Is perivitelline space morphology of the oocyte associated with pregnancy outcome in intracytoplasmic sperm injection cycles?

OBJECTIVE(S): To investigate the effect of perivitelline space (PVS) abnormalities on the outcomes of intracytoplasmic sperm injection (ICSI) cycles in which the entire cohort was affected. STUDY DESIGN: Data from 9752 oocytes obtained from 1151 ICSI cycles performed from June/2010 to August/2016 in a private university-affiliated IVF centre. Cycles were divided into four groups according to the presence or absence of PVS abnormalities: PVS-L group (cycles with the entire oocyte cohort affected by large PVS, n = 265), PVS-G group (cycles with the entire oocyte cohort affected by PVS granularity, n = 280), PVS-L + PVS-G group (cycles with the entire oocyte cohort affected by PVS-L and PVS-G, n = 204), and control group (cycles with the entire oocyte cohort free of PVS abnormalities, n = 402). The effect of PVS abnormalities on ICSI outcomes was assessed by GLM adjusted for potential confounders. RESULTS: Groups with PVS abnormalities presented substantially higher FSH/follicle (p < 0.001) and FSH/oocyte (p < 0.001) ratios, and lower numbers of follicles (p < 0.001), oocytes (p < 0.001) and embryos (p = 0.002) compared to the control group. PVS-L + PVS-G implantation (p = 0.044) and pregnancy (p = 0.004) rates were significantly lower than in cycles with isolated PVS abnormalities and controls. CONCLUSION(S): Cycles in which the entire oocyte cohort is affected by both large PVS and PVS granularity have compromised implantation and pregnancy rates.

Sperm morphological normality under high magnification predicts laboratory and clinical outcomes in couples undergoing ICSI.

The objective of this study was to evaluate whether 'motile sperm organelle morphology examination' (MSOME) is correlated with the outcome of ICSI. A total of 14400 spermatozoa from 72 couples undergoing ICSI were analysed by MSOME (x6600) and graded into four groups: grade I, normal form and no vacuoles; grade II, normal form and lesser than or equal to 2 small vacuoles; grade III, normal form greater than 2 small vacuoles or at least one large vacuole and grade IV, large vacuole and abnormal head shapes or other abnormalities. The correlations between the proportion of morphologically normal spermatozoa (grade I + II) and ICSI outcomes were assessed. The proportion of grade I+ II spermatozoa was lower in patients with oligoasthenoteratozoospermia (OAT) compared to patients with other types of semen alterations (10.6% vs. 17.0%, p = 0.001). The proportion of grade I+ II spermatozoa was positively correlated with blastocyst formation (S = 8.31, R(2):13.5%, p = 0.014) and implantation rates (S = 8.32, R(2): 7.9%, p = 0.030). The proportion of grade I + II spermatozoa was higher in patients with ongoing pregnancy in comparison with those who had a miscarriage (23.2% vs. 10.8%, p = 0.007). Sperm morphological normality was lower in oligoasthenoteratozoospermia patients but correlated with blastocyst formation, implantation and miscarriage rates in couples undergoing ICSI. MSOME may be valuable in predicting ICSI outcomes.

The prevalence of sperm with large nuclear vacuoles is a prognostic tool in the prediction of ICSI success.

PURPOSE: To investigate if there is a correlation between the prevalence of sperm with large nuclear vacuoles (LNV) and intracytoplasmic sperm injection (ICSI) outcomes. METHODS: Two hundred male patients undergoing ICSI had their sperm morphology evaluated through motile sperm organelle morphology examination (MSOME) and the percentage of LNV sperm was recorded and correlated to the ICSI outcomes. RESULTS: The percentage of sperm with LNV negatively influenced the blastocyst formation (S: 16.9, R(2): 20.5%, p = 0.004) and implantation (S: 34.7, R(2): 26.2%, p = 0.001). There were significant differences in the percentage of sperm with LNV between patients in which pregnancy was achieved or not (22.2% vs. 28.4%, p < 0.001) and in patients with ongoing pregnancy or not (22.4% vs. 28.5%, p < 0.001). The incidence of sperm with LNV was determinant to the decreased odds of pregnancy (OR: 0.74, p < 0.001) and increased odds of miscarriage (OR: 1.46, p < 0.001). The area under the curve (AUC) was sufficient to distinguish between couples which did achieve pregnancy or not (AUC: 0.922, p < 0.001). CONCLUSIONS: The MSOME is a prognostic tool in the prediction of ICSI success and could be used to select patients that should have their sperm selected by MSOME for ICSI.

Do weekend oocyte retrievals have an impact on the intracytoplasmic sperm injection cycles outcome?

INTRODUCTION: Despite the fact that ovarian stimulation is controlled, it is not always predictable. Because the day of human chorionic gonadotrophin (hCG) injection depends mainly on the patient's ovarian response to gonadotrophins, the day of oocyte retrieval cannot be determined in advance. As a result, oocyte retrievals are often scheduled to occur on weekends, a fact that entails at least one extra working day for the staff, and could lead to physical and psychological stress, especially in embryologists. The aim of this study was to evaluate whether intracytoplasmic sperm injection (ICSI) outcomes are influenced by the day of oocyte retrieval. MATERIAL AND METHODS: A total of 327 ICSI cycles, whose retrievals were performed on Wednesdays and Sundays, were analysed in this retrospective study. Cycles were subdivided into two groups according to the day of oocyte retrieval: group W (n = 196), cycles in which oocyte retrieval was performed on Wednesday; and group S (n = 131), cycles in which oocyte retrieval was performed on Sunday. Groups were compared regarding fertilization, implantation, pregnancy and take-home baby rates. RESULTS: No significant differences were observed between groups A and B regarding fertilization rate (68.9% and 72.5%; p = 0.1589), implantation rate (21.8% and 24.3%; p = 0.5714), pregnancy rate (29.9% and 31.6%; p = 0.7129) and take-home baby rate (23.6% and 28.1%; p = 0.4351). CONCLUSIONS: A well-trained embryologist's group adhering to staff scheduling allows large programmes to ensure a similar outcome independent of the workload or workday on which embryologists perform the manipulation of gametes.

Are poor responders patients at higher risk for producing aneuploid embryos in vitro?

PURPOSE: To test the hypothesis that aged women with poor ovarian response express an increase on embryo chromosomal alterations when compared to aged women who presented normal response. METHODS: Couples undergoing intracytoplasmic sperm injection cycles with preimplantation genetic screening, were subdivided into two groups: Poor Responder group (n = 34), patients who produced ≤4 oocytes; and Normoresponder group (n = 50), patients who produced ≥5 oocytes. Groups were compared regarding cycles' outcomes and aneuploidy frequency. RESULTS: There were no significant differences between and groups regarding the fertilization rate (p = 0.6861), clinical pregnancy (p = 0.9208), implantation (p = 0.6863), miscarriage (p = 0.6788) and the percentage of aneuploid embryos (p = 0.270). Embryo transfer rate was significantly lower on poor responder group (p = 0.0128) and logistic regression confirmed the influence of poor response on the chance of embryo transfer (p = 0.016). CONCLUSIONS: Aged females responding poorly to gonadotrophins are not at a higher risk for producing aneuploid embryos in vitro.

The effect of GnRH analogues for pituitary suppression on ovarian response in repeated ovarian stimulation cycles.

INTRODUCTION: Ovarian stimulation is employed in assisted reproduction techniques in order to obtain as many oocytes as possible. The early rise in oestradiol levels may lead to the premature end of the respective cycle. In order to avoid such an effect, pituitary suppression has been employed. The aim of this study was to evaluate whether maintenance or replacement of the type of GnRH analogue (i.e., agonist or antagonist) employed for pituitary suppression in the consecutive intracytoplasmic sperm injection (ICSI) cycle would negatively influence oocyte quality and ICSI outcome. MATERIAL AND METHODS: A retrospective observational study was conducted including 181 women with primary infertility. Patients were divided into four different groups according to the GnRH analogue used for pituitary suppression in the first and consecutive cycle. RESULTS: When a GnRH agonist was employed for pituitary suppression in the first cycle, the consecutive cycle showed comparable outcomes when performed with either a GnRH agonist or a GnRH antagonist. When the first cycle was performed with a GnRH antagonist, the use of the GnRH agonist in the successive cycle led to an increased number of oocytes retrieved (7.5% vs. 10.3%, p = 0.032) and the production of a higher number of embryos (4.5% vs. 6.3%, p = 0.036). CONCLUSIONS: When the first cycle is carried out with a GnRH antagonist, the use of a GnRH agonist in the successive cycle would lead to increased numbers of oocytes collected and embryos produced.

Decreased fertility in poor responder women is not related to oocyte morphological status.

INTRODUCTION: In women showing impaired fertility, a decreased response to ovarian stimulation is a major problem, limiting the number of oocytes to be used for assisted reproduction techniques (ART). Despite the several definitions of poor response, it is still a matter of debate whether young poor responder patients also show a decrease in oocyte quality. The objective in this study was to investigate whether poor ovarian response to the superstimulation protocol is accompanied by impaired oocyte quality. MATERIAL AND METHODS: This study included 313 patients younger than 35 years old, undergoing intracytoplasmic sperm injection. Patients with four or fewer MII oocytes (poor-responder group, PR, n = 57) were age-matched with normoresponder patients (NR, n = 256). RESULTS: A higher rate of oocyte retrieval and a trend towards an increase in MII oocyte rate were observed in the NR group when compared to the PR group (71.6 ±1.1% and 74.1 ±1.0% vs. 56.3 ±2.9% and 66.5 ±3.7%; p< 0.0001 and p = 0.056, respectively). A trend toward increased implantation rates was observed in the NR group when compared to the PR group (44 and 24.5 ±2.0% vs. 28.8 and 16.4 ±3.9%; p= 0.0305 and p= 0.0651, respectively). CONCLUSIONS: Low response to ovarian stimulation is apparently not related to impaired oocyte quality. However, embryos produced from poor responder oocytes show impaired capacity to implant and to carry a pregnancy to term.

Blastomere multinucleation: Contributing factors and effects on embryo development and clinical outcome.

INTRODUCTION: The aim of the study was to discover which intracytoplasmic sperm injection (ICSI) cycle parameters could influence the presence of multinucleated blastomeres (MNBs) and how ICSI outcomes are influenced by this event. MATERIAL AND METHODS: Embryos derived from normally fertilised oocytes were divided into two groups: embryos that had only mononucleated blastomeres (NBs group n = 2818) and embryos that had at least one multinucleated blastomere (MNB group, n = 404). The effects of ICSI cycle factors on multinucleation were investigated and embryo development was compared between the groups. The cycles were also split into those in which only NB embryos were present (NB cycles, n = 298) and cycles in which MNB embryos were present (MNB cycles, n = 203). ICSI outcomes were compared between the groups. RESULTS: A higher incidence of MNB embryos arose in pituitary blockage with gonadotropin-releasing hormone agonists, male factor infertility and in cycles with higher number of retrieved oocytes. Embryos that had only one affected blastomere showed greater development than embryos with more than one affected blastomere. Finally, the implantation rate decreased when MNB embryos were transferred. CONCLUSION: Multinucleation events may be affected by aspects of the ICSI cycle and compromise embryo quality and implantation rate.

Intracytoplasmic sperm injection outcome versus intracytoplasmic morphologically selected sperm injection outcome: a meta-analysis.

The development of a modified intracytoplasmic sperm injection (ICSI), called intracytoplasmic morphologically selected sperm injection (IMSI), demonstrated that a profound morphological investigation of the spermatozoon, under the magnification of 6600 x, enables outcome improvement. The aim of this study was to compare ICSI outcome with IMSI outcome. The meta-analysis results demonstrated no significant difference in fertilization rate between ICSI and IMSI groups. However, a significantly improved implantation (odds ratio (OR) 2.72; 95% confidence interval (CI) 1.50-4.95) and pregnancy rate (OR 3.12; 95% CI 1.55-6.26) was observed in IMSI cycles. Moreover, the results showed a significantly decreased miscarriage rate (OR 0.42; 95% CI 0.23-0.78) in IMSI cycles as compared with ICSI cycles. This is the first meta-analysis of published data to evaluate the potential benefits of IMSI. The pooled data of IMSI cycles demonstrate a statistically significant improvement in implantation and pregnancy rates and a statistically significant reduction in miscarriage rates. However, more randomized controlled trials are needed to confirm these results.

Development of secondary palate requires strict regulation of ECM remodeling: sequential distribution of RECK, MMP-2, MMP-3, and MMP-9.

We have evaluated RECK (reversion-inducing-cysteine-rich protein with Kazal motifs), MMP-2 (matrix metalloproteinase-2), MMP-3, and MMP-9 involvement during palate development in mice by using various techniques. Immunohistochemical features revealed the distribution of RECK, MMP-2, and MMP-3 in the mesenchymal tissue and in the midline epithelial seam at embryonic day 13 (E13), MMPs-2, -3, and -9 being particularly expressed at E14 and E14.5. In contrast, RECK was weakly immunostained at these times. Involvement of MMPs was validated by measuring not only their protein expression, but also their activity (zymograms). In situ hybridization signal (ISH) for RECK transcript was distributed in mesenchymal and epithelial regions within palatal shelves at all periods evaluated. Importantly, the results from ISH analysis were in accord with those obtained by real-time polymerase chain reaction. The expression of RECK was found to be temporally regulated, which suggested possible roles in palatal ontogeny. Taken together, our results clearly show that remodeling of the extracellular matrix is finely modulated during secondary palate development and occurs in a sequential manner.

Zona pellucida birefringence in in vivo and in vitro matured oocytes.

OBJECTIVE: To evaluate zona pellucida birefringence (ZPB) in immature and mature oocytes collected after controlled ovarian stimulation and to assess the influence of ZPB on oocyte development. DESIGN: Prospective study. SETTING(S): Private assisted reproduction centre. PATIENT(S): Thirty patients undergoing intracytoplasmic sperm injection. INTERVENTION(S): The ZPB of mature and immature oocytes was evaluated using a polarization imaging software module, and the oocytes were classified as high birefringence (HB) or low birefringence. MAIN OUTCOME MEASURE(S): The ZPB of in vivo and in vitro matured oocytes and its influence on spontaneous nuclear maturation in vitro, fertilization, and embryo quality. RESULT(S): The percentage of HB oocytes was higher in immature than in mature oocytes (40.1 vs. 23.6%). Among immature oocytes, an increased percentage of HB in prophase-I stage oocytes compared to metaphase I stage oocytes was also observed (50.7 vs. 25.0%). However, the percentage of HB oocytes did not change when comparing oocytes before and after in vitro maturation for both prophase I and metaphase I oocytes. No influence of ZPB was observed on the spontaneous in vitro maturation potential. Exclusively for metaphase II retrieved oocytes, a positive influence of ZPB on fertilization (odds ratio [OR], 1.78; 95% confidence interval [CI], 1.27-2.49) and embryo quality (OR, 2.28; 95% CI, 1.04-4.99) was noted. CONCLUSION(S): ZPB may be a useful tool to predict embryo quality for metaphase-II oocytes. Moreover, the completion of nuclear changes in the production of metaphase-II oocytes in vitro may not reflect their molecular maturity.

Zona pellucida birefringence score and meiotic spindle visualization in relation to embryo development and ICSI outcomes.

The meiotic spindle and the zona pellucida exhibit molecular order when imaged with polarized optics. This study aimed to investigate possible factors contributing to the zona pellucida birefringence score and meiotic spindle visualization, and to evaluate whether these structures may predict intracytoplasmic sperm injection outcomes. Oocytes were divided into groups according to zona pellucida birefringence and meiotic spindle visualization. In addition, the cycles were split into three groups based on the zona birefringence of transferred embryos. A positive correlation was observed between zona birefringence and meiotic spindle visualization. In addition, when the meiotic spindle was observed, the fertilization rate among oocytes with high or low zona pellucida birefringence was similar. Implantation and pregnancy rates were significantly higher when embryos derived from high birefringence oocytes were exclusively transferred (P = 0.041 and P = 0.004 respectively). Furthermore, the miscarriage rate was higher when embryos derived from low birefringence oocytes were exclusively transferred. On the other hand, the total dose of FSH negatively affected meiotic spindle visualization. Results show that selection of embryos based on zona pellucida and meiotic spindle imaging can significantly improve implantation and pregnancy rates. Moreover, the dose of FSH used for ovarian stimulation may affect the organization of the oocyte meiotic spindle.

A comparison of post-thaw results between embryos arising from intracytoplasmic sperm injection using surgically retrieved or ejaculated spermatozoa.

OBJECTIVE: To study the effect of freeze-thaw on embryos derived from intracytoplasmic sperm injection (ICSI) using surgically retrieved and ejaculated spermatozoa. DESIGN: Retrospective study. SETTING: Private IVF center. PATIENT(S): Three hundred eighty-three patients undergoing frozen-thawed ET cycles. INTERVENTION(S): Testicular sperm aspiration (TESA) or percutaneous epididymal sperm aspiration (PESA) were the sperm surgical retrieval methods used for ICSI. Embryos resulting from ICSI using surgically retrieved and ejaculated spermatozoa were frozen, thawed, and transferred. MAIN OUTCOME MEASURE(S): Post-thaw survival, implantation, and pregnancy rates. RESULT(S): No differences were found between the ejaculated sperm and TESA/PESA groups in terms of post-thaw survival rate (68.4% vs. 66.1%, respectively), pregnancy rate (20.1% vs. 16.1%), and implantation rate (10.6% vs. 12.7%). Similar results were found for those variables when comparing TESA and PESA groups. CONCLUSION(S): Cleavage embryos arising from ICSI cycles using testicular and epididymal spermatozoa can be frozen with survival, pregnancy, and implantation rates comparable to those obtained with ejaculated spermatozoa.