F. Belva

Neonatal outcome of 724 children born after ICSI using non-ejaculated sperm

BACKGROUND Safety concerns have been expressed regarding the use of immature non-ejaculated spermatozoa for ICSI. Therefore, adverse health outcomes, birth parameters, major anomaly rates and chromosomal aberrations in children born after ICSI using testicular and epididymal sperm were investigated. METHODS Questionnaire data and results of physical examinations of 530 children born after ICSI with testicular sperm and of 194 children born after ICSI with epididymal sperm were compared with data on 2516 ICSI children born using ejaculated sperm. RESULTS Birth parameters, stillborn rates, prematurity rates and rates of low birthweight and very low birthweight were comparable between the non-ejaculated and the ejaculated sperm groups. The perinatal death rate was higher for twins but not for singletons in the non-ejaculated sperm group in comparison to the control cohort of children born using ejaculated sperm. A non-significant increase in major anomalies was reported in the non-ejaculated sperm group in comparison to the ejaculated sperm group. No more anomalies were observed in pre- and post-natal karyotypes from viable pregnancies established using non-ejaculated sperm versus ejaculated sperm. CONCLUSION Overall neonatal health in terms of birth parameters, major anomalies and chromosomal aberrations in our large cohort of children born by the use of non-ejaculated sperm seems reassuring in comparison to the outcome of children born after the use of ejaculated sperm.

Are ICSI adolescents at risk for increased adiposity

BACKGROUND Puberty is a critical period for the development of cardio-metabolic disturbances, including a more central body fat distribution. It is still unclear if IVF and more specifically ICSI, can permanently and detrimentally affect body fat accumulation in the human offspring. Therefore, adiposity and body fat distribution in 14-year-old adolescents born after ICSI were investigated. METHODS Body composition data, including anthropometry (weight, height and BMI), skinfold thicknesses (peripheral: triceps and biceps skinfolds; central: supra-iliacal and subscapular skinfolds; total: sum of the four skinfolds) and circumferences (waist, mid-upper arm) were compared between 217 ICSI singletons (116 boys, 101 girls) and 223 singletons (115 boys, 108 girls) born after spontaneous conception (SC). ICSI teenagers were part of a previously published ICSI cohort followed since birth; SC controls were recruited from schools in the surroundings. RESULTS Among all boys, no differences in body composition measurements were found between the ICSI and SC group, taking into account confounding variables. In boys with more advanced pubertal stages, a significantly higher sum of peripheral skinfolds was found in the ICSI group compared with the SC group (difference 3.5 mm, 95% confidence interval 0.3-6.6). In girls, peripheral adiposity assessed by skinfolds and mid-upper arm circumference, and central adiposity assessed by skinfolds and waist circumference as well as total adiposity assessed by BMI, the sum of four skinfold thicknesses and skinfold-derived body fat percentage were significantly higher in the ICSI group compared with the SC group, taking into account confounding variables (all P< 0.05). Neither parental nor early life factors could explain the differences. CONCLUSIONS We found that pubertal ICSI girls were more prone to central, peripheral and total adiposity compared with their SC counterparts. ICSI adolescents with advanced pubertal stages showed more peripheral adiposity. Continued monitoring of body fat patterns in adolescents born after fertility treatment is mandatory in order to assess their risk for developing obesity and its related adverse health effects in adulthood.

Blood pressure in ICSI-conceived adolescents

STUDY QUESTION Do young adolescents conceived by ICSI display a higher blood pressure than spontaneously conceived (SC) adolescents? SUMMARY ANSWER In our study, 14-year-old male and female ICSI teenagers were not found to have increased blood pressure at rest. WHAT IS KNOWN AND WHAT THIS PAPER ADDS Only limited data are available regarding the cardiovascular risk of children born after assisted conception and up till now, no data on the cardiovascular health in pubertal children conceived by ICSI have been published. In this study, resting blood pressure and blood pressure response to a psychological stressor were measured in a cohort of 14-year-old teenagers conceived by ICSI and compared the results with those of a group of SC peers. DESIGN In this cross-sectional study, resting blood pressure measurements were available from 217 singleton ICSI children (116 boys, 101 girls) and 223 singleton control children born after spontaneous conception (115 boys, 108 girls). Continuous blood pressure measurements, performed during a psychological stress test, were available for only 67 ICSI and 38 SC children. PARTICIPANTS AND SETTING The study group comprised adolescents conceived by ICSI predominantly because of male factor infertility and they were part of a previously published cohort followed since birth; controls were a cross-sectional sample of peers born to fertile parents and recruited from comparable schools as those attended by the ICSI teenagers. Response rates were 56% (tested/reached) in the ICSI group and 50% (agreed/eligible) in the SC group, but information regarding health could be obtained in 63 and 72% of the ICSI and SC children, respectively. MAIN RESULTS AND THE ROLE OF CHANCE ICSI girls had a comparable resting systolic (109 ± 9 mmHg) and diastolic (64 ± 6 mmHg) blood pressure in comparison with girls in the SC group (111 ± 9 mmHg, P = 0.2 and 66 ± 7 mmHg, P = 0.05), even after adjustment for age and height. After adjustment for current body characteristics, early life and parental background factors, systolic and diastolic blood pressure remained comparable in both groups. In ICSI boys, a slightly lower systolic (113 ± 10 mmHg), but comparable diastolic (64 ± 6 mmHg) resting blood pressure was found in comparison with the SC group (116 ± 9 mmHg; P = 0.04 and 65 ± 5 mmHg; P = 0.1). After adjustment for height and age, systolic and diastolic blood pressure were comparable in both groups (P = 0.7 and P = 0.6). After correction for current body characteristics, early life and parental factors, ICSI and SC boys still had comparable systolic (difference in ICSI versus SC: -1.1 mmHg; 95% CI: -3.8-1.6; P = 0.4) and diastolic (difference in ICSI versus SC: -1.2 mmHg; 95% CI: -3.2-0.7; P = 0.2) blood pressure measurements. In the small subsample of girls and boys with continuous blood pressure readings, the systolic and diastolic blood pressure response to the stress test was not significantly different between the ICSI and SC groups even after taking into account the baseline values. BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION Despite the rather low response rate in the ICSI group and the fact that no information on current health status could be obtained from more than a quarter of the eligible comparison group, the non-participating analysis in the ICSI as well in the SC group did not reveal differences between participating and non-participating children regarding clinical characteristics. The negative results for the sub-analysis on blood pressure response to stress should be interpreted with caution, because these data were available for only a small number of children, and the analysis may be underpowered. This result can only rule out a large effect on blood pressure responsiveness to a psychological stressor. Although our sample size appears to be appropriate, our results need confirmation by others and in larger cohorts when more data become available. GENERALIZABILITY TO OTHER POPULATIONS Our results are the first described ever in ICSI offspring, born to parents suffering from predominantly male factor infertility. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by research grants from Fonds voor Wetenschappelijk Onderzoek Vlaanderen, Onderzoeksraad Vrije Universiteit Brussel and Wetenschappelijk Fonds Willy Gepts. Unconditional grants from MSD Belgium, Merck International, IBSA Institut Biochimique and Ferring International Center are kindly acknowledged.

Pubertal development in ICSI children

BACKGROUND To date, information on the pubertal development of adolescents born after ICSI is scarce, since the very first cohort is only now reaching young adulthood. In this study, pubertal development at the age of 14 was characterized in a longitudinally followed cohort of ICSI-conceived teenagers and compared with that of a spontaneously conceived (SC) control group. METHODS Pubertal development was assessed by Tanner staging (breast, genital and pubic hair development) and age at menarche in 217 singleton ICSI-conceived children (116 boys, 101 girls) and 223 SC peers (115 boys, 108 girls). ICSI teenagers were part of a previously published cohort followed since birth; controls were a cross-sectional sample recruited from schools. Differences in pubertal development between ICSI and SC children were analyzed with logistic regression of current status data. RESULTS Mean age at menarche was similar in ICSI and SC girls (13.1 ± 1.2 versus 13.1 ± 1.4 years; P = 0.8). Breast developmental at the age of 14 years was less advanced in ICSI females compared with SC females, even after adjustment for demographic (age, BMI), genetic (maternal age at menarche), social (maternal educational level) and early life factors (birthweight, gestational age and maternal parity) [odds ratio (OR) 0.5; 95% confidence interval (CI) 0.3-0.8]. After adjustment, genital development in the 14-year-old boys was comparable in the ICSI and SC groups (OR 1.1; 95% CI 0.7-1.8), pubic hair development was comparable in the ICSI and SC groups, for both males (OR 0.9; 95% CI 0.7-1.6) and females (OR 0.7; 95% CI 0.4-1.3). CONCLUSIONS We found that pubertal development, characterized by menarche, genital development in males and pubic hair development in males and females, was comparable in the ICSI and SC groups. Breast developmental was less advanced in ICSI females compared with SC peers, even after adjustment for known potential confounders. In order to confirm that progression through subsequent stages of pubertal development occurs on a timely basis in ICSI teenagers, long-term follow-up studies up to adulthood are required.

Serum inhibin B concentrations in pubertal boys conceived by ICSI: first results

BACKGROUND Currently, no published data exist about the gonadal function of children born after ICSI. To evaluate potential risk of testicular seminal dysfunction in boys born to fathers with compromised spermatogenesis, serum inhibin B (as a marker for spermatogenesis) was assessed. METHODS We recruited 50 pubertal adolescents from the oldest cohort of infants born following ICSI. Cross-sectional serum inhibin B levels of all 50 ICSI adolescents, and longitudinal serum inhibin B (assessed at 8 and 14 years) in 25 boys, are reported. RESULTS A statistically significant increase in inhibin B levels was observed between 8 (mean 69 ng/l, SD ± 35) and 14 years (mean 145 ng/l, SD ± 41; P < 0.001). In three quarters of the ICSI boys an increase in serum inhibin B levels of at least 30% between 8 and 14 years was observed. In all but 4 of the 14-year-old ICSI boys serum inhibin B was normal. Serum inhibin B levels in boys from fathers with severe oligozoospermia did not differ from concentrations in boys from fathers without severe oligozoospermia (154 ± 51 and 142 ± 47 ng/l, respectively; P = 0.4). CONCLUSIONS The majority of ICSI boys have a significant increase in serum inhibin B, attaining normal values for pubertal status at the age of 14 years. ICSI adolescents from fathers with severely compromised spermatogenesis do not have lower inhibin B levels than those with fathers with normal spermatograms. Further follow-up of the spermatogenic potential of ICSI teenagers up to young adulthood is mandatory to confirm a normal reproductive capacity.

Testicular Growth and Tubular Function in Prepubertal Boys Conceived by Intracytoplasmic Sperm Injection

Objective: Little is known about the gonadal function of boys conceived by intracytoplasmic sperm injection (ICSI) from fathers with compromised spermatogenesis. To evaluate the potential risk of tubular dysfunction in these boys, we assessed morphological and functional gonadal parameters and their correlation with paternal sperm characteristics. Study Design: In a group of 88 eight-year-old ICSI boys, we measured testicular and penile size. Serum concentrations of anti-mullerian hormone (AMH) and inhibin B were analyzed in 59 of them. Results: Except for two boys with micropenis, penis length and mean testicular length were normal in all boys. In 7 boys inhibin B concentrations were below the lower limit for age, while all AMH results were within normal limits. Serum Sertoli cell markers correlated significantly with each other (p < 0.005), but were independent of paternal sperm parameters. Conclusion: Our data suggest that penile and testicular growth as well as Sertoli cell function are normal in the majority of prepubertal ICSI boys. Serum AMH and inhibin B levels were found to be independent of sperm quality of the father. Further follow-up of these prepubertal children is needed to examine whether normal Sertoli cell markers will be followed by a normal spermatogenesis in puberty.

Closed oocyte vitrification and storage in an oocyte donation programme: obstetric and neonatal outcome

STUDY QUESTION Does closed oocyte vitrification in an oocyte donation programme have an impact on obstetric and neonatal outcome? SUMMARY ANSWER Obstetric and neonatal outcomes after closed system vitrification of donor oocytes appear to be reassuring. WHAT IS KNOWN ALREADY The use of fresh oocytes has not been proved to be superior to the use of vitrified donor oocytes in terms of survival, embryo development and clinical pregnancies. Those studies used open devices to prove the non-superiority. Very limited information is available on the comparison of open and closed devices, and the results for survival, embryo development and pregnancy outcomes are conflicting. Data on obstetric and neonatal outcome from vitrified oocytes are scarce. Only one large report is available after the use of donor oocytes vitrified with an open device. STUDY DESIGN, SIZE, DURATION Retrospective observational study performed at the Centre for Reproductive Medicine, UZ Brussel, Belgium. All 117 oocyte recipient cycles between March 2010 and August 2014 with the use of a closed vitrification device and leading to a pregnancy beyond 20 weeks were included in this study. PARTICIPANTS/MATERIALS, SETTING, METHODS All recipient warming cycles with a pregnancy beyond 20 weeks from vitrified donor oocytes: results from the fresh embryo transfers. MAIN RESULTS AND THE ROLE OF CHANCE For 117 recipient cycles, a total of 793 oocytes were warmed of which 657 (82.8%) survived and 499 (76.0%) were fertilized. Nineteen single and 98 double embryo transfers led to 95 singleton and 22 twin pregnancies. Hypertensive disorders, haemorrhages and gestational diabetes were reported in 22/112 (19.6%), 30/112 (26.8%) and 13/112 (11.6%) of the pregnancies, respectively. No major adverse neonatal outcomes were observed. Congenital malformations were observed in 11 out of 139 children; for one an elective termination was performed at 25 weeks. LIMITATIONS, REASONS FOR CAUTION Since March 2010, almost all oocytes for donation are vitrified in our centre. Therefore, no recent data are available to control the outcomes of fresh oocyte donations. WIDER IMPLICATIONS OF THE FINDINGS The reassuring results obtained in the current study show that closed system vitrification devices for donor oocytes may be used as an alternative to open devices which have been linked to possible cross-contamination issues. STUDY FUNDING/COMPETING INTERESTS None. TRIAL REGISTRATION NUMBER N/A.

Salivary testosterone concentrations in pubertal ICSI boys compared with spontaneously conceived boys

BACKGROUND To date, no data exist about Leydig cell function of pubertal boys born after ICSI. To evaluate a potential risk of gonadal dysfunction in children born from fathers with compromised fertility, testicular function was assessed by the measurement of salivary testosterone. METHODS Morning salivary testosterone levels at the age of 14 years were compared between 58 ICSI teenagers who are part of the oldest ICSI cohort, and 62 boys born after spontaneous conception (SC). RESULTS Salivary testosterone levels were comparable between ICSI (113 ± 42 pg/ml) and SC (123 ± 56 pg/ml) teenagers at the age of 14 years. In the ICSI group, testosterone levels in boys from fathers with severe oligozoospermia were not different from concentrations in boys from fathers without severe oligozoospermia (115.5 ± 43 and 109 ± 41 pg/ml, respectively). CONCLUSIONS At the age of 14 years, pubertal ICSI boys show testosterone levels comparable to their peers born after SC. ICSI adolescents fathered from men with severely compromised spermatogenesis show testosterone levels comparable to those from fathers with normal spermatogenesis. This notwithstanding, further follow-up of ICSI teenagers into adulthood is mandatory to confirm a normal gonadal function.

Reproductive hormones of ICSI-conceived young adult men: the first results

STUDY QUESTION Are reproductive hormone levels (FSH, LH, inhibin B and testosterone) in male offspring conceived by ICSI because of male infertility comparable with those from peers born after spontaneous conception? SUMMARY ANSWER In this cohort of 54 young men conceived by ICSI because of male-factor infertility, mean and median reproductive hormone levels were found to be comparable with results from spontaneously conceived peers, but ICSI-conceived men were more likely to have low inhibin B (<10th percentile) and high FSH (>90th percentile) levels. WHAT IS KNOWN ALREADY Since the worldwide oldest ICSI offspring have recently reached young adulthood, their reproductive health can now be investigated. This typically involves semen analysis and a hormonal profiling including the measurement of FSH, LH, inhibin B and testosterone. Circulating levels of FSH and inhibin B are generally known as markers of the exocrine function of the testis, i.e. spermatogenesis, while LH and testosterone reflect its endocrine function. We have previously observed a normal pubertal development and comparable levels of inhibin B and testosterone among pubertal ICSI boys when compared to spontaneously conceived peers. However, at present, information on the gonadal function of ICSI offspring in adulthood is still lacking. STUDY DESIGN, SIZE, DURATION This study, conducted between March 2013 and April 2016 at the UZ Brussel, is part of a larger follow-up project focusing on reproductive and metabolic health of young adults between 18 and 22 years and conceived after ICSI because of male infertility. The ICSI men are part of a longitudinally followed cohort while the spontaneously conceived controls were recruited cross-sectionally. PARTICIPANTS/MATERIALS, SETTING, METHODS Results of a single fasting blood sample from 54 young adult ICSI men were compared to that of 57 spontaneously conceived peers. Reproductive hormone analysis involved FSH, LH, testosterone and inhibin B measurement. Furthermore, the association between their reproductive hormones and their sperm parameters was examined. Data were analyzed by multiple linear and logistic regression adjusted for covariates. MAIN RESULTS AND THE ROLE OF CHANCE ICSI men had comparable mean levels of FSH, LH, testosterone and inhibin B in comparison to spontaneously conceived counterparts, even after adjustment for confounders, such as age, BMI and season. Young ICSI-conceived men were more likely to have inhibin B levels below the 10th percentile (<125.2 ng/l; Adjusted Odds Ratio (AOR) 4.0; 95% CI: 0.9–18.4; P = 0.07) compared with spontaneously conceived peers and were more likely to have FSH levels above the 90th percentile (>5.5 IU/L; AOR 3.3; 95% CI: 0.9–11.9; P = 0.06) compared with spontaneously conceived peers, but neither difference reached statistical significance. FSH, LH and inhibin B, but not testosterone, levels were significantly associated with sperm concentration and total sperm count. LIMITATIONS, REASONS FOR CAUTION The main limitation is the small study population. Furthermore, the results of this study should be interpreted according to the background of the participants: all subjects in our study group were conceived by ICSI because of severe male infertility and hence the results cannot be generalized to all ICSI offspring because the indications for performing ICSI have since been widened. WIDER IMPLICATIONS OF THE FINDINGS These first results in a small group of ICSI men show reassuring reproductive hormonal levels. However, larger studies are required to confirm our results. Since inhibin B and FSH are consistently correlated with semen characteristics, we would suggest that the reproductive status of young adults conceived by ICSI is explored with a hormonal assessment given its easier acceptance compared to semen sampling. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by Methusalem grants and by grants from Wetenschappelijk Fonds Willy Gepts, all issued by the Vrije Universiteit Brussel (VUB). A grant from the Belgian Society for Pediatric Endocrinology and Diabetology was received for this project. All co-authors, except M.B. and H.T., declare no conflict of interest. M.B. has received consultancy fees from MSD, Serono Symposia and Merck. The Universitair Ziekenhuis Brussel (UZ Brussel) and the Centre for Medical Genetics have received several educational grants from IBSA, Ferring, Organon, Shering-Plough, Merck for establishing the database for follow-up research and organizing the data collection. The institution of HT receives research grants from the ’Research Fund of Flanders’ (FWO), an unconditional grant from Ferring for research on testicular stem cells and research grants from Ferring, Merck, MSD, Roche, Besins, Goodlife and Cook for several research projects in female infertility. H.T. has received consultancy fees from Finox, Abbott and ObsEva for research projects in female infertility. TRIAL REGISTRATION NUMBER N/A.

Depression, pregnancy-related anxiety and parental-antenatal attachment in couples using preimplantation genetic diagnosis

STUDY QUESTION Do preimplantation genetic diagnosis (PGD) couples experience higher levels of stress during pregnancy and the perinatal period compared with couples who conceive spontaneously (SC) or with ICSI? SUMMARY ANSWER PGD couples did not experience more psychological stress during pregnancy and beyond than ICSI or SC couples. WHAT IS ALREADY KNOWN Previous studies have shown that assisted reproduction technology (ART) couples are more prone to pregnancy-related anxieties than SC couples, but display depressed feelings to an equal or lesser extent. However, only one study has focused on a female PGD sample, which may be a more vulnerable group than other ART groups, due to the potentially complex hereditary background, adverse childhood experiences and losses. In that study, PGD women experienced a reduction in state anxiety, and maternal-antenatal attachment did not differ from normative data. Unfortunately, no data exist on pregnancy-related anxiety, depression and parental-antenatal attachment. Valuable information from both parents (e.g.: couples) is also lacking. STUDY DESIGN, SIZE, DURATION For this longitudinal prospective study questionnaire, data from 185 women and 157 men (157 couples) were collected between February 2012 until April 2014. Data were analysed using multilevel analysis. The couples conceiving after PGD, ICSI or SC were followed from the first trimester of the pregnancy until the third month post-partum. PARTICIPANTS/MATERIALS, SETTING, METHODS A total of 60 PGD, 58 ICSI and 69 SC couples were initially recruited by various departments of Universitair Ziekenhuis Brussel (UZ Brussel). At each trimester (T1: 12-14 weeks, T2: 20-22 weeks, T3: 30-32 weeks) of pregnancy, depression (EPDS), pregnancy-related anxieties (PRAQ) and parental-antenatal attachment (M/PAAS) were recorded. At T4 (3 months post-partum), depression (EPDS) was assessed again. In the first trimester (T1) broad socio-demographic data and at T4 perinatal health data of both mother and child were recorded. Differences between conception groups over time were analysed using multilevel analyses, taking into account covariation between measurements and within couples. Several perinatal covariates as well as social desirability, coping and adult attachment style were controlled for. MAIN RESULTS AND THE ROLE OF CHANCE All three conception groups had similar scores for depression during pregnancy and beyond. Also, pregnancy-related anxiety scales did not differ among the three groups. All groups also followed a similar trajectory in time regarding their scores for anxiety, depression and parental-antenatal attachment. ART groups did not give more socially desirable answers than SC controls. The subsequent moderators: coping and adult attachment style did not add any relevant information. No interaction effects occurred between gender and conception groups. LIMITATIONS, REASONS FOR CAUTION The participants were Caucasian, Dutch-speaking couples, with medium to high socio-economic status, from a single centre. Our data should be replicated by multicultural and multicentre studies. Furthermore, the inclusion of an additional control group of couples who did not opt for PGD but for prenatal diagnosis may point to the most beneficial strategy for the couple. WIDER IMPLICATIONS OF THE FINDINGS PGD parents invest a similar amount of time and emotion in their future children compared with controls. This implies that successful PGD treatment makes an important psychological contribution towards the well-being of couples given their complex hereditary and family backgrounds. STUDY FUNDING/COMPETING INTERESTS This research project was funded by grants from the internal research council of the Vrije Universiteit Brussel (OZR), the Flemish Fonds Wetenschappelijk Onderzoek (FWO) and the Wetenschappelijk Fonds Willy Gepts (WGFG). UZ Brussel and the Centre for Medical Genetics have received several educational grants for organizing the data collection, from IBSA, Ferring, Organon, Shering-Plough, Merck and Merck Belgium. M.B. has received consultancy and speakers fees from Organon, Serono Symposia and Merck.