Improved clinical outcomes after non-invasive oocyte selection and Day 3 eSET in ICSI patients.

BACKGROUND: Non-invasive oocyte quality scoring, based on cumulus gene expression analysis, in combination with morphology scoring, can increase the clinical pregnancy (CPR) and live birth rates (LBR) in Day 3 eSET (elective single embryo transfer) ICSI patients. This was first investigated in a pilot study and is now confirmed in a large patient cohort of 633 patients. It was investigated whether CPR, LBR and time-to-pregnancy could be improved by analyzing the gene expression profile of three predictive genes in the cumulus cells, compared to patients with morphology-based embryo selection only. METHODS: A large interventional, non-randomized, assessor-blinded cohort study with 633 ICSI patients was conducted in a tertiary fertility center. Non-PCOS patients, 22-39 years old, with good ovarian reserve, were stimulated with HP-hMG using a GnRH antagonist protocol and planned for fresh Day 3 eSET. The cumulus cells from individually denuded oocytes were ranked by a lab-developed cumulus cell test: qRT-PCR for three predictive genes (CAMK1D, EFNB2 and SASH1) and two control genes (UBC, B2M). The embryo selected for transfer was highest ranked from the pool of morphologically transferable Day 3 embryos. Patients in the control (n = 520) and experimental arm (n = 113) were compared for clinical pregnancy and live birth, using a weighted generalized linear model, and time-to-pregnancy using Kaplan-Meier curves. RESULTS: The CPR was 61% in the experimental arm (n = 113) vs 29% in the control arm (n = 520, p < 0.0001). The LBR in the experimental arm (50%) was significantly higher than in the control arm (27%,p < 0.0001). Time-to-pregnancy was significantly shortened by 3 transfer cycles independent of the number of embryos available on Day 3 (Kaplan-Meier, p < 0.0001). Cumulus cell tested patients < 35 years (n = 65) or ≥ 35 years (n = 48) had a CPR of 62 and 60% respectively (ns). For cumulus cell tested patients with 2, 3-4, or > 4 transferable embryos, the CPR was 66, 52, and 67% (ns) respectively, and thus independent of the number of transferable embryos on Day 3. CONCLUSIONS: This study provides further evidence of the clinical usefulness of the non-invasive cumulus cell test over time in a larger patient cohort. TRIAL REGISTRATION: Clinicaltrials.gov, NCT03659786 / NCT02962466 (Registered 6Sep2018/11Nov2016, retrospectively registered.

IVF in women aged 43 years and older: a 20-year experience.

RESEARCH QUESTION: What are the reproductive outcomes of women aged 43 years and older undergoing IVF and intracytoplasmic sperm injection (ICSI) treatment using their own eggs. DESIGN: Retrospective study of 833 woman aged 43 years or older undergoing their first IVF and ICSI cycle using autologous oocytes at a tertiary referral hospital between January 1995 and December 2019. Live birth rate (LBR) after 24 weeks' gestation was the primary outcome. RESULTS: Ninety-five out of 833 (11.4%) had a positive HCG, whereas 59 (62.1% per positive HCG) had a miscarriage before 12 weeks' gestation and 36 (4.3%) live births were achieved. Analysis by age showed that the number of cumulus-oocyte complexes retrieved was significantly different between the four age groups: 43 years (5 [3-9]); 44 years (5 [2-7]); 45 years (3 [2-8)]); ≥45 years (2.5 [2-6]); P < 0.01; the number of metaphase II oocytes, however, was similar. Positive HCG rates remained low: 43 years (78/580 [13.4%]); 44 years (14/192 [7.3%]); 45 years (1/39 [2.6%]; and ≥46 years (2/22 [9.1%]); P = 0.03, as did LBR: 43 years (28 [4.8%]); 44 (7 [3.6%]); 45 years (0 [0%]); and ≥46 years (1 [4.5%]); P = 0.5. Multivariate regression analysis revealed that only number of metaphase II was significantly associated with LBR, when age was considered as a continuous (OR 1.08, 96% CI 1.004 to 1.16) or categorical variable (OR 1.08, 95% CI 1.005 to 1.16). CONCLUSION: The chances of achieving a live birth in patients aged 43 years and older undergoing IVF/ICSI with their own gametes are low, even in cases of patients with a relatively 'normal' ovarian reserve for their age.

Is ovarian response associated with adverse perinatal outcomes in GnRH antagonist IVF/ICSI cycles?

RESEARCH QUESTION: Is there an association between ovarian response and perinatal outcomes? DESIGN: A retrospective, single-centre cohort study including all women undergoing their first ovarian stimulation cycle in a gonadotrophin releasing hormone antagonist protocol, with a fresh embryo transfer that resulted in a singleton live birth from January 2009 to December 2015. Patients were categorized into four groups according to the number of oocytes retrieved: one to three (category 1), four to nine (category 2), 10-15 (category 3), or over 15 oocytes (category 4). RESULTS: The overall number of patients analysed was 964. No relevant statistical difference was found among neonatal outcomes across the four ovarian response categories. Neonatal weight (in grams) was comparable between all groups (3222 ± 607 versus 3254 ± 537 versus 3235 ± 575 versus 3200 ± 622; P = 0.85, in categories 1, 2, 3 and 4, respectively). No statistically significant differences were found among the ovarian response categories for birth weight z-scores (taking into account neonatal sex and delivery term). The incidence of pre-term birth and low birth weight was comparable across the different ovarian response groups (P = 0.127 and P = 0.19, respectively). Finally, the occurrence of adverse obstetric outcomes did not differ among the ovarian response categories. Multivariate regression analysis revealed that the number of oocytes was not associated with neonatal birth weight. CONCLUSIONS: No association was found between ovarian response and adverse perinatal outcomes in antagonist IVF and intracytoplasmic sperm injection cycles. Future, larger scale and prospectively designed investigations are needed to validate these results.

The effect of cigarette smoking on the semen parameters of infertile men.

INTRODUCTION: 36.9% of men worldwide use tobacco. Previous studies suggest a negative effect of cigarette smoking on semen quality, but the results are contradictory. We have studied the effects of smoking on the semen characteristics such as sperm concentration, semen volume, sperm motility, sperm vitality and sperm morphology in a large group of infertile men. METHODS: This retrospective study was conducted on a total of 5146 infertile men with at least one year of idiopathic infertility, who admitted to the Centre for Reproductive medicine (CRG) at the Brussels University Hospital, Belgium between 2010 and 2017. The smokers were classified as mild (1-10 cigarettes/d), moderate (11-20 cigarettes/d) or heavy smokers (> 20 cigarettes/d). Semen analysis was performed for all patients. Statistical analysis was performed using the R software package and t-test or Mann-Whitney U tests were used, group comparisons were performed using ANOVA, ANCOVA or Kruskal-Wallis tests as appropriate. A p-value <0.05 was considered as statistically significant. RESULTS: Comparing the semen parameters in the two global groups showed that smoking had a significant decrease in semen volume (p=0.04074) and sperm concentration (p=0.029). ANOVA testing on the different smoking groups versus non-smoking group showed a significant decrease in sperm concentration (p=0.0364). After adjusting for the confounders, age and testosterone, ANCOVA testing showed significant effect on the sperm concentration (p=0.03871) in smokers versus non-smokers. No significant correlation was detected between the other semen characteristics. CONCLUSION: We concluded that smoking had a significant and independent effect on the sperm concentration in a semen analysis. Other parameters, like semen volume, sperm motility, sperm vitality and sperm morphology were not influenced by smoking.

Does the type of GnRH analogue used, affect live birth rates in women with endometriosis undergoing IVF/ICSI treatment, according to the rAFS stage?

Since the introduction of gonadotropin-releasing hormone (GnRH) antagonists, an extensive amount of literature investigating the role of the downregulation protocols on pregnancy outcomes has been published. However, these studies were mainly performed in the general infertile population where patients with endometriosis were often excluded or underrepresented. This study is a large retrospective cohort study including 386 endometriosis patients undergoing IVF/ICSI, who had been previously classified according to the rAFS system. Patients were stimulated either a long GnRH agonist or GnRH antagonist protocol. Depending on endometriosis stage, patients were divided into two groups: endometriosis stage I-II and endometriosis stage III-IV. Each group was subdivided, based on the type GnRH analog used. When comparing the GnRH agonist and antagonist groups, patients with endometriosis stage I-II, had a tendency toward higher ß-hCG positive, clinical pregnancy, and live birth rates (42.8% vs. 26.7%; p = .07) in favor of GnRH agonist use. In endometriosis stage III-IV, no differences were observed between agonist and antagonist cycle in any of the pregnancy outcomes. Multivariate regression analysis did not reveal any significant predictor of live birth after adjusting for relevant confounders. Based on our findings, the chance to have a liveborn in endometriosis population seems not to be affected by the type of GnRH analog used, at least in advanced stages. Findings from stage I-II endometriosis cases merit consideration and further evaluation in a larger sample size is warranted.

Cumulative delivery rates after ICSI with donor spermatozoa in different age groups.

This retrospective cohort study followed a total of 364 women from their first fresh, donor intracytoplasmic sperm injection (ICSI) cycle through to up to six ICSI cycles. All patients started their treatment between January 2003 and December 2007. Live delivery after 25 weeks of gestation was the main outcome measure. The overall crude cumulative delivery rate (CDR) after six cycles was 66% while the overall expected CDR was 90%. In women aged 38-39 years, the crude and expected CDR after six cycles were 54% and 82%, respectively. In women aged 30-37 years, the crude and expected CDR after six cycles were 66% and 91%, respectively. In women aged 20-29 years the crude and expected CDR after six cycles were 81% and 93%, respectively. No significant difference was found between the CDR of patients who had a primary ICSI treatment (no previous intrauterine insemination) and patients who had previous intrauterine insemination. This study corroborates the impact of age on ICSI with donor spermatozoa.

Cumulative success rates following mild IVF in unselected infertile patients: a 3-year, single-centre cohort study.

A 3-year, retrospective, single-centre cohort study was conducted in a private infertility centre to determine cumulative live birth rates (LBR) per scheduled oocyte retrieval following minimal ovarian stimulation/natural-cycle IVF in unselected infertile patients. A total of 727 consecutive infertile patients were analysed who underwent 2876 (median 4) cycles with scheduled oocyte retrieval from November 2008 to December 2011. Natural-cycle IVF or clomiphene-based minimal ovarian stimulation was coupled with single-embryo transfer and increased use of delayed vitrified-warmed blastocyst transfer. Main outcome measures were crude and expected age-specific cumulative LBR per scheduled oocyte retrieval. Crude cumulative LBR were 65%, 60%, 39%, 15% and 5% in patients aged 26-34, 35-37, 38-40, 41-42 and 43-44 years, respectively. No live births occurred in patients aged ⩾ 45 years. Dropout rates per cycle were 13-25%. Success rates gradually reached a plateau, with few additional live births after six cycles. Most of the expected success rate was reached within 6 months with almost maximal rates within 15 months of the first oocyte retrieval. Acceptable cumulative LBR are reached with an exclusive minimal ovarian stimulation/single-embryo transfer policy especially in patients aged <38 years but also in intermediate aged patients (38-40 years).

Non-invasive cumulus cell analysis can be applied for oocyte ranking and is useful for countries with legal restrictions on embryo generation or freezing.

RESEARCH QUESTION: Can a strategy for scoring oocyte quality, based on cumulus cell (CC) gene expression, prioritize oocytes with the highest implantation potential, while limiting the number of embryos to be processed in culture and the number of supernumerary embryos to be vitrified? DESIGN: An interventional, blinded, prospective cohort study was retrospectively analyzed. In the original study, patients underwent a fresh Day3 single embryo transfer with embryos ranked based on morphology and CC gene expression (Aurora Test). The additional ranking of the embryos with the Aurora Test resulted in significant higher clinical pregnancy and live birth rates. Now it is investigated if the Aurora Test ranking could be applied to select oocytes. The effect of an Aurora Test based restriction to 2 and 3 2PN or MII oocytes on clinical pregnancy and other outcomes, was analyzed in two subsets of patients with all 2PN (n = 83) or all MII oocytes (n = 45) ranked. RESULTS: Considering only the top three ranked 2PN oocytes, 95% of the patients would have received a fresh SET on Day3 resulting in 65% clinical pregnancies. This was not different from the pregnancy rate obtained in a strategy using all oocytes but significantly reduced the need for vitrification of supernumerary embryos by 3-fold. Considering only top-ranked MII oocytes gave similar results. CONCLUSIONS: In countries with legal restrictions on freezing of embryos, gene expression of CC can be used for the selective processing of oocytes and would thus decrease the twin pregnancy rate and workload, especially for embryo morphology scoring and transfers as the handling and processing of lower competence oocytes is prevented, while improving the ART outcome.

Assisted reproduction using donor spermatozoa in women aged 40 and above: the high road or the low road?

The effect of age on outcome is one of the most intriguing areas in the assisted reproduction field. In older patients using donor spermatozoa to reproduce, it remains undefined as to which is the treatment of choice: intrauterine insemination (IUI) or IVF/intracytoplasmic sperm injection (ICSI). Since life-table analysis provides data that are easy to use for patient counselling, this study analysed cumulative delivery rates (CDR) in patients using donor spermatozoa undergoing either primarily IUI or IVF/ICSI and patients who eventually switched from IUI to IVF/ICSI. Crude and expected CDR after six IUI cycles and three primary ICSI cycles (no previous IUI) were similar in both groups (24% versus 26% and 29% versus 35%, respectively). Since time-to pregnancy is an important factor in these older patients, ICSI treatment is advised to be started immediately, since a single cycle of ICSI will achieve the same success rate as a much longer period with at least six IUI cycles. If patients switch to ICSI after failed IUI, this only adds marginal benefit in CDR. Nearly all deliveries in the primary ICSI group were achieved in the first cycle.

Assisted reproduction counseling in women aged 40 and above: a cohort study.

PURPOSE: Fertility treatment in women aged ≥40 year old remains difficult and controversial. All available studies in older women report results of one specific method of ART, i.e. IUI, IVF/ICSI or oocyte donation, and success rates are always published per attempt but never per patient. Randomized studies are not available because of the obvious heterogeneity in patient populations and treatment options. This prospective observational study aimed at analyzing the outcome in a consecutive cohort of patients above 40 undergoing various methods of ART. METHODS: A total number of 909 women older than 40 attended our fertility centre during a 3 years period. A flowchart showing the consecutive ART treatments with their respective outcome was constructed. Any delivery after 22 weeks gestation (or 500 g.) was taken as primary endpoint. Crude cumulative delivery rates (CDRs) and binomial exact 95 % confidence limits (95 % CLs) were calculated for each group of interest. RESULTS: ART treatment could be proposed to 737 patients (81 %) and eventually 585 patients (64 %) started ART treatment: 111 patients started IUI, 439 patients started IVF/ICSI and 35 patients started oocyte donation as a primary approach ART. Ten patients got pregnant spontaneously and delivered before starting any treatment. In the 909 patients consulting for infertility, 111 deliveries were achieved after ART, i.e. a crude CDR of 12.2 % (95 % CL 10.1 % to 14.5 %). CONCLUSION: Only 10 % of patients aged 40 and above could achieve delivery of their genetically-own child, while 1 % conceived spontaneously. More than one third of patients consulting never started any treatment for different reasons, i.e. anticipated poor prognosis, financial restrictions, illness or spontaneous pregnancy.

The experience of women who delivered during the first wave of COVID-19 pandemic in Belgium: a retrospective study.

BACKGROUND: The lockdown caused by the COVID-19 pandemic has imposed some restrictions on hospital activities, requiring medical staff to find efficient alternatives to ensure adequate medical care for patients. OBJECTIVE: This study aimed to investigate the experience of pregnant women who delivered during the first wave of COVID-19, and to evaluate the impact of COVID-19-related restrictions. STUDY DESIGN: This was a retrospective multicenter study. All pregnant women who delivered a live infant between March 20, 2020 and June 20, 2020 were evaluated using a 35-item survey at 1 year following delivery. Each patient was contacted via 3 modalities. Patients who reported that their prenatal follow-up was interrupted were compared with those who reported that their prenatal follow-up was unchanged. Among 1096 patients who delivered a live infant across the 3 participating centers during the study period, 389 responses were needed for an estimated margin of error of 4%. RESULTS: A total of 469 of 1096 (42.8%) patients answered the survey, of whom 151 (32.2%) reported that the follow-up of their pregnancy was interrupted (exposed group) and 318 (67.8%) reported that their follow-up was maintained as normal (unexposed group). The rate of presentation to the emergency department was higher in the exposed group than in the unexposed group (P=.001). The level of dissatisfaction was also higher in the exposed group, and patients in this group would have postponed their pregnancy if they had known about the pandemic in advance (P<.001 and P=.001, respectively). CONCLUSION: Interruption and modification of antenatal follow-up in pregnant women is associated with patient dissatisfaction and increased presentation to the emergency department.

Undetectable viral RNA in follicular fluid, cumulus cells, and endometrial tissue samples in SARS-CoV-2-positive women.

OBJECTIVE: To study the presence of viral RNA in the follicular fluid, cumulus cells, and endometrial tissue samples in SARS-CoV-2-positive women undergoing assisted reproductive technology (ART). DESIGN: Prospective, single-center, observational study. SETTING: Tertiary hospital. PATIENT(S): A total of 16 patients undergoing transvaginal oocyte retrieval who had a positive SARS-CoV-2 RNA test <48 hours before the procedure. All patients underwent the retrieval between September 2020 and June 2021 and used in vitro fertilization or intracytoplasmic sperm injection. All embryos were vitrified to avoid conception during SARS-CoV-2 infection. INTERVENTION(S): Follicular fluid aspirated during oocyte retrieval, cumulus cells, and endometrial samples were analyzed for SARS-CoV-2 RNA using the RealStar SARS-CoV-2 RT-PCR-Kit1.0. MAIN OUTCOME MEASURE(S): The primary outcome parameter was the detection of viral RNA in the follicular fluid, cumulus cells, and endometrial cells. Fertilization rate, embryo developmental potential, and clinical outcome after frozen embryo transfer were secondary outcome parameters. RESULT(S): Samples from 16 patients were analyzed. Cycle threshold values of <40 were considered positive. All samples were negative for SARS-CoV-2 viral RNA. No inflammatory lesions of the endometrium were identified histologically. Fertilization rate, embryo development, and clinical outcomes after embryo transfer were reassuring. CONCLUSION(S): In women infected with SARS-CoV-2 who underwent ART, viral RNA was undetectable in the follicular fluid, cumulus cells, and endometrium. Caution is warranted in view of the small sample size, and the risk of SARS-CoV-2 affecting the embryo via ART cannot be ruled out. Adequate counseling of women and couples undergoing ART is crucial in parallel with further research on the effect of exposure of the early human embryo to SARS-CoV-2. CLINICAL TRIAL REGISTRATION NUMBER: NCT04425317.

Continuous ropivacaine subfascial wound infusion after cesarean delivery in pain management: A prospective randomized controlled double-blind study.

OBJECTIVE: The aim of this study is to analyze post-cesarean morphine consumption using continuous ropivacaine subfascial wound infusion. METHODS: After standardized spinal anesthesia (0.5% hyperbaric bupivacaine 8-10 mg combined with sufentanil 2-2.5 µg), women undergoing cesarean section (n = 69) were randomly allocated to receive either ropivacaine 0.2% (n = 35) or NaCl 0.9% (n = 34) infused through a subfascial wound catheter during 48 h in a multimodal analgesic approach. As primary outcome, opioid use by intravenous patient-controlled analgesia was analyzed. Secondary outcomes were intensity of pain on visual analog scale at rest and at mobilization, postoperative nausea/vomiting, pruritus and time of first ambulation. Independent t test or Mann-Whitney U test, and Pearson's χ2 test or Fisher's exact test were used as appropriate. RESULTS: Morphine consumption was significantly lower in the ropivacaine group (21.52 ± 21.56 mg) compared with the placebo group (29.57 ± 22.38 mg; 95% confidence interval -18.8 to 2.76; p = 0.047). No significant differences were observed in pain evaluated by visual analog scale, except for pain at mobilization 6 h after surgery (ropivacaine versus placebo: 3.90 ± 2.66 versus 5.36 ± 2.55; p = 0.030). No significant differences were observed in the incidence of postoperative nausea/vomiting, pruritus, and time of first ambulation. CONCLUSION: Continuous ropivacaine subfascial wound infusion results in less post-cesarean morphine consumption. EudraCT trail registration number: 2017-004797-33. EudraCT link: https://www.clinicaltrialsregister.eu/ctr-search/trial/2017-004797-33/BE#A.

Cumulative delivery rates in different age groups after artificial insemination with donor sperm.

BACKGROUND: Although the age-effect on in vitro fertilization outcomes has been well documented, data on donor insemination are scarce hampering accurate patient counseling. This cohort study therefore aims at analyzing cumulative delivery rates after donor insemination for various indications. METHODS: A large retrospective analysis was performed on 6630 insemination cycles in 1654 women. Delivery rates were calculated by life-table analysis after a maximum of 12 cycles in five subgroups of age when starting inseminations. Multivariable modeling was used to explore the effects according to age, indication (male infertility, lesbian couple or single-parent request) and ovarian stimulation protocol (none, clomiphene citrate or gonadotrophins). RESULTS: Overall, 928 deliveries were observed, i.e. a delivery rate of 14% per cycle and an expected cumulative delivery of 77% after 12 cycles. Subgroup analysis showed an expected cumulative delivery after 12 cycles of 87% for the group aged 20-29, 77% for ages 30-34, 76% for ages 35-37, 66% for ages 38-39 and 52% for ages 40-45. Drop-out analysis in the latter subgroup showed that only one patient discontinued treatment because of medical reasons. In contrast to age, neither indication nor ovarian stimulation protocol had any significant effect on the delivery rate. CONCLUSIONS: Our study corroborates the impact of age on donor insemination outcome. Nevertheless, even in some older age subgroups, acceptable expected cumulative delivery rates were observed. Despite this, the main reason for discontinuing treatment, however, was the anticipated low success rate. Women, up until 42 years of age, could be encouraged to continue treatment.

Impact of thyroid autoimmunity on cumulative delivery rates in in vitro fertilization/intracytoplasmic sperm injection patients.

OBJECTIVE: To predict the impact of thyroid autoimmunity (TAI) on the probability of delivery after a defined number of treatment cycles, using analysis of cumulative delivery rates in patients with and without TAI. DESIGN: Retrospective cohort study performed at the Center for Reproductive Medicine and Department of Endocrinology, University Hospital of Brussels, approved by the institutional review board of the hospital. SETTING: University hospital. PATIENT(S): All patients who started their first IVF/intracytoplasmic sperm injection cycle at our fertility center between January 1, 2010 and December 31, 2011 were included. MAIN OUTCOME MEASURE(S): Live birth delivery after 25 weeks' gestation was taken as the primary endpoint of our study Cumulative delivery rates were calculated for both groups until six treatment cycles. INTERVENTION(S): All patients (in both groups) received the usual IVF treatment protocols (i.e., antagonist or agonist protocol). RESULT(S): In total 2,406 women who consulted our center were included. We included 333 patients with TAI and 2019 patients without TAI. In the TAI group the crude cumulative delivery rate after six cycles was 47%, whereas the expected cumulative delivery rate was 65%. In our control the crude cumulative delivery rate after six cycles was 47%, whereas the expected cumulative delivery rate was 76%. CONCLUSION(S): Our study did not confirm an influence of TAI status in patients undergoing fertility treatment on cumulative delivery rates after six IVF/intracytoplasmic sperm injection cycles.