Association between embryo morphokinetic development and intracytoplasmic sperm injection with epididymal sperm via time-lapse imaging.

The objective of this study was to investigate the impact of sperm source on embryo morphokinetics and the clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles by considering the clustering of data (multiple embryos per patient that share a comparable developmental timing). This matched cohort study was performed at a private university-affiliated in vitro fertilization center. Women who underwent ICSI with epididymal sperm between January 2019 and December 2020 (the percutaneous epididymal sperm aspiration group, n = 32 cycles) were matched with women who underwent ICSI with ejaculated sperm because of idiopathic male factor infertility (the male factor infertility [MFI] group, n = 32 cycles) or female infertility (the control group, n = 32 cycles). Embryos were cultured in a time-lapse imaging incubator, and morphokinetic development was recorded and compared among the groups. Significantly slower divisions were observed in embryos derived from epididymal sperm than in those derived from the MFI and control groups. Embryos derived from epididymal sperm had a significantly lower KIDScore (3.1 ± 0.2) than did those derived from ejaculated spermatozoa from the MFI (5.4 ± 0.1) and control (5.6 ± 0.2, p < 0.001) groups. Epididymal sperm-derived embryos showed a significantly greater occurrence of multinucleation (23.2%) than did those derived from ejaculated sperm from the MFI and control groups (2.8% and 3.7%, p < 0.001, respectively). Epididymal sperm-derived embryos were significantly more likely to undergo direct or reverse cleavage (11.1%) than ejaculated sperm-derived embryos in the control group (4.3%, p = 0.001). In conclusion, delayed cell cleavage and increased incidences of blastomere multinucleation and abnormal cleavage patterns are observed when epididymal-derived sperm are used for ICSI.

Paternal lifestyle factors in relation to semen quality and in vitro reproductive outcomes.

This prospective-cohort study aimed at investigating the influence of paternal lifestyle factors on semen parameters and intracytoplasmic sperm injection (ICSI) outcomes. The influence of paternal lifestyle factors on seminal quality and ICSI outcomes was investigated in male patients undergoing conventional semen analysis. Cigarette smoking negatively influenced semen volume (B: -0.417, slope: 1.570, p = 0.047), sperm count/ml (B: -7.363, slope: 52.298, p = 0.014), total sperm count (B: -4.43, slope: 178.165, p = 0.023), total motile sperm count (B: -1.38, slope: 100.276, p = 0.045) and SDF (B: 0.014, slope: 9.767, p = 0.033). Alcohol consumption negatively influenced sperm count/ml (B: -12.527, slope: 42.255, p = 0.040) and sperm DNA fragmentation (B: 5.833, slope: 9.680, p = 0.002). There were no significant influences of other paternal lifestyle factors. Cigarette smoking negatively influenced the fertilisation rate (B: -1.349, slope: 21.950, p = 0.039) and the blastocyst formation rate (B: -14.244, slope: 28.851, p = 0.025). Alcohol consumption negatively influenced fertilisation rate (B: -3.617, slope: 20.138, p = 0.041) and blastocyst formation rate (B: -34.801, slope: 30.044, p = 0.042). Cigarette smoking and alcohol consumption appear to reduce semen quality, fertilisation and blastocyst formation rates; thus, it would be wise to recommend that male partners reconsider their lifestyle during in vitro reproduction treatment.

The effect of sperm DNA fragmentation on ICSI outcomes depending on oocyte quality.

BACKGROUND: Sperm deoxyribonucleic acid (DNA) fragmentation is commonly encountered in spermatozoa, and the oocyte assumes responsibility for repairing sperm DNA fragmentation during the oocyte-embryo transition. OBJECTIVES: This study aimed to investigate whether the effect of sperm DNA fragmentation on intracytoplasmic sperm injection outcomes depends on the incidence of oocyte dimorphisms. MATERIALS AND METHODS: For the present cohort, 2942 fertilized oocytes from 525 patients submitted to intracytoplasmic sperm injection cycles were assessed. The present study was conducted in a private in vitro fertilization center affiliated to a university from June 2016 to July 2019. Semen samples were divided into the following two groups depending on the sperm DNA fragmentation index: a low fragmentation index group (<30% sperm DNA fragmentation, n = 1468) and a high fragmentation index group (≥30% sperm DNA fragmentation, n = 486). In addition, mature oocytes were examined before sperm injection, and intracytoplasmic and extracytoplasmic defects were recorded. The effect of the sperm DNA fragmentation index on laboratory and clinical intracytoplasmic sperm injection outcomes (depending on the presence of oocyte defects) was evaluated. RESULTS: Significant increases in the rates of fertilization, high-quality embryo, implantation, and pregnancy were noted for cycles with <30% sperm DNA fragmentation than cycles with ≥30% sperm DNA fragmentation (regardless of the presence of oocyte dimorphisms). The presence of dimorphisms significantly impacted laboratory and clinical outcomes. The lowest fertilization and high-quality embryo rates were observed when a high sperm DNA fragmentation index was associated with the presence of dark cytoplasm, vacuoles, resistant membrane, and non-resistant membrane. The lowest implantation and pregnancy rates were observed when a high sperm DNA fragmentation index was associated with the presence of vacuoles, defective perivitelline space, and fragmented polar body. The effect of sperm DNA fragmentation on miscarriage rates was significantly influenced by the presence of centrally located cytoplasmic granulation, a defective perivitelline space and non-resistant membrane. CONCLUSION: A high sperm DNA fragmentation index increases the likelihood of miscarriage in intracytoplasmic sperm injection cycles, an effect that may potentially be magnified by the presence of oocyte dysmorphisms.

Morphokinetic parameter comparison between embryos from couples with high or low sperm DNA fragmentation index.

OBJECTIVE: To study whether time-lapse imaging can identify morphokinetic events impacted by a high sperm DNA fragmentation index (DFI). DESIGN: Historical cohort study. SETTING: Private university-affiliated in vitro fertilization center. PATIENT(S): A total of 978 zygotes cultured until day 5 in a time-lapse imaging incubator between March 2019 and August 2020, derived from 118 patients undergoing intracytoplasmic sperm injection as a result of idiopathic male factor infertility. INTERVENTION(S): Kinetic markers from the point of insemination were recorded. Generalized linear mixed models adjusted for potential confounders followed by the Bonferroni post hoc test were used to compare the timing of specific events in patients with a low (<30%) or high (≥30%) sperm DFI. The recorded kinetic markers were the following: timing to pronuclei appearance and fading; timing to 2, 3, 4, 5, 6, 7, and 8 cells; and timing to start blastulation and blastulation. MAIN OUTCOME MEASURE(S): Timing to blastulation. RESULT(S): Embryos derived from sperm samples with ≥30% DFI showed significantly slower divisions compared with those with <30% DFI (mean differences of 0.7 hours in timing to pronuclei appearance, 1.2 hours in timing to pronuclei fading, 1.5 hours in timing to 2 cells, 2.5 hours in timing to 3 cells, 1.8 hours in timing to 4 cells, 3.3 hours in timing to 5 cells, 3.1 hours in timing to 6 cells, 3.2 hours in timing to 7 cells, 2.7 hours in timing to 8 cells, 8.4 hours in timing to start blastulation, and 3.8 hours in timing to blastulation). The incidences of reverse or direct cleavages (9.3% vs. 4.4%; odds ratio [OR], 2.24; 95% confidence interval [CI], 1.32-3.77) and multinucleation at 2-cell (18.9% vs. 12.0%; OR, 1.70; 95% CI, 1.12-2.58) and 4-cell (14.2% vs. 6.4%; OR, 2.42; 95% CI, 1.57-3.74) stages were significantly higher in embryos deriving from ≥30% DFI than from <30% DFI. The KIDScore ranked significantly different between embryos derived from samples with <30% and ≥30% DFI. Continuous DFI was positively correlated with all timings of specific events and with the incidences of abnormal cleavage patterns (OR, 1.042; 95% CI, 1.025-1.059) and multinucleation at 2-cell stage (OR, 1.053; 95% CI, 1.030-1.076) and inversely correlated with the KIDScore rank (B, -0.218; 95% CI, -0.044 to -0.007). No significant differences were observed in clinical outcomes between the groups. CONCLUSION(S): Embryo morphokinetic parameters are negatively impacted by high sperm DFI, resulting in delayed cell cleavage and blastulation.

Oocyte ability to repair sperm DNA fragmentation: the impact of maternal age on intracytoplasmic sperm injection outcomes.

OBJECTIVE: To study the impact of sperm DNA fragmentation (SDF) on clinical outcomes of assisted reproductive technology in women with different age ranges. DESIGN: Historical cohort study. SETTING: Private university-affiliated in vitro fertilization center. PATIENT(S): Five hundred forty couples undergoing intracytoplasmic sperm injection cycles. INTERVENTION(S): Cycles were split into three groups according to maternal age: ≤36 years old (n = 285), 37-40 years old (n = 147), and >40 years old (n = 108). Semen samples were evaluated for SDF using the Sperm Chromatin Dispersion test and, for each age group, the cycles were subdivided according to SDF index: low fragmentation index (<30% SDF) and high fragmentation index (≥30% SDF). MAIN OUTCOME MEASURE(S): Implantation, pregnancy, and miscarriage rates. RESULT(S): For younger patients (≤36 years old) and those between 37 and 40 years of age, no significant differences were noted in laboratory and clinical outcomes for cycles with <30% SDF or ≥30% SDF. When maternal age was >40 years of age, significantly lower high-quality day-3 embryos (54.4% vs. 33.1% and blastocyst development rates (49.6% vs. 30.2%), lower pregnancy (20.0% vs. 7.7%) and implantation rates (19.7% vs. 11.9%), and increased miscarriage rate (12.5% vs. 100.0%) were observed for cycles with ≥30% SDF compared with <30% SDF, respectively. CONCLUSION(S): Older oocytes, when injected with sperm derived from samples with high SDF index, develop into embryos of poor quality that lead consequently to lower implantation and pregnancy rates and higher miscarriage rates, in intracytoplasmic sperm injection cycles from women with advanced maternal age.

Sperm DNA fragmentation is correlated with poor embryo development, lower implantation rate, and higher miscarriage rate in reproductive cycles of non-male factor infertility.

OBJECTIVE: To study the implications of sperm DNA fragmentation (SDF) in intracytoplasmic sperm injection cycles for non-male factor infertility. DESIGN: Prospective cohort study. SETTING: Private university-affiliated IVF center. PATIENT(S): Data from 475 cycles performed from June 2016 to June 2017. INTERVENTION(S): Cycles were divided according to SDF rate into two groups: <30% SDF (n = 433) and ≥30% SDF (n = 42). Laboratory and clinical outcomes were compared between groups by generalized linear models adjusted for potential confounders. MAIN OUTCOME MEASURE(S): Embryo quality and miscarriage rates. RESULT(S): Fertilization rate was similar between groups (≥30% SDF, 85.28% ± 1.06% vs. <30% SDF, 90.68% ± 3.61%). Significantly lower rates of normal cleavage speed (≥30% SDF, 61.12% ± 4.21% vs. <30% SDF, 72.53% ± 1.24%), high-quality embryos at day 3 (≥30% SDF, 23.07% ± 5.56% vs. <30% SDF, 36.41% ± 1.53%), blastocyst formation (≥30% SDF, 39.09% ± 2.73% vs. <30% SDF, 58.83% ± 7.59%), blastocyst quality (≥30% SDF, 11.97% ± 1.22% vs. <30% SDF, 30.09% ± 2.39%), and implantation (33.24% ± 1.66% vs. <30% SDF, 46.40% ± 4.61%) were observed in cycles with higher SDF, despite similar pregnancy rates (≥30% SDF, 30.40% vs. <30% SDF, 32.40%). A 2.5-fold miscarriage rate was observed in cycles with an SDF above the established cutoff (≥30% SDF, 42.8% vs. <30% SDF, 16.8%). CONCLUSION(S): Higher SDF is correlated with poor embryo development, lower implantation rate, and higher miscarriage rate in non-male factor infertility intracytoplasmic sperm injection cycles. Since defects in sperm may be hidden, the SDF test can bring additional information to the sperm quality evaluation of men with unknown infertility history.

Zika Virus Outbreak - Should assisted reproduction patients avoid pregnancy?

OBJECTIVE: To discuss the requirement from the National Health Surveillance Agency (ANVISA), for assisted reproduction treatment patients to undergo laboratory tests for ZIKV detection, and if the public health authorities and government leaders' recommendations to women simply avoid pregnancy is prudent. METHODS: This study was performed in a university-affiliated in vitro fertilization center in Brazil. We present a critical discussion on the risk of microcephaly due to ZIKV infection and the prevalence of other harmful pathogens to vulnerable pregnant women and infants. We assessed, 954 patients undergoing intracytoplasmic sperm injection cycles (ICSI), between April and November of 2016, concerning the results of ZIKV test, according to different regions in Brazil. RESULTS: Patients undergoing ICSI cycles were split into groups, according to their region of origin: 28 (3.0%) were from the North, 27 (2.8%) were from the Northeast, 40 (4.2%) were from the Midwest, 830 (87.2%) were from the Southeast, and 29 (3.0%) were from the South. Concerning the diagnosis, 112 samples had a positive or inconclusive result for ZIKV, by chromatography immunoassay. These samples were re-analyzed by ELISA and no result was positive. All positive results were from the Southeast region and none from the Northeast or Midwest regions, which are considered endemic regions. CONCLUSION: ZIKV test before the onset of assisted reproduction treatments does not rule out the risk of the infection during pregnancy. In addition, although ZIKV infection risk is extremely high, the microcephaly risk due to ZIKV is not higher than the risk of miscarriage and birth defects due to other recognized pathogens.