lack of a placebo group would limit interpretability had a difference between groups been observed. The effect of PRP on patients &amp;gt;38 years old was not studied. Wider implications of the findings Intra-ovarian PRP injection did not improve reproductive outcomes by any relevant measure. Our findings do not support wide utilization of PRP to improve IVF outcomes in women with POR. Trial registration number NCT04163640

P-112 Outcome of laser collapsing blastocyst on clinical pregnancy rate

Abstract Study question To determine clinical pregnancy rate after laser-induced collapse of blastocyst before cryopreservation Summary answer Laser collapse of the blastocyst before cryopreservation increases clinical pregnancy rate regardless of assigned embryo quality group compared to the blastocysts vitrified without laser collapsing What is known already Vitrification has been associated with the certain risks to the structure of an embryo due to intracellular ice crystal formation and impaired shrinkage rate. Blastocyst is a large fluid filled cavity requiring additional preparation to reduce the potential risks of damage. Collapsing a blastocyst has been shown to improve survival rate of an embryo following thawing and improve in vitro fertilization (IVF) outcomes including clinical pregnancy and live birth rates Study design, size, duration . It is a single-centre retrospective study conducted at a private clinic in Kazakhstan. We analyzed the outcome of IVF treatment following cryopreservation and subsequent frozen embryo transfer of 2959 blastocysts from 1642 women during January 2019-December 2021 Participants/materials, setting, methods The data on blastocyst cryopreservation technique and IVF treatment outcome was collected from the clinic’s electronic database. The blastocysts were divided into two groups based on performed laser collapsing prior to vitrification and further stratified by embryo quality group including excellent, good and fair groups. The primary outcome was clinical pregnancy rate. Laser-assisted collapsing procedure was initiated in August 2020 at the clinic. Thus, all previously vitrified blastocysts served as control group for the study Main results and the role of chance 1556 blastocysts from 777 women vitrified following laser collapsing and 1403 blastocysts from 865 women without prior laser collapsing were analyzed in the study. Stratification of blastocysts by the embryo quality were comparable between two groups with more than half of embryos defined as excellent embryo quality group, 54% in collapsed group and 52% in non-collapsed group (p &amp;gt; 0.05). Most blastocysts were from women under the age of 35 years old in both study groups. 1 in 4 collapsed blastocysts of the excellent embryo quality group were from women older than 35 years old, whereas non-collapsed group of blastocysts was evenly distributed between embryo quality groups (p &amp;lt; 0.05). Clinical pregnancy rate was the highest in the excellent embryo quality group with prior laser collapsing compared to the group without collapsing, 53% vs 43% respectively (p &amp;lt; 0.001). Overall, collapsed blastocysts showed higher rate of clinical pregnancy rate in all three embryo quality groups. Despite of changes associated with the COVID-19 pandemic IVF outcomes had not been affected but improved with the use of laser collapsing prior to embryo vitrification Limitations, reasons for caution Retrospective design of the study did not provide full baseline information on potential confounders including paternal characteristics and concurrent conditions. Moreover, the study period included the COVID-19 pandemic start which could have affected treatment outcomes Wider implications of the findings Collapsing the blastocyst embryo before vitrification should become a routine procedure to improve treatment outcomes of frozen embryo transfer irrespective of woman’s age and embryo’s quality Trial registration number not applicable

A Randomized Controlled Intervention Trial with Danazol to Improve Telomeric and Fertility Parameters in Women with Diminished Ovarian Reserve: A Pilot Study.

Most women who are treated at in vitro fertilization (IVF) clinics have trouble conceiving due to ovarian failure (OF), which seems to be associated to short telomeres and reduced or absent telomerase activity in their granulosa cells. Indeed, telomere pathways are involved in organ dysfunction. However, sexual steroids can stimulate the expression of the telomerase gene and have been successfully used to prevent telomere attrition. Thus, a strategy to improve IVF outcomes in women with OF could be telomerase reactivation using sexual steroids. We conducted a double-blind, placebo-controlled study. Patients with diminished ovarian reserve were randomized to Danazol or placebo for 3 months. We included patients with normal ovarian reserve in the study as untreated controls. Patients and controls underwent several ovarian stimulations (OSs). Telomere and IVF parameters were assessed. We found that the mean telomere length in blood and the percentage of short and long telomeres were similar throughout the 3 months of treatment with Danazol. Remarkably, while the number of cells with one telomeric repeat-containing RNA (TERRA) focus decreased (p = 0.04) after the first month of Danazol treatment, the number of cells with 2 to 4 TERRA foci increased (p = 0.02). Regarding fertility, no differences were found in the antral follicle count. Interestingly, in OS performed after the trial, all Danazol-treated patients had a better MII oocyte rate compared to OS performed before the pilot study.EudraCT number: 2018-004400-19. Danazol treatment seemed to affect telomere maintenance, since both the number of TERRA foci and the ratio of MII oocytes changed. However, further research is needed to confirm these results.

Local glucose elevation activates pyroptosis via NLRP3 inflammasome in ovarian granulosa cells of overweight patients.

Overweight, with an increasing prevalence worldwide, significantly impairs the clinical outcomes following in vitro fertilization (IVF). Hyperglycemia, hyperlipidemia, and metabolic disorders are always accompanied by the majority of overweight patients. The association between granulosa cell function and metabolic alterations in follicular fluid including lipids, proteins, and growth factors has been extensively documented. However, the effects of higher glucose level on ovarian granulosa cells (GCs), remain largely unknown. In this study, we identified that overweight women had elevated follicular glucose level which profoundly activated NLRP3 inflammasome and pyroptosis. An in vitro correlation between follicular high glucose, NLRP3 inflammasome and pyroptosis was also established. More importantly, in granulosa cells of overweight patients, the activation of the NLRP3 inflammasome and pyroptosis induced by high glucose was involved in the dysregulation of estradiol synthesis. Our study may provide new options to interpretate and improve IVF outcomes in overweight women.

Is the long-acting gonadotropin-releasing hormone agonist longprotocol better for patients with endometriosis undergoing IVF?

To investigate the effect of the long-acting gonadotropin-releasing hormone agonist (GnRHa) long protocol on in vitro fertilization (IVF) outcomes of patients with endometriosis (EMs). This retrospective cohort study was carried out from July 1, 2016 to June 30, 2021. In all, 798 patients with EMs who underwent first IVF were enrolled. The patients were classified by the ovarian stimulation protocols. The clinical outcomes of IVF were compared in each group. Those EMs patients who received the long-acting GnRHa long protocol had significantly higher clinical pregnancy rate (72.00%, 60.70% and 50.90%, respectively; P= 0.047 and 0.010) and implantation rate (51.0%, 44.6%, and 38.7%, respectively; P= 0.006 and <0.001) compared with the short-acting GnRHa long protocol and the GnRH antagonist protocol. Live birth rate was also significantly higher than the GnRH antagonist protocol (60.10% vs. 40.0%, P= 0.032), but not statistically different from the short-acting GnRHa (60.10% vs. 53.80%, P= 0.443). In addition, they also had significantly higher duration of stimulation, total dose of gonadotropin, and number of high-quality embryos transferred compared with other groups (P< 0.001). The long-acting GnRHa long protocol could improve IVF outcomes of patients with EMs compared with the short-acting GnRHa long protocol and the GnRH antagonist protocol.

Transfer of the fittest: using preimplantation genetic testing for aneuploidy to select embryo(s) most likely to lead to live birth

Preimplantation genetic testing for aneuploidy (PGT-A) was developed to identify euploid embryos from a cohort of embryos with unknown ploidy produced during an invitro fertilization (IVF) cycle. In recent years, the ability of PGT-A to improve IVF outcomes has come into question. The goal of this review was to summarize the major randomized controlled trials (RCTs) and nonselection studies evaluating the benefit of PGT-A to improve the live birth rates (LBRs). We argue that the LBR per transfer is more relevant to the individual patient than the cumulative LBR as a means to minimize the burden of IVF by reducing futile transfers, pregnancy losses, and ongoing aneuploidy. The early RCTs demonstrate improved implantation rates and LBRs with PGT-A for embryo selection vs. traditional morphology. However, these studies are limited by the small sample size and a bias toward good-prognosis patients. Further studies using next-generation sequencing found more variable results but did confirm an improvement in the LBRs per transfer in an older population with a higher baseline risk of aneuploidy. The largest RCT to date showed similar cumulative LBRs in the PGT-A and control groups after biopsy and sequential transfer of up to 3 blastocysts with a significant reduction in the cumulative clinical pregnancy loss rate in the PGT-A group. Nonselection studies evaluating pregnancy outcomes after transfer of euploid vs. aneuploid embryos demonstrate near-perfect negative predictive value for an aneuploid result to predict live birth. Putative mosaic embryos had similar LBRs compared with euploid embryos. The available RCTs and nonselection studies support the practice of using PGT-A to identify euploid embryos for transfer, especially in an older population, while simultaneously selecting against aneuploid embryos, without negative impact on the total reproductive potential of the cycle.

P-629 A prospective, randomized, placebo-controlled pilot trial investigating telomere length dynamics and ovarian function in patients with diminished ovarian reserve (DOR) after danazol treatment

Abstract Study question Could the administration of a synthetic steroid -danazol- for 3 months reactivate telomerase activity and lengthen telomeres in patients with DOR? Summary answer Danazol treatment resulted in an increase of the mean leukocyte telomeric length (TL) but did not improve fertility results after the nearest ovarian stimulation (OS). What is known already A primary molecular cause of aging is telomere attrition, given that the accumulation of critically short telomeres leads organ dysfunction. Telomere shortening can be counteracted by the telomerase enzyme, which is responsible for adding deoxyribonucleotides to the linear ends of chromosomes. In patients with DOR, TL in granulosa cells (GCs) is short and telomerase activity is absent, compromising their proliferative capacity. Previuos studies have shown that estrogens can activate telomerase gene transcription through estrogen-responsive elements located in the promoter of the telomerase gene. Therefore, telomere attrition could be reduced in patients with DOR upon telomerase reactivation. Study design, size, duration This blinded, placebo-controlled pilot trial was carried out at IVIRMA Madrid clinic between February 2020 and February 2022. The study included 12 patients with DOR (AMH&amp;lt;2ng/ml) which were randomized to danazol or placebo. Patients had 12 visits for health supervision and in vitro fertilization (IVF) treatment. Both groups received danazol (400mg per day) or placebo for 3 months. As control group, 7 patients with normal ovarian reserve (AMH≥2ng/ml) were included. Participants/materials, setting, methods In both groups, blood samples were collected prior (PT) and after the treatment (AT). Luteinized GCs and blood samples were obtained during oocyte retrieval, after OS. Leukocytes were purified in Ficoll gradients and then, evaluated for TL and TRF1 levels by Quantitative Fluorescent in Situ Hybridization followed by image acquisition using high resolution confocal microscope. IVF parameters were assessed according to IVIRMA standard protocols. Main results and the role of chance A total of 19 women were recruited: 5 in the placebo group (39.8±2.7 years), 7 in Danazol group (41.0±3.0 years) and 7 in control group (39.5±3.0 years). The Danazol group showed a trend to higher mean TL in leukocytes after the treatment (108.2±37.4 versus 99±13.2 a.u., AT vs PT). Interestingly, the percentage of long telomeres also increased with treatment (15.5±0.0 versus 10.8±8.1 a.u., AT vs PT). In addition, there was a decrease of the percentage of critically short telomeres (15.7±0.0 versus 20.8±9.9 a.u. AT vs PT). In the placebo group, mean leukocyte TL and the percentage of long and short telomeres remained constant. These findings suggest that telomeres lengthen after Danazol treatment. The protein levels of TRF1, a shelterin involved in telomere protection, were similar in both Danazol and placebo groups. Regarding fertility, there were no differences in the number of antral follicles, total and MII oocytes, fertilization rate and blastocyst formation. The control group had, as expected, signficantly higher mean follicle number after OS when compared with Danazol (16.0±1.4 and 0.7±0.9, P &amp;lt; 0.0001) and placebo (16.0±1.4 and 5.0±3.8, p = 0.0127) groups, and higher numbers of total oocytes retrieved (p = 0.006; control and Danazol, and p = 0.007; control and placebo). Limitations, reasons for caution Firstly, the number of patients included in the study is limited due to strict inclusion/exclusion criteria. Secondly, because follicle growth can take up to 150 days, danazol effects could be noticeable at later OS and not only in the first OS, which was the only one studied here. Wider implications of the findings Since telomeres seem to elongate in blood samples after Danazol treatment in DOR patients, we could hypothesize that this may have a positive impact for general health, including the ovaries. Thus, danazol therapy might help improving IVF outcomes, perhaps at later OS. Trial registration number NCT04058678

Use of pentoxifylline during ovarian stimulation to improve oocyte and embryo quality: A retrospective study.

ObjectiveThis study aims to investigate if the administration of pentoxifylline during ovarian stimulation have an impact on the quantity and quality of oocytes recovered for in vitro fertilization (IVF), the quality of zygotes and embryos resulting from IVF, the serum estradiol and progesterone concentrations, the endometrial thickness and pregnancy outcomes after embryo transfer. MethodsRetrospective crossover study including 18 women with recurrent implantation failur, defined as having a history of at least two consecutive cycles of implantation failure and 25-45 years of age at the time of embryo transfer. Pentoxifylline was added during their following IVF attempt aiming to improve endometrial function. Oocyte, zygote and embryo quality, serum estradiol and progesterone concentrations, endometrial thickness and IVF outcomes were compared between the two sequential attempts. ResultsCompared to the first attempt, the outcomes of the second attempt, with the addition of pentoxifylline administration, resulted in higher numbers of total and mature oocytes, better oocyte, zygote and embryo quality, higher serum estradiol concentrations, thicker endometrium and better clinical IVF outcomes. ConclusionsThe administration of pentoxifylline during ovarian stimulation seems to improve IVF outcomes, not only by the generating a thicker endometrial lining, but also by producing a higher number of oocytes, with better oocyte, zygote and embryo quality, and higher serum estradiol concentrations.

DHEA restores mitochondrial dynamics of cumulus cells by regulating PGAM5 expression in poor ovarian responders

ObjectivePoor ovarian responder (POR) poses a significant challenge for in vitro fertilization (IVF). Previous studies have suggested that dehydroepiandrosterone (DHEA) may improve IVF outcomes in POR. The current study attempts to investigate the clinical benefits of DHEA in POR and the possible mechanism of DHEA on cumulus cells (CCs). Materials and methodsA total of 60 women who underwent IVF treatment participated, including 22 normal ovarian responders (NORs) and 38 PORs. PORs were assigned to receive DHEA supplementation (n = 18) or not (n = 20) before IVF cycles. For all patients, CCs were obtained after oocyte retrieval. In the CCs, mRNA expression of mitochondrial dynamics relataed genes were measured. ResultsSupplementation of DHEA in POR reduced mitochondrial fission in CCs and decreased the expression of PGAM5 in CCs. ConclusionThe benefit of DHEA supplementation on IVF outcomes in POR is significant, and this effect may be mediated in part through improved mitochondrial dynamics in CC.