Correlations between clinical parameters, blastocyst morphological classification and embryo euploidy.

OBJECTIVE: The aim of the present study was to evaluate clinical and embryo parameters to predict embryo ploidy. METHODS: In this retrospective analysis, we studied 838 biopsied day-5 blastocysts from 219 patients in the period from May 2021 to July 2022. All embryos were morphologically classified before biopsy and were divided into two groups according to genetic test results. Euploid embryos (299) were compared with aneuploid embryos (539) based on maternal age, anti-Mullerian hormone, antral follicle count, and embryo morphology. RESULTS: Maternal age (36.2±3.0) of euploid embryos was lower than maternal age (37.1±2.5) of aneuploid embryos (p<0.0001). AMH levels were higher (3.9±1.2) in the group of euploid embryos than in the group of aneuploid embryos (3.6±1.3, p<0.0001). However, the AFC was not different in the group of euploid embryos (15.3±6.0) compared to the group of aneuploid embryos (14.5±5.9, p=0.07). The presence of aneuploidy was negatively correlated with top embryo quality (embryos 4AA and 4AB). All euploid embryos (299) were top quality versus 331 of 539 (61.49%) aneuploid embryos (p<0.0001). CONCLUSIONS: We found that euploid embryos were associated with lower maternal age, higher AMH levels, and higher quality embryos.

Serum progesterone level in luteal phase improves pregnancy rate in fresh cycles with blastocyst embryo transfer.

OBJECTIVE: To assess the association between serum level of progesterone during stimulation and in the luteal phase with pregnancy rate in a cohort of patients undergoing in vitro fertilization and embryo transfer (IVF-ET) on day 5. METHODS: Retrospective Cohort Study. Patients: 62 infertile women, aged 24-42 years, undergoing ART at our center from May 2019 to May 2021. Progesterone was evaluated during ovarian stimulation on Day 2, Day 6, and Day 8 of stimulation, day of trigger (P4dhCG), and on the day of blastocyst transfer with 5 days of progesterone supplementation (P4d5+). We also calculated the difference of P4d5+ with P4dhCG. (∆P4). Then we divided the patients into two groups based on progesterone serum levels at P4d5+; <10ng/ml (Group A), ≥10ng/ml (Group B). The Student's t-test was performed for continuous variables; Mann-Whitney's Test and Spearman's Test were used where appropriate for categorical variables. p<0.05 was considered statistically significant. RESULTS: There were positive correlations between ßhCG positive with P4d5+ (p<0.001; Rho 0.770) and ∆P4 (p<0.001; Rho 0.703). The pregnancy rate doubled when the serum progesterone level was ≥10ng/ml on the fifth day of progesterone supplementation compared with P4<10ng/ml (44% vs. 21%, respectively). CONCLUSIONS: The pregnancy rate was positively correlated with the serum P4 level on the fifth day of progesterone supplementation and with the difference between the serum progesterone level in the Dd5+ / dhCG. A higher pregnancy rate was observed when serum progesterone level on the fifth day of progesterone supplementation was ≥10ng/ml.

A comparison of the effects of three different estrogen used for endometrium preparation on the outcome of day 5 frozen embryo transfer cycle.

OBJECTIVE: To evaluate the effects of three different estrogen used for endometrium preparation on pregnancy rate, as well as hormone profile on day 5 frozen embryo transfer (FET) cycles. METHODS: Retrospective, observational study. Setting: A tertiary teaching and research private reproductive medicine center. Patients: Ninety patients who were undergoing endometrium preparation for day five frozen embryo transfer cycle (FET). Intervention(s): The women were divided in three groups according to the administration route of estrogen (E2): oral (Primogyna), transdermal patches (Estradot), or transdermal gel (Oestrogel Pump). These administration routines of estrogen are equivalent to 6mg of estradiol daily. All women received 600mg of vaginal progesterone (P) per day (Utrogestan) for luteal phase support. We drew blood samples on starting P day, as well as on beta hCG day for E2 and P measurements. Main Outcome Measure(s): Clinical pregnancy rates (PR). RESULTS: Patient features in the three groups were comparable. There were no significant differences concerning implantation rate, clinical PR, miscarriage rate, multiple-pregnancy rate, or E2 and P levels on starting P day and on beta hCG day. CONCLUSIONS: In FET cycles with oral (Primogyna) or transdermal patches (Estradot), or transdermal gel (Oestrogel Pump), there was no significant difference on pregnancy rates.

Which ovarian reserve marker relates to embryo quality on day 3 and blastocyst; age, AFC, AMH?

OBJECTIVE: The aim of the present prospective study was to evaluate which ovarian reserve marker would be more reliable as the quality of the A + B embryos (day 3 and blastocyst). METHODS: We ran a prospective study with 124 infertile women, aged 24-48 years, from 2017 to 2018. The patients were divided into 3 groups according to age and the subgroups were compared for AMH, AFC, number of A+B embryos. New division of the 3 groups was performed based on the AMH, and the subgroups were compared for age, AFC and number of A+B embryos. Finally, we divided the patients into 3 groups, based on the AFC, and we compared the subgroups for age, AMH and number of A+B embryos. P<0.05 was considered statistically significant. RESULTS: When the 124 patients were divided according to age, we found a significant fall in an A+B embryo quality (day3; blastocyst) after 35 years (p<0.038; p<0.035), and more severely after 37 years (p<0.032; p<0.027). When the 124 patients were divided according to AMH, there was a significant fall in A+B embryo quality (day 3; blastocyst), with AMH<1ng/ml (p<0.023; p<0.021). When the 124 patients were divided according to AFC, there was a significant fall in A+B embryo quality (day 3; blastocyst) with AFC<7 (p<0.025; p<0.023). These markers had significant associations with embryo quality (p<0.005). CONCLUSION: Age, AFC and AMH have significant associations with A +B embryo quality on day 3 and blastocyst.

The impact of dehydroepiandrosterone in poor ovarian responders on assisted reproduction technology treatment.

One in six couples worldwide will experience at least one infertility problem during their reproductive years. Between 5.6% and 35.1% of women will exhibit poor ovarian response. A variety of methods have been applied to improve ovarian response, including dehydroepiandrosterone. In the ovaries, dehydroepiandrosterone promotes follicular development and granulosa cell proliferation by increasing intraovarian androgen concentrations while simultaneously enhancing the level of follicular insulin-like growth factor-1, which promotes folliculogenesis. Dehydroepiandrosterone supplementation may improve in vitro fertilization outcomes and ovarian response in patients with poor ovarian response. However, a few questions still loom over the effectiveness of dehydroepiandrosterone.

A comparison of the effects of three luteal phase support protocols with estrogen on in vitro fertilization-embryo transfer outcomes in patients on a GnRH antagonist protocol.

OBJECTIVE: This study aimed to evaluate the effects of three different luteal phase support protocols with estrogen on the pregnancy rates and luteal phase hormone profiles of patients undergoing in vitro fertilization-embryo transfer (IVF-ET) cycles. A secondary objective was to evaluate which ovarian reserve markers correlated with pregnancy rates. METHODS: This retrospective observational study was carried out at a private tertiary reproductive medicine teaching and research center. The study enrolled 104 patients undergoing intracytoplasmic sperm injection (ICSI) on an antagonist protocol for controlled ovarian hyperstimulation (COH). The women were divided into three groups based on the route of administration of estrogen (E2) for luteal phase support: oral (Primogyna); transdermal patches (Estradott); or transdermal gel (Oestrogel Pump). The administration of estrogen provided the equivalent to 4 mg of estradiol daily. All women received 600mg of vaginal progesterone (P) per day (Utrogestan) for luteal phase support. Blood samples were drawn on the day of hCG administration and on the day of beta hCG testing to measure E2 and P levels. Clinical pregnancy rate (PR) was the main endpoint. RESULTS: The patients included in the three groups were comparable. No significant differences were found in implantation rates, clinical PR, miscarriage rates, multiple-pregnancy rates, E2 or P levels on the day of beta hCG measurement. Concerning ovarian reserve markers, significant correlations between testing positive for clinical pregnancy and AMH (r = 0.66, p<0.0001) and E2 levels on beta hCG measurement day (r = 0.77; p<.0001) were observed. CONCLUSIONS: No significant differences were seen in the pregnancy rates of patients submitted to IVF-ET cycles with GnRH antagonists given oral, transdermal patches, or transdermal gel E2 during the luteal phase. A correlation was found between clinical pregnancy rate and AMH and E2 levels on beta hCG testing day.

Age as A Predictor of Embryo Quality Regardless of The Quantitative Ovarian Response.

BACKGROUND: One determining factor of a successful in vitro fertilization (IVF) cycle is embryo quality. The aim of the present study was to evaluate associations of embryo quality and reserve markers like age, FSH and AMH. MATERIALS AND METHODS: In this prospective study, 120 infertile women, aged 21-44 years, undergoing routine exploration during an unstimulated cycle preceding assisted reproductive technology (ART) at our center were studied prospectively, from February 2011 to December 2014. Descriptive parameters and patient characteristics were reported as mean (SD) or median (range) depending on the distribution. Student's t test was performed for continuous variables, Wilcoxon and Pearson's Test were used for not distributed variables and Fisher's Test was performed for categorical variables. P<0.05 was considered statistically significant. RESULTS: Overall, at the time of investigation, patients had a mean age of 33.03 ± 4.15 years old. On cycle day three, serum anti-Mullerian hormone (AMH) level was 3.50 ± 1.54 ng/mL, serum follicle-stimulating hormone (FSH) level was 6.29 ± 1.53 mUI/ mL, at baseline, women had 16.57 ± 7.0 antral follicles. The mean of collected oocytes was 11.80 ± 5.25, embryo I+II was 2.46 ± 2.11. A greater number of embryos I+II was observed in young patients. By evaluating 120 patients, a significant relationship was observed between age and FSH (r=0.24, P=0.01), age with AMH (r=-0.22, P=0.02), age with collected oocytes (r=-0.23, P=0.03) and age with embryo I+II (r=-0.22, P=0.03). A significant relationship was also observed between antral follicle count (AFC) and AMH (r=0.29, P=0.01), AFC and the number of transferred embryo (r=-0.18, P=0.03), AFC and total dose of the drugs (r=-0.23, P=0.03). Significant relationship of FSH with total dose of drugs (r=0.19, P=0.02) was also observed. In addition, we determined significant relationships between AMH and the number of collected oocytes (r=0.38, P=0.01), AMH and the number of metaphase II oocytes (r= 0.35, P=0.01), AMH and the number of embryo (r=0.19, P=0.04) as well as AMH and total dose of the drugs (r=-0.25, P=0.01). CONCLUSION: Commonly used clinical markers of ovarian reserve are reflection of the ovarian reserve, while the outcome measurements of ART and age are the best predictors of embryo quality.

Early follicle development alters the relationship between antral follicle counts and inhibin B and follicle-stimulating hormone levels on cycle day 3.

OBJECTIVE: To verify whether, during the luteal-follicular transition, increased inhibin B production by abnormally large follicles modifies the expected relationship between the number of follicles and peripheral inhibin B and FSH levels. DESIGN: Prospective study. SETTING: Teaching hospital, France. PATIENT(S): A total of 192 normo-ovulatory women. INTERVENTION(S): Serum inhibin B and FSH levels and numbers and sizes of antral follicles (3-12 mm) were measured on cycle day 3. MAIN OUTCOME MEASURE(S): The strength of hormonal-follicular correlations was assessed in two groups formed according to the presence (large follicle group; n = 73) or absence (small follicle group; n = 119) of one follicle measuring >7 mm. RESULT(S): Serum inhibin B and FSH levels were correlated with antral follicle counts (r = 0.32 and r = -0.44, respectively). These relationships were significantly weaker in the large follicle group (r = 0.24 and r = -0.28, respectively) than in the small follicle group (r = 0.51 and r = -0.55, respectively). CONCLUSION(S): Antral follicle size influences serum inhibin B and FSH levels and alters their expected relationship with the number of antral follicles on day 3. These results contribute to clarifying the controversial role of serum inhibin B and FSH levels in the prediction of ovarian follicular status.

Mock embryo transfer does not affect uterine contractility.

OBJECTIVE: To assess the degree of utero-stimulation induced by transcervical introduction of a catheter as in mock ET. DESIGN: Prospective study. SETTING: Reproductive medicine unit. PATIENT(S): Eighty volunteers undergoing mild ovarian stimulation and IUI. INTERVENTION(S): Just before IUI, women had uterine contraction (UC) frequency assessed with 2-minute sagittal ultrasound scans of the uterus (precatheter measurement). Thereafter, one of three types of catheters, each with a different stiffness level (low, n = 25; medium, n = 30; or high, n = 25) was introduced into the cervix up to the inner cervical os, then immediately withdrawn, and the UC frequency reassessed (postcatheter measurement). MAIN OUTCOME MEASURE(S): Percentage change in UC frequency. RESULT(S): Median (range) UC frequency was not significantly different at pre- and postcatheter measurements: 2.0 (0.0-4.5) UC per minute and 2.2 (0.0-5.5) UC per minute. In addition, the stiffness of the catheter (low, medium, or high) did not influence the UC frequency, with median (range) percentage of change from pre- to postcatheter measurement at 0 (-100% to +75%), 0 (-100% to +100%), and 0 (-40% to +100%), respectively. CONCLUSION(S): The introduction of a catheter up to the inner cervical os, as usually performed in mock ET, does not stimulate uterine contraction frequency, irrespective of catheter stiffness.

[Evidence of follicle responsiveness to FSH by antimüllerian hormone in ovulating women]. / Evidência da resposta folicular ao FSH pelo hormônio antimülleriano em mulheres ovulatórias.

PURPOSE: to test the hypothesis that the anti-müllerian hormone (AMH) serum level reflects the antral follicles' response to the administration of FSH. METHODS: prospective study, including 116 normo-ovulatory infertile patients submitted to controlled ovarian hyperstimulation with GnRH and FSH agonists. The AMH serum level was measured after reaching the pituitary suppression and before the FSH administration (basal day). The number of antral follicles was determined by ultrasonography at the basal day (precocious antral follicles; 2 to 8 mm) and at the day of hCG administration (dhCG; pre-ovulatory follicles; >16 mm). The follicle response to FSH was determined by the percentage of precocious antral follicles which reached pre-ovulatory stage in response to FSH (maturation rate). The correlation of AMH with the patients' age, the total number of precocious antral and pre-ovulatory follicles, collected oocytes, total dose of FHS in the controlled ovarian stimulation and the rate of follicular maturation was studied. For the statistical analysis, it simple regression analysis and the Spearman's test were used, at a 5% significance level. RESULTS: The serum level of AMH was positively correlated with the number of precocious antral follicles at the basal day (r=0.64; p<0.0001) and pre-ovulatory follicles in dhCG (r=0.23; p=0.01). Exceptionally, the serum level of AMH was negatively correlated with the maturation ratio (r=-0.24; p<0.008). CONCLUSIONS: AMH attenuates the follicular development caused by FSH administration.

Evidência da resposta folicular ao FSH pelo hormônio antimülleriano em mulheres ovulatórias / Evidence of follicle responsiveness to FSH by antimüllerian hormone in ovulating women

OBJETIVO: examinar a hipótese de que o nível sérico do hormônio antimülleriano (AMH) reflete a resposta dos folículos antrais à administração do FSH. MÉTODOS: estudo prospectivo, no qual foram incluídas 116 pacientes normo-ovulatórias inférteis submetidas à hiperestimulação ovariana controlada com agonista de GnRH e FSH. Depois de atingir a supressão pituitária e antes da administração de FSH (dia basal), o nível sérico de AMH foi mensurado. O número de folículos antrais foi determinado pela ultrassonografia no dia basal (folículos antrais precoces; 2 a 8 mm) e no dia da administração do hCG (dhCG; folículos pré-ovulatórios; >16 mm). A resposta folicular ao FSH foi determinada pela percentagem de folículos antrais precoces que atingiram os estágios pré-ovulatórios em resposta ao FSH (taxa de maturação). Foram estudadas as correlações do AMH com a idade das pacientes, número total de folículos antrais precoces e pré-ovulatórios, oócitos coletados, dose total de FSH na estimulação ovariana controlada e a taxa de maturação folicular. Para análise estatística, foram usados a regressão simples e o teste de Spearman's, com nível de significância de 5%. RESULTADOS: o nível sérico de AMH foi positivamente correlacionado com o número de folículos antrais precoces no dia basal (r=0,64; p<0,0001) e folículos pré-ovulatórios em dhCG (r=0,23; p=0,01). Excepcionalmente, o nível sérico de AMH foi negativamente correlacionado com a taxa de maturação (r=-0,24; p<0,008). CONCLUSÕES: o AMH atenua o desenvolvimento folicular em resposta à administração do FSH.

PURPOSE: to test the hypothesis that the anti-müllerian hormone (AMH) serum level reflects the antral follicles' response to the administration of FSH. METHODS: prospective study, including 116 normo-ovulatory infertile patients submitted to controlled ovarian hyperstimulation with GnRH and FSH agonists. The AMH serum level was measured after reaching the pituitary suppression and before the FSH administration (basal day). The number of antral follicles was determined by ultrasonography at the basal day (precocious antral follicles; 2 to 8 mm) and at the day of hCG administration (dhCG; pre-ovulatory follicles; >16 mm). The follicle response to FSH was determined by the percentage of precocious antral follicles which reached pre-ovulatory stage in response to FSH (maturation rate). The correlation of AMH with the patients' age, the total number of precocious antral and pre-ovulatory follicles, collected oocytes, total dose of FHS in the controlled ovarian stimulation and the rate of follicular maturation was studied. For the statistical analysis, it simple regression analysis and the Spearman's test were used, at a 5% significance level. RESULTS: The serum level of AMH was positively correlated with the number of precocious antral follicles at the basal day (r=0.64; p<0.0001) and pre-ovulatory follicles in dhCG (r=0.23; p=0.01). Exceptionally, the serum level of AMH was negatively correlated with the maturation ratio (r=-0.24; p<0.008). CONCLUSIONS: AMH attenuates the follicular development caused by FSH administration.

Relação entre os níveis séricos do hormônio anti-Mulleriano, inibina B, estradiol e hormônio folículo estimulante no terceiro dia e o status folicular ovariano / Relationship of serum anti-Müllerian hormone, inhibin B, estradiol and FSH on day 3 with ovarian follicular status

OBJETIVO: examinar a hipótese de que o nível sérico do hormônio anti-Mülleriano (HAM) reflete o status folicular ovariano. MÉTODOS: Desenho: estudo prospectivo. Pacientes: foram incluídas 101 candidatas à FIV-TE submetidas à estimulação ovariana controlada com agonista de GnRH e FSH. Depois de atingir a supressão da hipófise e antes da administração de FSH (dia basal), os níveis séricos de HAM, inibina B e FSH foram avaliados. O número de folículos antrais foi determinado pela ultra-sonografia (dia basal) (folículo antral precoce; 3-10 mm). RESULTADOS: as médias do nível sérico de HAM, inhibina B, E2, P4 e FSH (dia basal) foram 3,4±0,14 ng/mL, 89±4,8 pg/mL, 34±2,7 pg/mL, 0,22±0,23 ng/mL e 6,6±0,1 mUI/mL, respectivamente, e a média do número de folículos antrais precoces foi 17±0,39. O nível sérico do HAM foi negativamente correlacionado com a idade (r= -0,19, p<0,04) e positivamente correlacionado com o número de folículos antrais precoces (r=0,65, p<0,0001), mas isto não se aplicou aos níveis séricos de inibina B, E2 e FSH. CONCLUSÕES: esse dado demonstra a associação do HAM com a quantidade de folículo antral, sendo aquele, portanto, um provável biomarcador do status folicular ovariano.

PURPOSE: to examine the hypothesis that serum anti-Müllerian hormone (AMH) levels reflect the ovarian follicular status. METHODS: Design: prospective study. Patients: we studied 101 IVF-ET candidates undergoing controlled ovarian hyperstimulation with GnRH agonist and FSH. After the achievement of pituitary suppression and before FSH administration (baseline), serum AMH, inhibin B, and FSH levels were measured. The number of antral follicles was determined by ultrasound at baseline (early antral follicles; 3-10 mm). RESULTS: at baseline, median serum levels of AMH, inhibin B, E2, P4 and FSH were 3.42±0.14 ng/mL, 89±4.8 pg/mL, 34±2.7 pg/mL, 0.22±0.23 ng/mL and 6.6±0.1 mIU/mL, respectively, and the mean number of early antral follicles was 17±0.39. Serum levels of AMH were negatively correlated with age (r=-0.19, p<0.04), and positively correlated with number of antral follicles (r=0.65, p<0.0001), but this did not apply to serum levels of either inhibin B, E2 or FSH. CONCLUSION: the data demonstrate an association between AMH and antral follicular counts. Therefore, AMH is probable a biomarker of ovarian follicular status.

Gameta: unidade básica em reproduçäo humana / Gamete: basic unit in human reproduction

Mesmo com tantos avanços na investigação e no tratamento da infertilidade, muitos casais ainda se mantêm infertéis, grande parte devido à idade oocitária das pacientes. A utilização de programas de doação de gametas, além de suprir as necessidades destas pacientes, trouxe vários problemas éticos-legais. Estudos preliminares em animais, utilizando os novos avanços nas técnicas de reprodução assistida, estão apresentando uma nova alternativa de obtenção de gametas com material genético dos próprios pacientes em tratamento. Ficção ou realidade, a ciência avança, e com ela novas perspectivas no tratamento da infertilidade