The impact of severe oligozoospermia on morphokinetic embryo development in low-prognosis patients according to the Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number criteria: an analysis of 10,366 injected oocytes.

OBJECTIVE: To study whether severe male factor infertility (SMF), reflected by oligozoospermia, impacts embryo morphokinetic behavior in low-prognosis women as stratified by the Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number (POSEIDON) criteria. DESIGN: Cohort study. SETTING: Private university-affiliated in vitro fertilization center. PATIENT(S): A total of 10,366 injected oocytes from 2,272 women who underwent intracytoplasmic sperm injection cycles between March 2019 and April 2022. INTERVENTION(S): Patients were divided into 8 groups according to the POSEIDON criteria (1-4) and the presence or absence of SMF. A control group of normoresponder patients was included. Kinetic markers from the point of insemination were recorded in the EmbryoScope incubator. MAIN OUTCOME MEASURE(S): Morphokinetic milestones and intracytoplasmic sperm injection clinical outcomes. RESULT(S): Embryos from patients in the POSEIDON 1 group showed significantly slower timing to pronuclear appearance, timing to pronuclear fading (tPNf), timing to 2 (t2), 3 (t3), 4 (t4), 6 (t6), and 7 (t7) cells than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and non-SMF (nSMF) subgroups in both POSEIDON 1 as well as control groups. Embryos from patients in the POSEIDON 2 group showed significantly slower timing to pronuclear appearance, t4, t6, t7, timing to 8 cells (t8), and timing to morulae than those from the control group. Embryos in the POSEIDON 2 SMF subgroup took longer than those in the POSEIDON 2 nSMF subgroup and those in both control subgroups to achieve tPNf, t2, t3, timing to 5 cells (t5), timing to start blastulation, and timing to blastulation. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 2 as well as control groups. Embryos from patients in the POSEIDON 3 group showed significantly slower t8 and duration of the second cell cycle (t3-t2) than those from the control group. Known Implantation Diagnosis Score ranking was significantly different across the subgroups. Embryos derived from patients in the POSEIDON 4 group showed significantly slower tPNf, t2, t3, t4, t5, t6, t7, t8, timing to complete t4-t3 synchronous divisions, and timing to complete t8-t5 synchronous divisions than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 4 as well as control groups. Irrespective of sperm quality, clinical outcomes significantly improved in the control subgroups compared with those in the POSEIDON 2 and 4 subgroups. CONCLUSION(S): Embryos in the SMF groups presented lower Known Implantation Diagnosis Score ranking than those in the nSMF groups in both POSEIDON 1-4 and control groups, suggesting that cumulative differences result in worse morphokinetic development when the algorithm is used.

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.

Progesterone-primed cycles result in slower embryos without compromising implantation potential and with the advantages of oral administration and potential cost reduction.

OBJECTIVE: To study the impact of the use of progesterone on embryo morphokinetics and on the outcomes of intracytoplasmic sperm injection cycles. DESIGN: Cohort study. SETTING: Private university-affiliated in vitro fertilization center. PATIENT(S): This study included 236 freeze-all intracytoplasmic sperm injection cycles and the resultant 2,768 injected oocytes cultured in a time-lapse imaging incubation system. Patients were matched by age and divided into groups depending on the protocol used to prevent the luteinizing hormone surge: progestin-primed (144 cycles and 1,360 embryos) and gonadotropin hormone-releasing hormone (GnRH) antagonist (144 cycles and 1,408 embryos) groups. INTERVENTION(S): The kinetic recorded markers were time to pronuclear appearance and fading, time to 2-8 cells, time to morulation, time to start of blastulation, and time to blastulation. The durations of cell cycles and time to complete synchronous divisions were calculated. The Known Implantation Data Score ranking was recorded. Morphokinetics and clinical outcomes were compared between the groups. MAIN OUTCOME MEASURE(S): Embryo morphokinetics and clinical outcomes. RESULTS: Slower time to pronuclear appearance, time to 2 cells, time to 7 cells, time to start of blastulation, and time to blastulation were observed in embryos derived from progestin-primed cycles than in those from the GnRH antagonist group. No significant differences were noted in any other morphokinetic milestone. Significantly higher cancellation and implantation rates were observed in the progestin-primed group. However, no significant differences were noted in the pregnancy and miscarriage rates. The expenses for treatment using premature GnRH antagonist and progestins were US$318.18 and US$11.05, respectively. CONCLUSIONS: Exogenous progesterone replaces the GnRH antagonist for the prevention of premature luteinizing hormone surge, in freeze-all cycles, with the advantage of oral administration and potential cost reduction.

The Child's Right to Know Versus the Parents' Right Not to Tell: The Attitudes of Couples Undergoing Fertility Treatments Towards Identity-Release Gamete Donation.

Background: In Brazil, donor anonymity is mandatory; however, the tendency of Brazilians towards the practice is unknown. In this study, an attempt was made to investigate whether couples undergoing assisted reproductive technology (ART) have a different perception of anonymous versus identity-release gamete donation than a target population in Brazil. Methods: This cross-sectional study was performed from September 1, 2020 to December 15, 2020. For that purpose, surveys through online platforms were conducted, including either patients undergoing ART (ART-group, n=400) or subjects interested in the theme (interested-group, n=100) randomized by age at a 1:4 ratio. The survey collected information on the participants' attitudes towards anonymity of gamete donors, and answers were compared between the groups. Results: Most participants stated that the relationship between children and their parents would be affected by the child's knowledge of the origin of its conception. Most participants in the ART-group believed that the gamete donor's identity should not be revealed to the child, while only half of the interested-group stated the same. Most of the participants stated that "the donor's identity should be revealed if the child questions its biological origin". "From birth" was the second most common response, while "when the child turns 18 years old" and "sometime during teenage years" were less common answers. Conclusion: The attitudes of ART patients about anonymity are conservative, with most participants believing that family relationships may be affected if the child is aware of the origin of his/her conception. These patients also believe that the identity of the gamete donor should not be revealed to the child.

Serum anti-Müllerian hormone concentrations are related to embryo development: lessons from time-lapse imaging.

Our objective was to study whether serum anti-Müllerian hormone (AMH) concentrations were associated with embryo morphokinetic events. This retrospective cohort study was performed in a private university-affiliated in vitro fertilization centre between March 2019 and December 2020 and included 902 oocytes cultured in a time-lapse imaging incubator, obtained from 114 intracytoplasmic sperm injection cycles performed. The relationship between AMH concentrations and morphokinetic events was investigated by considering the clustering of data (multiple embryos/patient). Evaluated kinetic markers were time to pronuclei appearance (tPNa) and fading (tPNf), time to two (t2), three (t3), four (t4), five (t5), six (t6), seven (t7), and eight cells (t8), (tSB) and time to the start of blastulation (tSB) and to blastulation (tB). Significant inverse relationships were observed between serum AMH concentrations and tPNf, t3, t4, t5, t6, t7, t8, and tB. The AMH was positively correlated with the KIDScore and implantation rate. Increased serum AMH concentrations correlated with faster embryo development. The clinical implications of this effect on embryo development warrant further investigation.

Time-lapse monitoring: An adjunct tool to select embryos for preimplantation genetic testing.

The goal for the present study was to investigate the effect of aneuploidy on embryo morphokinetics events in a time-lapse imaging (TLI) system incubator. This retrospective cohort study was performed in a private university-affiliated in vitro fertilization center, between 2019 March and December 2020. Kinetic data were analyzed in 935 embryos, derived from 316 patients undergoing intracytoplasmic sperm injection cycle with preimplantation genetic testing (PGT) for aneuploidy, individually cultured in a TLI incubator until Day 5 of development. Timing of morphokinetic variables, the incidences of multinucleation, and Known Implantation Data Score (KIDScore)-Day 5 were compared between euploid (n = 352) and aneuploid embryos (n = 583). Aneuploid embryos showed significantly longer timing to complete specific morphokinetic parameters compared to euploidy embryos. Euploidy embryos also showed a significantly higher KIDScore when compared with the aneuploidy ones. Our evidence suggests that TLI monitoring may be an adjunct approach to select embryos for PGT; however, cautious investigation is still needed.

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.

Previous infection with SARS-CoV-2 impacts embryo morphokinetics but not clinical outcomes in a time-lapse imaging system.

The goal for the present study was to investigate whether previous infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may compromise embryo morphokinetics and implantation. For that, a historical cohort study was performed in a private university-affiliated in vitro fertilization center. The study included 1628 embryos from 88 patients undergoing intracytoplasmic sperm injection (ICSI) cycles. Patients were age-matched in a 1:3 ratio to either a coronavirus disease (COVID) group, including patients with a positive SARS-CoV-2 immunoglobulin test (n = 22 patients, 386 embryos), or a control group, including patients with a negative SARS-CoV-2 immunoglobulin test (n = 66, 1242 embryos). The effect of previous infection with SARS-CoV-2 on morphokinetic events and ICSI outcomes was evaluated. Embryos derived from patients in the COVID group presented longer time to pronuclei appearance and fading, time to form two, three, four and five cells, and time to blastulation. The durations of the third cell cycle and to time to complete synchronous divisions were also significantly increased in the COVID group compared with the control group, whereas known implantation diagnosis score Day 5 ranked significantly lower in the COVID group. No differences were observed between the COVID and control groups on clinical outcomes. In conclusion, patients planning parenthood, who have recovered from COVID-19 infection, must be aware of a possible effect of the infection on embryo development potential.

Ovarian stimulation with luteinizing hormone supplementation: the impact of timing on ovarian response and ICSI outcomes.

OBJECTIVE: To investigate whether the timing of rLH addition to rFSH during controlled ovarian stimulation (COS) impacts ovarian response and the outcomes of intracytoplasmic sperm injection (ICSI) cycles. METHODS: Data of 1278 patients undergoing ICSI between 2015 and 2018, in a private university-affiliated IVF center were analyzed. Patients were divided into groups according to the timing of LH addition to the COS protocol: Group LH-start (n=323), in which LH was administered since day 1 of ovarian stimulation; and Group LH-mid (n=955), in which LH was administered concomitantly with gonadotropin releasing hormone (GnRH) antagonist. Data were also stratified according to female age and response to COS. The outcomes of COS and ICSI were compared between the groups. RESULTS: For the general group and in patients aged ≥ 35 years, higher blastocyst development rates were in Group LH-mid compared to Group LH-start. In patients with poor response to COS (POR), higher fertilization rate, blastocyst development rate and implantation rate were observed in Group LH-mid. CONCLUSIONS: rLH supplementation in POR patients may improve laboratorial and clinical outcomes when started in the mid-follicular phase, in GnRH antagonist ICSI cycles.

High oocyte immaturity rates affect embryo morphokinetics: lessons of time-lapse imaging system.

RESEARCH QUESTION: Does oocyte immaturity rate affect morphokinetic events in a time-lapse imaging (TLI) system? DESIGN: Historical cohort study carried out in a private university-affiliated IVF centre. Injected oocytes (n = 3368) cultured in a TLI incubator, from intracytoplasmic sperm injection (ICSI) cycles (n = 474) carried out between March 2019 and December 2020, were analysed. The effects of immature oocyte rates (the number of germinal-vesicle and metaphase I oocytes by the number of retrieved oocytes in each cycle, on morphokinetic events) were investigated considering clustering of data using mixed models. Evaluated kinetic markers were pronuclei appearance (tPNa), timing to pronuclei fading (tPNf), timing to two (t2), three (t3), four (t4), five (t5), six (t6), seven (t7), and eight cells (t8), timing to morulae (tM) and timing to start of blastulation (tSB) and to blastulation (tB). Durations of the second (t3-t2) and third (t5-t3) cell cycles (cc2 and cc3, respectively) and timing to complete synchronous divisions s1 (t2-tPNf), s2 (t4-t3) and s3 (t8-t5) were also evaluated. RESULTS: Positive relationships were observed between oocyte immaturity rates and slower tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, tSB, tB and cc3. Multinucleation at two- and four-cell stages were positively correlated with oocyte immaturity rate. The KIDScore ranking was negatively correlated with oocyte immaturity rate. No associations were found between oocyte immaturity rate and clinical outcomes. CONCLUSIONS: Increasing oocyte immaturity rate correlates with delayed cell cleavage and blastulation. These findings highlight the importance of TLI for the identification and de-selection of slow-growing embryos for transfer, in cycles with high oocyte immaturity rate.

Improved embryonic development and utilization rates with EmbryoScope: a within-subject comparison versus a benchtop incubator.

The objective of this study was to investigate whether, in consecutive intracytoplasmic sperm injection (ICSI) cycles, embryonic development in an incubator with a time-lapse imaging (TLI) system is better than the previous one obtained in a benchtop incubator (G-185) with similar cultivation characteristics. The study was of a retrospective within-subject design, in which each cycle served as its own control. Data were obtained via the chart review of patients undergoing ICSI in a private university-affiliated in vitro fertilization (IVF) centre who fulfilled the following criteria: second ICSI attempt in which embryos were cultured in a TLI incubator system (TLI group, n = 71), preceded by a first ICSI attempt in which embryos were cultured in a benchtop incubator (Control group, n = 71). Embryonic development up to the fifth day of development, oocyte utilization rate (OUR; transferred embryos plus frozen embryos per total number of retrieved oocytes) and embryo utilization rate (EUR; transferred embryos plus frozen embryos per normally fertilized oocyte) were compared between the groups. There were significant differences in the day 2 non-cleavage rate, day 5 embryo rate, blastocyst development rate, frozen blastocyst rate, OUR, and EUR, in favour of the TLI group. Embryonic development, frozen blastocyst rate, OUR and EUR in the second ICSI cycle were significantly improved when the culture was performed in the EmbryoScope, compared with those rates obtained with culture in a G-185 in the first ICSI cycle of the same patients. The results may also lead to higher cumulative pregnancy outcomes following embryo thawing and transfer.

Paternal ageing impacts blastulation and the outcomes of pregnancy at different levels of maternal age: A clustering analysis of 21,960 oocytes and 3837 ICSI cycles.

The objective of this cross-sectional study was to investigate the impact of paternal age on the outcomes of intracytoplasmic sperm injection (ICSI) cycles at different values of maternal age. A total of 21,960 injected oocytes deriving from 3837 ICSI cycles performed between January 2014 and October 2020, performed in a private university-affiliated in vitro fertilization centre was included. The main effects of maternal and paternal age, as well as the effect of their product (interaction term) on embryo development and pregnancy outcomes were investigated considering the clustering of data. The coefficients for the interaction term were statistically significant for blastocyst development, top-quality blastocyst, implantation, pregnancy, miscarriage, and live-birth rates. For every 1-year increase in paternal age, the odds ratio of live-birth reduces by 1% in females aged 37 years, 1.6% in those aged 38 years, 2.4% in 39-year-old females, 5% in 42-year-old females and so on. An increase in the interaction term by 1 year decreases the pregnancy rate by 0.4% and live-birth rate by 0.8 and increases the miscarriage rate by 1.2%. The slopes of maternal age on blastulation, blastocyst quality, and implantation, pregnancy, miscarriage, and live-birth rate significantly changed (worsened) for every year increase in paternal age.

Dual trigger improves response to ovarian stimulation and ICSI outcomes in patients with a previous r-hCG triggered ICSI cycle.

OBJECTIVE: To evaluate if ovarian response to controlled ovarian stimulation (COS) and intracytoplasmic sperm injection (ICSI) outcomes are improved by the use of dual trigger (gonadotropin-releasing hormone (GnRH) agonists plus recombinant human chorionic gonadotropin (r-hCG)) in patients with previous cycles triggered with r-hCG. METHODS: This case-control study included 88 matched cycles performed in 88 patients, which had the first ICSI cycle triggered with r-hCG (n=44), and the following ICSI cycle with dual trigger (n=44). We compared the cycle outcomes between the groups. In a second case-control within-subject analyses, we compared the ICSI outcomes between patients which had the first ICSI cycle triggered with r-hCG only (n=18), and the following ICSI cycle with dual trigger (n=18) or r-hCG only (n=18). RESULTS: Upon investigating repeated cycles (r-hCG only vs. dual trigger), we found higher oocyte yield and mature oocyte rates, lower immature oocyte rates, higher fertilization rates, and higher blastocyst development rates; and higher rates of cycles with embryos transferred and implantation in the dual trigger cycle. CONCLUSIONS: The dual trigger regimen is a more effective approach than r-hCG trigger in patients with a previous r-hCG triggered ICSI cycle, yielding improved response to COS, and better laboratorial and clinical outcomes.

Maternal lifestyle and nutritional habits are associated with oocyte quality and ICSI clinical outcomes.

RESEARCH QUESTION: Do maternal lifestyle factors influence the incidence of oocyte dimorphisms and outcomes of intracytoplasmic sperm injection (ICSI) cycles? DESIGN: A total of 752 female patients undergoing an ICSI cycle at a private university-affiliated IVF centre from January 2015 to December 2019 were included in this historical cohort study. Before starting ovarian stimulation, participants completed a questionnaire on cigarette smoking habits, consumption of alcoholic beverages, refined sugar, artificial sweeteners, soft drinks, fruits, legumes and vegetables, milk and dairy, and meat, as well as exercise frequency over the past 6 months. Oocyte morphology was evaluated before ICSI. The influence of maternal lifestyle factors on the incidence of oocyte dimorphisms and ICSI outcomes was evaluated by multivariate general linear models and generalized linear models, adjusted for potential confounders. The main outcome measures were the incidence of oocyte dimorphisms per cycle and clinical outcomes. RESULTS: Lifestyle factors and nutritional habits such as cigarette smoking, and the consumption of alcohol, refined sugar and artificial sweeteners, were positively associated with incidence of several oocyte dimorphisms and negatively associated with the response to ovarian stimulation and embryo development. Negative relationships were also observed between these habits and clinical outcomes, apart from miscarriage rate, in which positive relationships were observed. Significant negative dose-dependent relationships between these habits and implantation rates were noted (P < 0.001). Alcoholic beverage consumption also showed inverse dose-dependent relationships with clinical pregnancy and live birth rates. Live birth rate was also negatively associated with cigarette smoking, in a dose-dependent manner. CONCLUSIONS: Poor maternal habits were associated with reduced oocyte quality and ICSI outcomes in this study. Many of these associations were shown to be dose-dependent.

Improving Implantation Rate in 2nd ICSI Cycle through Ovarian Stimulation with FSH and LH in GNRH Antagonist Regimen.

OBJECTIVE: To investigate whether patients with a previous recombinant follicle stimulating hormone (rFSH)-stimulated cycle would have improved outcomes with rFSH + recombinant luteinizing hormone (rLH) stimulation in the following cycle. METHODS: For the present retrospective case-control study, 228 cycles performed in 114 patients undergoing intracytoplasmic sperm injection (ICSI) between 2015 and 2018 in an in vitro fertilization (IVF) center were evaluated. Controlled ovarian stimulation (COS) was achieved with rFSH (Gonal-f, Serono, Geneva, Switzerland) in the first ICSI cycle (rFSH group), and with rFSH and rLH (Pergoveris, Merck Serono S.p.A, Bari, Italy) in the second cycle (rFSH + rLH group). The ICSI outcomes were compared among the groups. RESULTS: Higher estradiol levels, oocyte yield, day-3 high-quality embryos rate and implantation rate, and a lower miscarriage rate were observed in the rFSH + rLH group compared with the rFSH group. In patients < 35 years old, the implantation rate was higher in the rFSH + rLH group compared with the rFSH group. In patients ≥ 35 years old, higher estradiol levels, oocyte yield, day-3 high-quality embryos rate, and implantation rate were observed in the rFSH + rLH group. In patients with ≤ 4 retrieved oocytes, oocyte yield, mature oocytes rate, normal cleavage speed, implantation rate, and miscarriage rate were improved in the rFSH + rLH group. In patients with ≥ 5 retrieved oocytes, higher estradiol levels, oocyte yield, and implantation rate were observed in the rFSH + rLH group. CONCLUSION: Ovarian stimulation with luteinizing hormone (LH) supplementation results in higher implantation rates, independent of maternal age and response to COS when compared with previous cycles stimulated with rFSH only. Improvements were also observed for ICSI outcomes and miscarriage after stratification by age and retrieved oocytes.

OBJETIVO: Investigar se há algum efeito da suplementação com hormônio luteinizante (LH, na sigla em inglês) no regime com antagonista do hormônio liberador de gonadotropina (GnRH, na sigla em inglês) sobre os resultados dos ciclos consecutivos de injeção intracitoplasmática de espermatozoides (ICSI, na sigla em inglês). MéTODOS: Para o presente estudo retrospectivo de caso-controle, foram avaliados 228 ciclos de microinjeção intracitoplasmática de espermatozoides (ICSI, na sigla em inglês) realizados em 114 pacientes entre 2015 e 2018 em um centro privado de fertilização in vitro (FIV) afiliado a uma universidade. O estímulo ovariano controlado (EOC) foi feito com hormônio folículo- estimulante recombinante (rFSH, na sigla em inglês) (Gonal-f, Serono, Genebra, Suíça) no primeiro ciclo de ICSI (grupo rFSH), e com rFSH e rLH (Pergoveris, Merck Serono S.p.A, Bari, Itália) no segundo ciclo (grupo rFSH + rLH). Os desfechos dos ciclos de ICSI foram comparados entre os grupos. RESULTADOS: Níveis mais elevados de estradiol, de recuperação oocitária, taxa de embriões de alta qualidade no 3° dia e taxa de implantação, e menor taxa de aborto foram observados no grupo rFSH + rLH. Em pacientes < 35 anos, a taxa de implantação foi maior no grupo rFSH + rLH em comparação com o grupo rFSH. Em pacientes com ≥ 35 anos, maiores níveis de estradiol, recuperação oocitária, a taxa de embriões de alta qualidade no 3° dia e a taxa de implantação foram observados no grupo rFSH + rLH. Em pacientes com baixa resposta ao EOC (≤ 4 oócitos recuperados), a recuperação oocitária, a taxa de oócitos maduros, a taxa de velocidade normal de clivagem, a taxa de implantação e a taxa de aborto foram melhoradas no grupo rFSH + rLH. Em pacientes com resposta normal ao EOC (≥ 5 oócitos recuperados), níveis mais elevados de estradiol, recuperação oocitária e taxa de implantação foram observados no grupo rFSH + rLH. CONCLUSãO: A estimulação ovariana com suplementação de LH resultou em taxas de implantação mais altas, independentemente da idade materna e da resposta ao EOC, em comparação com os ciclos anteriores estimulados apenas com rFSH. Melhorias também foram observadas nos resultados da ICSI e na taxa de aborto quando as pacientes foram estratificadas por idade e número de oócitos recuperados.

Early and late paternal contribution to cell division of embryos in a time-lapse imaging incubation system.

The objective of this study was to investigate the impact of male age, semen quality and days of ejaculatory abstinence on embryo morphokinetics. A total of 1,220 zygotes obtained from 139 couples in a private in vitro fertilisation centre were analysed. The timing of specific events from the point of insemination, such as timings to pronuclei appearance and fading, to two, three, four, five, six, seven and eight cells and to blastulation were recorded. Multivariate linear regression analysis was used to evaluate the influence of paternal factors on embryo morphokinetic events. Paternal age was positively correlated with delayed cell cleavage and blastulation, and negatively associated with implantation rate, and clinical pregnancy and live-birth chances. The ejaculatory abstinence was inversely correlated with the implantation rate. Inverse relationships were observed between semen parameters (sperm count, progressive sperm motility, total motile sperm count and morphology) and the timing of specific events during embryo development. Sperm morphology was also positively associated with implantation rate and pregnancy and live-birth chances. Increased paternal age and ejaculatory abstinence, and poor semen quality correlate with delayed cell cleavage and blastulation and negatively impact intracytoplasmic sperm injection outcomes.

Current status of the COVID-19 and male reproduction: A review of the literature.

BACKGROUND: Coronavirus disease 2019 (COVID-19), which causes serious respiratory illnesses such as pneumonia and lung failure, was first reported in mid-December 2019 in China and has spread around the world. In addition to causing serious respiratory illnesses such as pneumonia and lung failure, there have been conflicting reports about the presence of SARS-CoV-2 in the semen of patients who were previously diagnosed with COVID-19 and possible implications for the male reproductive tract. OBJECTIVE: The goal for the present study was to review the current status of the literature concerning COVID-19 and male reproduction. MATERIAL AND METHODS: An electronic literature search was done by using PubMed and Google Scholar databases. Relevant papers, concerning SARS-COV-2 and COVID-19 and male reproduction, published between January 2020 and December 2020 were selected, analyzed and eventually included in the present literature review. RESULTS: SARS-CoV-2 may infect any cell type expressing angiotensin-converting enzyme 2 (ACE2), including reproductive cells. Besides the presence of the SARS-CoV-2 receptor, the expression of host proteases, such as transmembrane serine protease 2 (TMPRSS2), is needed to cleave the viral S protein, allowing permanent fusion of the viral and host cell membranes. Here, we aimed to review the current status of the literature concerning COVID-19 and male reproduction. The lack of co-expression of ACE2 and TMPRSS2 in the testis suggests that sperm cells may not be at increased risk of viral entry and spread. However, the presence of orchitis in COVID-19-confirmed patients and compromised sex-related hormonal balance among these patients intrigues reproductive medicine. DISCUSSION: SARS-CoV-2 may use alternate receptors to enter certain cell types, or the expression of ACE2 and TMPRSS2 may not be detected in healthy individuals. CONCLUSION: COVID-19 challenges all medical areas, including reproductive medicine. It is not yet clear what effects, if any, the COVID-19 pandemic will have on male reproduction. Further research is needed to understand the long-term impact of SARS-CoV-2 on male reproductive function.

Cryopreservation of both male and female gametes leads to reduced embryo development and implantation potential.

The objective of this study was to investigate the effect of oocyte and sperm cryopreservation on donated eggs submitted to intracytoplasmic sperm injection (ICSI) cycles. Medical charts of 122 oocyte recipients undergoing 152 oocyte recipient ICSI cycles, from 2017 to 2018, in a private university-affiliated in vitro fertilization (IVF) centre, were reviewed in this historical cohort study. Cycles were divided into four groups according to the gamete status: the FO/FS Group, recipients in which fresh oocytes were injected with fresh sperm (n = 19); the FO/CrS Group, recipients in which fresh oocytes were injected with cryopreserved sperm (n = 14); the CrO/FS Group, recipients in which cryopreserved oocytes were injected with fresh sperm (n = 85); and the CrO/CrS Group, recipients in which cryopreserved oocytes were injected with cryopreserved sperm (n = 34). Generalized mixed models fit by restricted maximum likelihood, followed by Bonferroni post hoc test for the comparison of means amongst groups were used to investigate the effect of cryopreservation on recipient ICSI outcomes. The results were expressed as mean differences with 95% confidence intervals and P-values. The main outcome measure was the implantation rate. Normal day 3 cleavage speed, blastocyst development and implantation rates were significantly lower in the CrO/CrS Group compared with the FO/FS Group. In conclusion, embryo developmental competence and implantation potential were reduced when vitrified oocytes were injected with frozen sperm in an egg-sharing donation programme.

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.

Fresh oocyte cycles yield improved embryo quality compared with frozen oocyte cycles in an egg-sharing donation programme.

The objective of this study was to investigate any effect of cryopreservation of donated eggs on laboratorial and clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles. This retrospective cohort study included 320 oocyte recipients undergoing 307 vitrified and 119 fresh oocyte recipient ICSI cycles, participating in an egg-sharing donation programme, from 2015 to 2018, in a private university-affiliated in vitro fertilization (IVF) centre. A review of donor and recipient ICSI cycles was charted. A general mixed models fit by restricted maximum likelihood, followed by Bonferroni post hoc test was used to compare the means between fresh and warm oocyte donation groups and investigate the effect of cryopreservation on recipient ICSI outcome. The main outcome measure was blastocyst development rates. Fertilization rate, high-quality embryo rates on days 2 and 3, normal cleavage speed rates on days 2 and 3, and blastocyst development rate were significantly higher for the fresh oocyte donation cycles compared with warmed oocyte donation cycles. In the egg-sharing donation programme, fertilization and embryo developmental competence were reduced when vitrified oocytes from infertile couples were used for ICSI compared with fresh oocytes.