Comparison of different stimulation protocols efficacy in poor responders undergoing IVF: A retrospective study

Purpose: To compare the efficacy of different stimulation protocols on pregnancy outcomes in poor responders undergoing in vitro fertilization (IVF). Materials and methods: This was a retrospective study to compare the efficacy of four different protocols including gonadotropin-releasing hormone (GnRH) agonist (long, short and miniflare) and GnRH antagonist on pregnancy outcomes in poor responders. This investigation was performed on 566 poor respond patients who were candidates for IVF. Main outcome measures included the total number of oocytes and mature oocytes retrieved, pregnancy rates, implantation and overall cancellation rates which were compared between four mentioned groups. Results: Number of follicles >18?mm on hCG day were significantly higher in GnRH-a long versus GnRH antagonist, GnRH-a short and GnRH-a miniflare protocols. The mean number of oocytes and mature oocytes retrieved were significantly higher in GnRH-a long versus miniflare (4.7?±?3.05 versus 3.26?±?2.9 and 3.69?±?3.1 versus 2.65?±?2.2, respectively). There were no significant differences in implantation, pregnancy and overall cancellation rates between four groups. Conclusion: The present study suggests that the application of four different protocols in poor respond patients seem to have similar efficacy in improving clinical outcomes such as implantation, pregnancy rates and cancellation rate even though GnRH-a long protocol yielded more retrieved oocytes and mature oocytes compared to GnRH-a miniflare protocol.     

Is there a benefit from routine follicular flushing for oocyte retrieval?

The object of this study was to determine the optimum number of follicular flushing for a maximum number of oocytes to be retrieved. This was a prospective study, based at a private in vitro fertilization centre (Brentwood Fertility Centre, The Essex Nuffield Hospital, Brentwood, Essex UK). Patients attending the IVF centre for assisted reproductive technology were studied. We counted the number of oocytes obtained with each flushing after primary aspiration of the follicle. Some 40% of the oocytes were retrieved with primary aspiration without follicular flushing, while up to 82% of oocytes were retrieved with two flushes and 97% of oocytes were retrieved in up to four flushes. Only 3% of the remaining oocytes were retrieved with 5th and 6th flush. The optimum number of follicular flushing was four times, in order to achieve maximum number of oocytes without compromising the duration of retrieval and number of oocytes retrieved.     

卵泡发育迟缓者在体外受精中延长使用促性腺激素的临床疗效观察

目的探讨体外受精（IVF）周期中,卵泡发育迟缓者延长使用促性腺激素（Gn）的临床效果。方法将552个IVF周期按Gn用药时间、剂量和获卵数分组,观察组（69个取卵周期,66个移植周期）：Gn起始用量为75IU／d-300IU／d,用药时间≥16d,用至卵泡达到取卵标准为止,获卵数≥4个;对照组（483个取卵周期,464个移植周期）：Gn起始用量为150IU／d～300IU／d,用药时间≤15d,且总Gn用量≤3300IU,获卵数≥4个。比较两组患者的Gn用量及用药时间、获卵数、临床妊娠率、胚胎着床率及治疗结局。结果观察组和对照组的临床妊娠率分别为45．5％（30／66）和51．7％（240／464）;胚胎着床率分别为28．0％（46／164）和30．5％（385／1262）;分娩率分别为37．9％（25／66）和39．4％（183／464）;两组患者的临床妊娠率、胚胎着床率、分娩率等比较,差异均无统计学意义（P〉0．05）。观察组和对照组的多囊卵巢（PCO）和（或）多囊卵巢综合征（PCOS）患者的比例分别为55．1％（38／69）和20．1％（97／483）;基础窦卵泡数分别为（20±11）个和（15±6）个;Gn用药时间分别为（20．8±4．2）d和（10．3±1．8）d;Gn用药总量分别为（3090±1140）IU和（2302±862）IU;血清雌二醇峰值分别为（4595±5181）pmol／L和（7272±6320）pmol／L;获卵数分别为（10±6）个和（14±7）个;移植胚胎数分别为（2．5±0．6）个和（2．7±0．5）个;两组上述各指标比较,差异均有统计学意义（P〈0．01）。结论对IVF周期中卵泡发育迟缓的患者,延长使用Gn有效、方便,能获得满意的临床结局。     

Does follicular flushing increase oocyte number in poor responders? An update of a systematic review

Research questionDoes flushing of the follicles at ovum retrieval increase the number of retrieved oocytes in poor-response IVF patients?DesignAn update of an electronic literature search was performed to identify randomized controlled trials (RCT) investigating follicular flushing versus no flushing in women with a poor response to IVF treatment. No exclusion criteria for type of needle, stimulation or protocol were applied. A meta-analysis was conducted using the software RevMan 5.4.ResultsSix RCT were identified that had the primary objective of testing for an increase in mean number of cumulus–oocyte complexes or/and metaphase II oocytes between flushing and no flushing. A double-lumen needle was used in five trials, one study investigated a pseudo-double-lumen needle, and a conventional single-lumen needle was used in all the control groups. The main risk of bias in all the included studies is a lack of blinding of the physicians performing the puncture and incomplete data in four trials. A heterogeneity of direction and size of effect of follicular flushing on mean oocyte number retrieved was detected (I² = 80, P = 0.0001), which precludes a synthesis of the data. Two studies showed a decrease or tendency towards a decrease in oocyte numbers, one study showed similar oocyte numbers, and one study showed a strong tendency towards increased oocyte numbers with flushing. A similar picture was seen for metaphase II oocytes (I² = 73, P = 0.002).ConclusionsIt is uncertain whether follicular flushing in poor-response IVF patients affects the number of retrieved oocytes. Larger pragmatic trials are warranted to clarify the effect of flushing on oocyte numbers and clinical outcomes in poor responders and monofollicular patients.     

The follicular flushing duplicate the pregnancy rate on semi natural cycle IVF

Objective

To evaluate the importance of follicular flushing on semi natural cycle IVF.

Material and methods

We have compared prospectively the reproductive potential of oocytes obtained from follicular fluid (LF, N  =  79) to those obtained from follicular flushing (R, N  =  47) in 146 oocyte pick ups.

Results

The group LF and R were similar with regard to fertilization rate (79.7 versus 88.1%, respectively), percentage of superior grade embryos (28.8 versus 37.8%) and implantation rate (24.1 versus 44.1%).

Conclusion

The practice of follicular flushing on semi natural IVF cycle improves the pregnancy rate. The oocytes obtained by follicular flushing had the same reproductive potential than those obtained on follicular fluid.     

Age-matched comparison of recombinant and urinary HCG for final follicular maturation

This age-matched retrospective analysis compared the clinical outcomes of recombinant human chorionic gonadotrophin (rHCG) and urinary HCG (uHCG) in patients undergoing fresh, nondonor IVF cycles. The patients underwent ovarian stimulation by standard gonadotrophin-releasing hormone (GnRH) agonist down-regulation or a GnRH antagonist protocol using recombinant FSH (rFSH) alone or in combination with human menopausal gonadotrophin. When two or more follicles had attained a mean diameter of 20 mm, follicular triggering was achieved with either Ovidrel (rHCG) 250 mug SC or uHCG 10,000 IU IM. Patients receiving rHCG were considered subjects, and they were age-matched in a 1:2 ratio to patients receiving uHCG, who were designated as controls. The main outcome measures were number of oocytes retrieved, number of mature oocytes obtained, number of oocytes fertilized and clinical pregnancy rates. A total of 273 subjects were age-matched and compared with 546 controls. Recombinant HCG had a minimal effect on the number of oocytes retrieved (13.4 versus 13.2), mature oocytes (10.5 versus 10.3) and oocytes fertilized (8.2 versus 7.8) compared with uHCG. Pregnancy (46.0 versus 45.2%) and clinical pregnancy rates (38.1 versus 36.8%) were similar for rHCG and uHCG. Recombinant HCG was as effective as uHCG for final follicular maturation in IVF cycles.     

A new method to assess oxidative stress in ART cycles

The role of oxidative stress (OS) in female reproduction is an understudied area for investigation. The aim of the study was to assess reactive oxygen species (ROS) levels both in follicular fluid (FF) and in serum samples of women undergoing IVF and to relate them to clinical outcomes. A total of 56 women were enrolled. From each patient a sample of serum and FF were collected at oocyte retrieval day and analyzed for OS, by measuring a class of reactive oxygen metabolites (ROMs) (dROMs test) and for biological anti-oxidant potential levels (BAP test). The data analyzed in serum and in FF were related to pregnancy outcomes. There was no significant relationship between d-ROMs and BAP levels in serum or FF GROUP A and B and the clinical parameters of prognosis of women: age, number of oocytes retrieved number of oocytes methaphase II, FSH, fertilization rate and cleavage rate, number of embryos transferred and embryo score. There was a positive association between systemic levels of OS measured in serum GROUP A and pregnancy outcomes (p?=?0.007) suggesting that the evaluation of OS in FF needs more investigation about markers in follicular microenvironment in order to predict IVF success.     

卵巢高反应对体外受精-胚胎移植的影响

目的探讨体外受精-胚胎移植周期控制性超排卵（COH）中卵巢高反应对妊娠结局的影响。方法回顾性分析中山大学附属第二医院生殖中心1082个IVF／ICSI周期的临床资料，根据HCG注射日血E2〉11010pmol／L或获卵数〉15个定义为高反应组，HCG注射日血E2〉1835pmol／L，且获卵数5～15个为正常反应组，比较两组的妊娠结局。结果与正常反应组相比，高反应组获得的优质胚胎数显著增多（P〈0．001），但两者的受精率、卵裂率、胚胎种植率、临床妊娠率、流产率比较，差异无显著性（P〉0．05）。结论COH中卵巢高反应对妊娠结局无明显影响，但需注意预防卵巢过度刺激综合征的发生.     

Effect of aspiration vacuum on the developmental competence of immature human oocytes retrieved using a 20-gauge needle

In-vitro maturation (IVM) of immature oocytes has been proposed as a potential alternative to conventional IVF treatment following ovarian stimulation. However, the effects of the oocyte retrieval conditions on subsequent development have not been well understood. This study assessed the effects of different aspiration vacuums during oocyte retrieval on the developmental competence of immature oocytes following IVM, IVF and embryo transfer, retrospectively. Immature oocytes were aspirated with 20-gauge needles with a vacuum of 180 or 300 mmHg. Immature oocytes were cultured in IVM medium for 26 h. All mature oocytes were inseminated by intracytoplasmic sperm injection (ICSI). Embryo transfer was carried out 2 or 3 days after ICSI. The percentage of cumulus-cell enclosed oocytes and of transferable embryos per retrieved oocytes in 180 mmHg (69.7% and 23.8%, respectively) were significantly higher (P < 0.01) than those in 300 mmHg (46.2% and 12.8%, respectively). The ongoing pregnancy rate per retrieval cycle in 180 mmHg (30%) was higher (P < 0.01) than that in 300 mmHg (4.3%). The data indicate that lower pressure of vacuum aspiration with a 20-gauge needle improves the developmental competence of immature oocytes following IVM, IVF and embryo transfer.     

Conventional IVF as a laboratory strategy to rescue fertility potential in severe poor responder patients: the impact of reproductive aging

Abstract

Objective: To investigate whether laboratory strategies can improve in vitro fertilization (IVF) outcome in poor responder patients. We compared the effectiveness of conventional IVF and intra cytoplasmic sperm injection (ICSI) in assisted reproductive technologies cycles in which only one or two oocytes were retrieved at ovarian pick up, in the absence of male infertility.

Design: Retrospective analysis of 425 cycles in 386 poor responder patients.

Intervention(s): Standard stimulation protocol with gonadotropins and gonadotropin releasing hormone (GnRH) antagonist.

Main outcome measure(s): Fertilization rate, cleavage rate, good-quality embryo rate, implantation rate, clinical pregnancy rate (PR) and miscarriage rate.

Results: IVF was found to be more advantageous for implantation and PR, especially in patients under 35 years and in women aged between 35 and 38 years. No differences were noted in the other parameter evaluated. Patients aged over 38 years showed no difference using the two techniques.

Conclusion: The employment of ICSI in the absence of a male factor can reduce reproductive outcome in poor responder. Probably because of aging-related defects overcoming the advantage of sperm selection, the choice of IVF technique is not relevant to reproductive success when oocyte quality is compromised by reproductive aging. Although further randomized trials are needed to confirm our results, we propose that, in absence of male infertility, conventional IVF might be the technique of choice in young patients, especially in those aged below 35 years.     

Correlation between follicular diameters and flushing versus no flushing on oocyte maturity,fertilization rate and embryo quality

Objective

To determine (a) the correlation between follicular sizes, oocyte maturity, normal fertilization rate, cleavage and embryo quality; and (b) to establish whether oocytes recovered with or without follicular flushing have different developmental competence.

Design

Prospective observational study.

Setting

Academic medical center.

Patients

Forty nine cycles (37 ICSI and 12 IVF).

Interventions

Measurement of 360 follicular diameters on the day of egg retrieval and classification into three groups Group A (mean diameter 12–14.5 mm.), group B (mean diameter 15–18 mm.) and group C (diameter >18.5 mm.).

Main outcome measure

Correlation between follicular size at the time of retrieval and oocyte maturity, fertilization and cleavage rate in 226 oocytes (163 ICSI and 63 IVF). Developmental competence of oocytes retrieved with flushing versus non flushing.

Results

Almost all (99 %) of the oocytes recovered from follicles of group C were in metaphase II as opposed to 80 % in group A and 81 % in group B (p < 0.01). Overall there was a progressive and significant increase in fertilization rates from group A follicles to group C (47 % vs. 67 %, p 0.05). Overall 53 % of oocytes retrieved from group A follicles showed either no fertilization or abnormal fertilization versus 27 % in group C (p 0.05). The oocyte recovery rate with follicular flushing improved from group A to group B and to group C follicles (65 % vs. 49 % vs.37 % respectively p < 0.01). There were no differences in rates of immature oocyte, fertilization, abnormal or not fertilization and cleavage.

Conclusions

The results of this study shows that: a) Follicles larger than 18 mm at retrieval have consistently mature oocytes with a higher rate of fertilization; b) Small size follicles are still capable of containing mature oocytes, but their rate of abnormal or no fertilization is high; c) Oocytes recovered with flushing are still able to produce embryos with full developmental competence.     

Dual stimulation using corifollitropin alfa in 54 Bologna criteria poor ovarian responders – a case series

Research question

What are the reproductive outcomes of Bologna criteria poor responders undergoing dual stimulation (DuoStim) and subsequent cryopreserved embryo transfer?

Effect of follicular flushing on reproductive outcomes in patients with poor ovarian response undergoing assisted reproductive technology

Purpose

The purpose of this study is to investigate the impact of follicular flushing on the number of oocytes retrieved, oocyte maturity, fertilization rate, embryo development, and pregnancy rate of poor ovarian responders (POR).

Methods

Retrospective study of 524 cycles of 384 patients with POR submitted to assisted reproductive technology (ART) and who had follicular flushing during oocyte retrieval was used in the study. We included patients with <5 oocytes at oocyte retrieval (POR group) and matching the Bologna criteria.

Results

POR patients had a mean age of 38.2 ± 4.2 years. A total of 1355 follicles (mean = 3.5 ± 1.6) were aspirated and 1040 oocytes recovered, with 709 (68.2%) obtained by direct aspiration and 331 (31.8%) by follicular flushing. We found a difference between the total number of oocytes and the number of aspirated oocytes. Overall pregnancy rate was 22%. Association was observed between pregnancy rate and the number of oocytes retrieved, the number of MII oocytes, and the number of embryos transferred. The patients matching the Bologna criteria had a mean age of 38.9 ± 3.9 years. A total of 309 follicles were aspirated (mean = 3.1 ± 1.5) and 242 oocytes recovered, with 156 (64.5%) obtained by direct aspiration and 86 (35.5%) by follicular flushing. There was a significant difference between the total number of oocytes and the number of aspirated oocytes. Overall pregnancy rate was 12.1%. There was no association between the pregnancy rate and the number of oocytes retrieved, the number of MII, and the number of embryos.

Conclusions

Follicular flushing might be a suitable alternative to increase the number of oocytes and pregnancy rates in patients with POR.

     

Leptin,VEGF, IGF-1, and IGFBP-3 concentrations in serum and follicular fluid of women undergoing in vitro fertilization

We studied the influence of recombinant follicle-stimulating hormone (rFSH) stimulation on the concentration of leptin, vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) in serum and follicular fluid (FF) in women undergoing assisted reproduction. To test the hypothesis that these hormones could predict successful implantation and that the levels correlate with pregnancy rate. Sequential serum samples were drawn at the beginning of stimulation and on the day of embryo transfer (ET) from 84 women undergoing IVF. The follicular fluid (FF) obtained during oocyte retrieval was collected and the concentration of leptin, VEGF, IGF-1 and IGFBP-3 were measured in all samples. The hormones were measured by commercially available IRMA, RIA or EIA. Patients' characteristics and results of the assisted reproductive cycles were registered. Serum concentrations of VEGF, IGF-1 and IGFBP-3 significantly decreased during rFSH treatment. In contrast, serum leptin significantly increased after rFSH treatment. A strong correlation was found between the FF - levels of IGF-1, IGFBP-3, leptin and respective serum levels. With regard to IVF outcome, higher serum concentrations of IGF-1, IGFBP-3 and VEGF on the day of oocyte retrieval were observed in conception cycles vs. non-conception cycles. No such difference, however, was apparent at the beginning of the stimulation cycle. There was no association between FF levels of any of these hormones and IVF outcome. Our results demonstrate that VEGF, IGF-1, IGFBP-3 and leptin levels are affected by rFSH during controlled ovarian hyperstimulation and that there is a direct association between serum and FF levels, albeit without clinical implications     

Intrafollicular and serum levels of leptin during in vitro fertilization cycles: comparison between the effects of recombinant follicle-stimulating hormones and human menopausal gonadotrophin

Objectives.?To compare the effect of recombinant follicle-stimulating hormones (r-FSH) and human menopausal gonadotrophin (hMG) on leptin levels in serum and follicular fluid (FF) during in vitro fertilization IVF/ET treatment, and to investigate whether leptin levels in the follicular fluid and/or serum are correlated with IVF success.

Methods.?Sixty-three patients undergoing IVF cycle were subdivided into two groups. r-FSH was used to for controlled ovarian hyperstimulation in 29 patients (Group A) while, hMG was used in 34 patients (Group B). Our main outcomes were serum and FF leptin on the day of oocyte collection.

Result(s).?The two groups were comparable in age, body mass index (BMI), indications for IVF/ET, E2 level on human chorionic gonadotrophin day, number of retrieved oocytes, fertilization rate, number of transferred embryos and pregnancy rate. Serum and FF leptin levels were similar between the two study groups. Additionally, no correlation was found between levels of leptin in either serum or FF and cycle results such as: number of retrieved oocytes, fertilization rate and pregnancy rate.

Conclusions.?r-FSH and hMG have been found to have comparable effects on leptin levels in the serum and the FF of patients undergoing IVF/ET. Additionally, leptin levels in both serum and FF on day of retrieval have no correlation to IVF/ET outcome.     

Embryo implantation after human in vitro fertilization: importance of endometrial receptivity

To study the effects of controlled ovarian hyperstimulation (COH) on endometrial receptivity during human in vitro fertilization (IVF), we compared embryo implantation data obtained from our nonanonymous donor oocyte program with those obtained from standard IVF. To control for embryo quality, IVF patient characteristics were matched to those of the oocyte donors with regard to age and previous gravidity. All cycles were performed in an identical manner, using standardized ovarian stimulation protocols, transvaginal follicle aspiration, fertilization and culture in vitro, and transcervical embryo transfer. A similar number of oocytes per cycle were obtained, fertilization rates were similar, and similar numbers of embryos were transferred in both groups. Transferred embryos had similar numbers of blastomeres and were morphologically judged to be of similar quality in both groups. Implantation rates per individual embryo were significantly higher in donor than in standard IVF (35% versus 10.7%) as were ongoing per embryo implantation rates (23% versus 8%). Clinical and ongoing pregnancy rates per cycle were likewise higher in the donor group than in the standard IVF group (67% versus 39%, and 61% versus 30%, respectively). We conclude that COH inhibits embryo implantation after IVF by decreasing endometrial receptivity, which is an important factor in IVF pregnancy success.     

Impact of duration and dose of gonadotrophins on IVF outcomes

In order to examine whether the duration of the follicular phase and changes in daily gonadotrophin dosages impact IVF outcome, a retrospective analysis of women who underwent oocyte retrieval and fresh embryo transfer was performed. Among the parameters assessed were the number of days of gonadotrophin stimulation, changes in the daily dosage of gonadotrophins, total ampoules of gonadotrophins, embryo implantation rates, clinical pregnancy rates and ongoing pregnancy rates. The number of days of gonadotrophin stimulation, as determined by standard follicular size criteria did not appear to influence IVF outcomes. There was no significant difference in pregnancy rates between women who were stimulated for <9 days, 10-11 days or >12 days. When grouped by amount of starting daily dose of gonadotrophins there was a significant inverse relationship between gonadotrophin requirements and pregnancy rates (P=0.02). The data suggest that the success of an IVF cycle depends on the ovaries' ability to develop follicles of the appropriate size, not the speed at which the ovaries perform this function.     

Sibling embryo blastocyst development correlates with the in vitro fertilization day 3 embryo transfer pregnancy rate in patients under age 40

OBJECTIVE: To examine the IVF day 3-ET pregnancy rate in patients under 40 with sibling embryo blastocyst development, compared with similar patients without blastocyst formation. DESIGN: Retrospective analysis. SETTING: Academic infertility center. PATIENT(S): One hundred twenty-five IVF day 3-ET patients under 40 with sibling embryos for extended culture. INTERVENTION(S): Extended culture of nontransferred sibling embryos for blastocyst development. MAIN OUTCOME MEASURE(S): Pregnancy and multiple gestation rates, number of oocytes, embryos formed, and embryos transferred. RESULT(S): Thirty-eight percent of patients became pregnant. Forty-eight percent of patients had sibling embryos develop to blastocyst. The blastocyst group had more oocytes retrieved (17.4+/-6.6 versus 14.4+/-5.6), more embryos formed (11.2+/-4.2 versus 8.8+/-3.2), and a higher clinical pregnancy rate (60% versus 18%) than the group without blastocyst development. CONCLUSION(S): Blastocyst transfer has been shown to improve implantation rates and reduce the risk of multiple gestations from assisted reproductive technology. Sibling embryo blastocyst development may reflect superior embryo quality, as manifested by increased IVF-ET pregnancy rates. In addition to predicting pregnancy in the current cycle, sibling embryo blastocyst development may provide information about the potential for fresh blastocyst transfer in subsequent cycles and help to identify patients at risk for multiple gestations.     

Embryo quality based on ovulation induction: defining the differences

Patients undergoing IVF, with or without intracytoplasmic sperm injection, were treated with either recombinant human FSH or urine-derived FSH. Response to ovarian stimulation was monitored by ultrasound examinations and measurement of serum oestradiol concentrations. To define any differences in embryo quality and hence assisted reproductive technology success rates, a retrospective analysis of 811 recombinant FSH versus 555 urinary FSH cycles was undertaken. Embryo quality was assessed as embryo cell number and degree of fragmentation. Implantation and ongoing pregnancy rates were also compared. Use of recombinant FSH resulted in a higher percentage of mature oocytes, improved embryo cleavage, with more embryos available for freezing and higher implantation rates compared with urinary FSH. Oocyte and embryo quality were superior when recombinant FSH was used for ovarian stimulation compared with urinary FSH.