Influence of a hyaluronan-binding system for sperm selection in intracytoplasmic sperm injection cycles on embryo morphokinetic parameters and in vitro fertilization cycle outcomes.

PURPOSE: Although the impact of the paternal contribution to embryo quality and blastocyst formation is a well-known phenomenon, the current literature provides insufficient evidence that hyaluronan-binding sperm selection methods improve assisted reproductive treatment outcomes. Thus, we compared the cycle outcomes of morphologically selected intracytoplasmic sperm injection (ICSI) with hyaluronan binding physiological intracytoplasmic sperm injection (PICSI) cycles. METHODS: A total of 2415 ICSI and 400 PICSI procedures of 1630 patients who underwent in vitro fertilization cycles using a time-lapse monitoring system between 2014 and 2018 were analyzed retrospectively. Fertilization rate, embryo quality, clinical pregnancy rate, biochemical pregnancy rate and miscarriage rate were evaluated, differences in morphokinetic parameters and cycle outcomes were compared. RESULTS: In total, 85.8 and 14.2% of the whole cohort were fertilized with standard ICSI and PICSI, respectively. The proportion of fertilized oocytes did not significantly differ between groups (74.53 ± 1.33 vs. 72.92 ± 2.64, p > 0.05). Similarly, the proportion of good-quality embryos according to the time-lapse parameters and the clinical pregnancy rate did not significantly differ between groups (71.93 ± 4.21 vs. 71.33 ± 2.64, p > 0.05 and 45.55 ± 2.91 vs. 44.96 ± 1.25, p > 0.05). No statistically significant differences were found between groups in clinical pregnancy rates (45.55 ± 2.91 vs. 44.96 ± 1.25, p > 0.05). Biochemical pregnancy rates (11.24 ± 2.12 vs. 10.85 ± 1.83, p > 0.05) and miscarriage rates (24.89 ± 3.74 vs. 27.91 ± 4.91, p > 0.05) were not significantly different between groups. CONCLUSION: The effects of the PICSI procedure on fertilization rate, biochemical pregnancy rate, miscarriage rate, embryo quality, and clinical pregnancy outcomes were not superior. The effect of the PICSI procedure on embryo morphokinetics was not apparent when all parameters were considered.

The Effects of Microfluidic Sperm Sorting, Density Gradient and Swim-up Methods on Semen Oxidation Reduction Potential.

PURPOSE: To compare the effects of microfluidic sperm sorting, density gradient and swim-up methods on the oxidative reduction potential (ORP) of split semen samples from a single patient population. MATERIALS AND METHODS: A prospective controlled study was conducted to compare the effects of three different semen processing methods using split semen samples from the same population of infertile men. The primary outcome was the ORP. Secondary outcomes were the sperm concentration, progressive motility rate and total sperm motility. RESULTS: A total of 57 split semen samples were included in this study. The ORP was significantly lower in the microfluidic group compared to the density gradient and swim-up groups (p < 0.05). The ORP/sperm concentration ratio was significantly lower in the microfluidic and density gradient groups compared to the swim-up group (p < 0.05).Total sperm concentration was significantly higher in the density gradient group than the microfluidic and swim-up groups (p < 0.05). Motility was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05). The progressive motile sperm rate was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05). CONCLUSIONS: Microfluidic sperm sorting was better for selecting highly motile sperm and yielded a lower ORP than conventional sperm preparation methods.

Comparison of microfluid sperm sorting chip and density gradient methods for use in intrauterine insemination cycles.

OBJECTIVE: To compare the effect of microfluiding sperm sorting chip and density gradient methods on ongoing pregnancy rates (PRs) of patients undergoing IUI. DESIGN: Retrospective cohort study. SETTING: Hospital IVF unit. PATIENT(S): Couples with infertility undergoing IUI cycles between 2017 and 2018. INTERVENTION(S): Not applicable. MAIN OUTCOME MEASURE(S): Ongoing PRs. RESULT(S): A total of 265 patients were included in the study. Microfluid sperm sorting and density gradient were used to prepare sperm in 133 and 132 patients, respectively. Baseline spermiogram parameters, including volume, concentration, motility, and morphology, were similar between the two groups. Total motile sperm count was lower in the microfluiding sperm sorting group at baseline (35.96 ± 37.69 vs. 70.66 ± 61.65). After sperm preparation sperm motility was higher in the microfluid group (96.34 ± 7.29 vs. 84.42 ± 10.87). Pregnancy rates were 18.04% in the microfluid group and 15.15% in the density gradient group, and ongoing PRs were 15.03% and 9.09%, respectively. After using multivariable logistic regression and controling for confounding factors, there was a significant increase in ongoing PRs in the microfluid sperm sorting group. The adjusted odds ratio for ongoing pregnancy in the microfluid group compared with the density gradient group was 3.49 (95% confidence interval 1.12-10.89). CONCLUSION(S): The microfluid sperm sorting method significantly increased the ongoing PRs compared with the density gradient group in IUI cycles.

The use of in vitro maturation in stimulated antagonist in vitro fertilization cycles of normo-hyperresponder women due to arrested follicular development: A rescue procedure.

Objective: To evaluate the impact of rescue in vitro maturation (IVM) on the clinical outcomes of women with arrested follicular development in stimulated in vitro fertilization (IVF) cycles. Materials and Methods: This is a retrospective review of 13 patients who were evaluated as normo-hyperresponders for ovarian stimulation. The main outcome measure was the clinical pregnancy and livebirth rates. The purpose of gonadotropin stimulation in patients undergoing IVF is to retrieve multiple oocytes by avoiding multifetal gestation and Ovarian Hyperstimulation syndrome (OHSS). The ovarian response to stimulation ranges from poor response to OHSS, which is related to the follicular number and the dose of the gonadotropins used. However, in some cycles of normo-hyperresponder women, follicular development decelerates or ceases. Close follow-up in a daily manner and increasing the dose of gonadotropins did not change the follicular arrest. This clinical situation has two edges; one is cycle cancellation, which has undesired psychological outcomes for women and the IVF team, and second one is the prolongation of the IVF cycle. For such circumstances, IVM may be a valuable option. Stimulated IVF cycles were converted to IVM as a rescue IVM procedure following detailed informed consent of the women who were close to cycle cancellation. Results: Thirteen 13 IVM cycles and their clinical outcomes are presented. Six women achieved pregnancies, but only 4 delivered 5 healthy live born. The other two women had biochemical loss during follow-up. Conclusion: Based on the data obtained, it can be concluded that gonadotropin-stimulated cycles with follicular arrest at the edge of cancellation can be shifted to rescue IVM procedures with reasonable clinical outcomes.

Differences in Poly(ADP-ribose) Polymerase1- (PARP1-) and Proliferative Cell Nuclear Antigen (PCNA) Immunoreactivity in Patients Who Experienced Successful and Unsuccessful Microdissection Testicular Sperm Extraction Procedures.

PURPOSE: The aim of this study is to evaluate expression of deoxyribonucleic acid (DNA) synthesis and repair markers in testicular tissues of azoospermic men in whom sperm retrieval could and could not be achieved as a result of microdissection testicular sperm extraction (micro-TESE) procedure. MATERIALS AND METHODS: In this prospective cohort study, testicular tissues were retrieved from 60 Non-obstructive Azoospermia (NOA) patients who underwent micro-TESE procedure. These patients were divided into twogroups: micro-TESE positive group, which included 30 NOA patients from whom sperm could be extracted via micro-TESE procedure; and micro-TESE negative group, which included 30 NOA patients from whom sperm retrieval could not be achieved via micro-TESE procedure. Expression and distribution patterns of poly(ADP-ribose) polymerase-1 (PARP-1) and proliferative cell nuclear antigen (PCNA) in extracted tissues were assessedby immunohistochemical staining to reveal any differences in DNA synthesis and repair between the two groups. RESULTS: Micro-TESE positive group exhibited significantly stronger immunoreactivity for both PCNA and PARP-1 (P = .001 and P = .001 respectively). The results of this study reveal that both DNA synthesis and repair markers were expressed strongly in patients who experienced successful micro-TESE procedure. CONCLUSION: Although further studies are needed to support these findings, PARP-1 and PCNA expression in testicular tissues of NOA patients could be promising predictive factors for micro-TESE procedure success.

Pharmacokinetic, pharmacodynamic, and clinical aspects of ovulation induction agents: A review of the literature.

Controlled ovarian hyperstimulation is a key step for successful outcomes of assisted reproductive technique cycle outcomes. Many medications are available, which are commonly useed solely or in combination to achieve multiple follicular development. Pharmacokinetic, pharmacodynamic, and clinical information of ovulation induction drugs deserve to be elucidated for every individual patient before commencing infertility treatment. New concepts and new treatment protocols are introduced as ovulation physiology is understood by infertility specialists. Increasing treatment success by minimizing aderse effects is a milestone of all ovarian stimulation protocols that use these novel interventions. Achievement of a satisfactory cycle outcome includes retrieval of sufficient oocytes, a single clinical pregnancy, and avoidance of ovarian hyperstimulation syndrome. In this review, we evaluate the current literature to determine the most reliable and relevant information about the most used ovulation induction drugs.

Comparison of the morphokinetic parameters of embryos according to ovarian reserve in IVF cycles.

The aim of this study is to evaluate the impact of ovarian reserve and age of women on early morphokinetic parameters of embryos with a time-lapse monitoring system. In total, 197 infertile couples with poor ovarian reserve (Group 1, n = 41), normal ovarian reserve (Group 2, n = 59), or polycystic ovaries (Group 3, n = 97) were included. The time from insemination to the following events were analyzed: pronuclear fading (Pnf) and cleavage to 2, 3, 4 and 5 cells. The optimal ranges for morphokinetic parameters of t5, s2 and cc2 in each group were also evaluated. In total, 1144 embryos were evaluated. Morphokinetic parameters did not differ statistically between the groups. Data were analyzed according to different age groups (20-30, 30-40, >40). The morphokinetic parameters did not differ statistically in Group 1 and 3. In Group 2, the times from insemination to tPnf, t2, t3, t4 were significantly shorter in the younger age group than the older age group (p < 0.05). The percentages of optimal embryos, according to t5, s2 and cc2, did not differ statistically between the groups. In conclusion, ovarian reserve did not seem to affect the morphokinetic parameters of embryos.

The association between coenzyme Q10 concentrations in follicular fluid with embryo morphokinetics and pregnancy rate in assisted reproductive techniques.

PURPOSE: This study seeks to evaluate the association between follicular fluid (FF) coenzyme Q10 (CoQ10) levels, embryo morphokinetics, and pregnancy rate. METHODS: Sixty infertile patients who underwent intracytoplasmic sperm injection (ICSI) cycles were included in the study. For each patient, CoQ10 level of the follicular fluid was measured by high-performance liquid chromatography system. After the ICSI of each oocyte, the relationship between the level of CoQ10 content of each follicular fluid, the subsequent embryo quality, and embryo morphokinetics was investigated. The relationship between the level of CoQ10 content of each follicle and optimal time-lapse parameters for the embryos of these follicles including t5, s2, and cc2 was also analyzed. The embryos were further classified into four categories, namely, grades A, B, C, and D, according to morphokinetic parameters using t5-t2 and t5-t3 (cc3). Each follicular fluid analysis was performed for a single oocyte of a single embryo which was transferred to the patients. Additionally, follicular fluid CoQ10 levels and pregnancy rates were evaluated. RESULTS: Follicular fluid CoQ10 levels were significantly higher in grades A and B than grades C and D embryos (p < 0.05). The concentration of CoQ10 levels was significantly higher in the pregnant group (p < 0.05). There was no significant correlation between optimal t5 and s2 morphokinetic parameters and CoQ10 levels. However, CoQ10 levels were significantly higher in follicular fluid of embryos which had optimal cc2 (p < 0.05). CONCLUSION: High follicular fluid CoQ10 level is associated with optimal embryo morphokinetic parameters and higher pregnancy rates.

Comparison of corifollitropin alfa and daily recombinant follicle-stimulating hormone in poor responder patients undergoing in vitro fertilization cycles.

OBJECTIVE: The aim of this study was to compare the effect of corifollitropin alfa (CFA) and recombinant follicle-stimulating hormone (rFSH) in poor-responder patients undergoing antagonist cycles. MATERIALS AND METHODS: The study was a retrospective analysis of the treatment results of 214 poor responder patients who had been admitted to the In Vitro Fertilization Unit of Izmir Medical Park Hospital between November 2014 and November 2016. Intracytoplasmic sperm injections were performed in 38 patients (group 1) with CFA, and the remaining 176 (group 2) with rFSH for controlled ovarian hyperstimulation. RESULTS: The age, body mass index, anti-müllerian hormone level, duration of infertility, duration of induction and antral follicle number were similar in the two groups. There was no difference in the total aspirated oocyte counts, mature oocyte ratio, fertilization rate, implantation rate, and clinical pregnancy rates between the two groups. The implantation rate was 9/38 (23.6%) in group 1 and 42/176 (23.8%) in group 2, whereas the clinical pregnancy rates were 16.3% and 17.2%, respectively. CONCLUSION: No difference was found in terms of oocyte count, fertilization rate, implantation rate, and clinical pregnancy rates of CFA or rFSH use in the antagonist cycles in poor-responder patients.

Gonadotropin-releasing hormone agonist triggering is effective, even at a low dose, for final oocyte maturation in ART cycles: Case series.

OBJECTIVE: To investigate the efficacy of low-dose gonadotropin-releasing hormone (GnRH) agonist for final oocyte maturation in females undergoing assisted reproductive treatment (ART) cycles. MATERIAL AND METHODS: Nine females undergoing ovarian stimulation in a GnRH antagonist protocol who received triptorelin 0.1 mg to trigger final oocyte maturation were included. Treatment outcomes of these patients were compared with those of controls, matched for age and oocyte number (n=14), who received 0.2 mg triptorelin at the same time. The luteal phase was supported with vaginal micronized progesterone and oral estradiol hemihydrate 2 mg twice daily. RESULTS: The mean (±) numbers of retrieved, metaphase II, and fertilized oocytes were 15.66±7.82, 14±7.28, and 10.11±5.86, respectively. The implantation and clinical pregnancy rates were 46.1% and 71.4%, respectively. Of the pregnancies, 2 were live births, 1 was a preterm birth (twins), 2 are on-going, and 2 ended as miscarriages. No case of OHSS was encountered. On comparison of the results of these patients (fresh cycles; n=7) with those of matched controls, there were no significant differences in terms of retrieved mature oocytes, implantation rates, or clinical pregnancy rates (p>0.05). CONCLUSION: These findings suggest that low-dose GnRH agonist triggering has similar efficacy as standard doses in terms of retrieved mature oocytes and clinical pregnancy rates in in vitro fertilization cycles.

The combination of dehydroepiandrosterone, transdermal testosterone, and growth hormone as an adjuvant therapy in assisted reproductive technology cycles in patients aged below 40 years with diminished ovarian reserve.

OBJECTIVE: To evaluate to the efficacy of testosterone, dehydroepiandrosterone (DHEA) and growth hormone (GH) supplementations in patients with diminished ovarian reserve (DOR) in assisted reproductive technology (ART) cycles. MATERIALS AND METHODS: A retrospective cohort including 33 women with 81 ART cycles were aged and ovarian reserve matched 52 women with 102 conventional in vitro fertilization (IVF)/intra-cytoplasmic sperm injection (ICSI) protocol. Administration of DHEA for 12 weeks and transdermal testosterone for 4 weeks as pretreatment adjuvant and luteal start GH in DOR patient treatment arm compared to conventional IVF/ICSI cycles. RESULTS: The number of follicles >14 mm, number of oocytes, number of metaphase 2 oocytes and fertilisation rate were significantly higher in ISIK protocol (IP). The clinical pregnancy rate (CPR) per embryo transfer of the IP was 38.2% (13/34). The cancellation rate of cycles decreased significantly from 54.5 % (24/44) to 8.1% (3/37) with the IP, while the OPR was 35.3% (12/34). CONCLUSIONS: Our study has shown that even the poorest responders could achieve clinical pregnancy after inducing ovarian folliculogenesis with a combination of transdermal testosterone, DHEA and GH.

Gonadotrophin-releasing hormone agonist trigger and freeze-all strategy does not prevent severe ovarian hyperstimulation syndrome: a report of three cases.

Ovarian hyperstimulation syndrome (OHSS) is the most serious iatrogenic complication of IVF cycles. Although the development of effective treatment strategies for this syndrome is important, preventing OHSS is more crucial. Triggering ovulation with a gonadotrophin-releasing hormone (GnRH) agonist is one method used to avoid OHSS. In this paper, three patients who developed severe OHSS after undergoing GnRH agonist triggering and freezing of all embryos in a GnRH antagonist protocol are described. A review of the literature is also provided. This report highlights the ongoing risk of severe OHSS even after GnRH agonist triggering combined with freezing all embryos in GnRH antagonist cycles. Other prevention strategies might be considered for extreme hyper-responders.

Endometrial spiral artery Doppler parameters in unexplained infertility patients: is endometrial perfusion an important factor in the etiopathogenesis?

OBJECTIVE: Uterine perfusion, particularly the endometrial blood flow, may have an important role in endometrial receptivity. In order to assess the contribution of sub endometrial blood flow in the etiopathogenesis of unexplained infertility mid luteal- peri-implantation period spiral artery transvaginal color Doppler parameters were measured and compared with fertile controls. MATERIAL AND METHODS: Forty-two consecutive patients admitted to Izmir Katip Celebi University Ataturk Training and Research Hospital, Department of Obstetric and Gynecology with the diagnosis of unexplained infertility after standard diagnostic work up constituted the study group and they were compared with a fertile control group admitted to hospital with non specific gynecological complaints or for check-up in the same period. Mid luteal transvaginal color Doppler ultrasonography was applied to each patient by the same radiologist who was blind to the diagnosis of the particular patient and, RI (resistance index) and PI (pulsatility index) values were calculated. RESULTS: There were no significant differences between the two groups, in respect to age, body mass index, basal hormonal and mid luteal progesterone levels (p>0.05). For the fertile control group, mid luteal-peri-implantation phase endometrial spiral artery mean RI values were calculated as 0.48±0.08 SD and mean PI values as 0.65±0.18 SD. For the study group, mean RI values were calculated as 0.54±0.07 SD, PI values were calculated as 0.80±0.16 SD. The differences for RI (p=0.009) and PI (p=0.004) were statistically significant. CONCLUSION: According to Doppler parameters, unexplained infertility patients have high impedance blood flow in spiral arteries which means that peri-implantation blood flow in these patient is lower than fertile controls. These findings suggest that endometrial perfusion may have an important contribution to etiopathogenesis of unexplained infertility.

The effect of retained embryos on pregnancy outcome in an in vitro fertilization and embryo transfer program.

OBJECTIVE: The purpose of this study was to determine the incidence of retained embryos and its impact on pregnancy outcome in the absence of known risk factors like blood and mucus in the transfer catheter. The factors that could be associated with embryo retention were also investigated. STUDY DESIGN: The results of all embryo transfer procedures performed at Ankara IVF Center between January 2003 and December 2005 were analyzed retrospectively. Three hundred and five embryo transfers, in which the transfer catheter was contaminated with blood or mucus, were excluded and the remaining 1,454 embryo transfers, with clean catheter, were enrolled into the study. Both fresh (n=1,422) and frozen (n=32) embryo transfers were included. RESULTS: The overall incidence of retained embryos during study period was 2.8% (41/1,454) following a clean initial embryo transfer. The mean age of the female partner, mean number of retrieved oocytes, MII oocytes, fertilized oocytes and the embryos transferred were similar in patients with and without retained embryos. The 1,454 embryo transfers performed during the study period resulted in 712 pregnancies (49%), of which 639 proved to be clinical pregnancies with a rate of 44%. The implantation rate was 22.8%. Pregnancy outcomes including positive beta-hCG (58.5% versus 48.7%), biochemical (4.7% versus 5.1%) and clinical pregnancy rates (53.6% versus 43.6%), implantation rate (24% versus 22.7%) and multiple pregnancy rate (36.3% versus 44.7%) were not significantly different between patients with and without retained embryos. An influence of individual physicians and embryologists on the frequency of retained embryos was not detected. The type of embryo transfer catheter used did not show any difference in terms of embryo retention. The cleavage stages of embryos were not different between groups. Although it was not statistically significant, the number of embryos transferred appeared to be a potentially confounding factor for retained embryos (p=0.053) and it might be significant in a slightly larger sample. When transfer of one or two embryos was compared to three or more embryos, the likelihood of retained embryos increased from 1.2% (4/321) to 3.2% (37/1,133). CONCLUSION: Retained embryos in the transfer catheter and immediate retransfer of them have no adverse impact on clinical pregnancy and implantation rates unless other previously reported signs of difficult transfer are also observed.

Genetic aspects of human male infertility: the frequency of chromosomal abnormalities and Y chromosome microdeletions in severe male factor infertility.

OBJECTIVE: The main purpose of this study is to detect the frequency and type of both chromosomal abnormalities and Y chromosome microdeletions in patients with severe male factor infertility and fertile control subjects. The association between the genetic abnormality and clinical parameters was also evaluated. METHODS: This study was carried out in 208 infertile and 20 fertile men. Results of 208 patients, 119 had non-obstructive azoospermia and 89 had severe oligoasthenoteratozoospermia (OAT). Seventeen out of 119 (14.3%) azoospermic patients and two out of 89 (2.2%) patients with OAT had Y chromosome microdeletions. In total, 19 cases with deletions were detected in 208 infertile men, with a frequency of 9.1%. The AZFc locus, mainly DAZ gene cluster was the most frequently deleted region. Five other cases with azoospermia (4.2%) and two cases with OAT (2.2%) had a chromosomal abnormality, with a total number of seven (3.4%). Including Y chromosome deletions and structural chromosome abnormalities, the rate of genetic abnormalities was 12.5% (26/208) in our patients. On the other hand, 20 men with proven fertility and fathers of five cases with microdeletions were genetically normal. Y chromosome deletions and chromosomal abnormalities were associated with various histological alterations in testis. Sertoli cell-only (SCO) syndrome and maturation arrest predominated in these cases, whereas hypospermatogenesis occurred more frequently in genetically normal patients. CONCLUSION: Various chromosomal abnormalities and deletions of Y chromosome can cause spermatogenic breakdown resulting in chromosomally derived infertility. All these findings strongly support the recommendation of genetic screening of infertile patients.