There is a relation between adipose tissue and macrophages. These macrophage cells influence adipose tissue by releasing many bioactive chemicals, including the wingless-type MMTV integration site family 5a (wnt5a) glycopeptides and the more traditional cytokines. Healthy adipocytes can release the wnt5a inhibitor Secreted frizzled-related protein 5(SFRP5). However, this protective effect was found to be reduced in obesity. This study aimed to assess the effect of follicular and serum levels of SFRP5 as well as serum and follicular levels of wnt5a on ICSI outcome and determine the potential roles of SFRP5 as a predictor of positive pregnancy in Iraqi infertile females undergoing ICSI with different body mass index .Ninety infertile females who received intracytoplasmic sperm injection and had various infertility factors between 18 and 45 years participated in this prospective clinical cross-sectional study. Follicular fluid, serum SFRP5, and serum and follicular fluid wnt5a levels were measured for all participants on the day of ovum pickup. In addition, ICSI outcomes were determined. There was a significant difference in the mean total gonadotrophin dose used among the three studied groups (p = 0.023), higher in overweight and obese females than normal-weight females. There was a positive significant correlation between serum plus follicular fluid wnt5a and total gonadotropin dose. Serum and follicular fluid SFRP5 levels were significantly higher in normal-weight females. There were higher serum (p-value=0.002) and follicular fluids (p-value=0.005) SFRP5 levels in pregnant females. Both SFRP5 and wnt5a glycoproteins were detected in infertile women's serum and follicular fluid. Serum and follicular fluid secreted frizzled-related protein 5 level positively associated with ICSI outcome. The cutoff values for serum and follicular fluid SFRP5 were ≥15.86 (ng/ml) and ≥ 11.3(ng/ml), respectively, as predictors of positive pregnancy in women undergoing IVF/ICSI cycles. Obesity, BMI, SFRP5, wnt5a, ICSI Outcome
Resistin and follistatin levels have been found to be high in polycystic ovarian syndrome (PCOS) patients, that play role in pathophysiology of PCOS and pregnancy status of women having ICSI cycle. This study aimed to determine the relationship between the hormone levels and pregnancy outcome in PCOS and non-PCOS women undergoing intracytoplasmic sperm injection (ICSI) program. The study was done at Higher Institute for Infertility Diagnosis and Assisted Reproductive Technologies from 2022 to 2023. Eighty infertile women participating in the IVF/ICSI treatment. They were split to control (40 non-polycystic ovarian syndrome-PCOS women) and 40 women for PCOS group. They were subdividing to subgroups’ according to body max index (BMI; normal and overweight / obese). ELISA method was used to measure hormone level at day 2 and follicular fluid and serum at day of ova pick up. About 38.8% of study participants were got pregnant. Means of serum resistin and follistatin in day 2 and OPU were significantly higher in patients with PCOS and normal BMI level while follistatin at day 2 and in follicular fluid (FF) were significantly higher in participants who got pregnant. Follistatin level at day 2 > 9.89 ng/ml and in FF > 15.06 ng/ml and might be as a predictor for occurrence of pregnancy after ICSI. The cutoff point of follistatin level in the serum at day two and in FF at the day OPU was found to be a predictor of pregnancy following ICSI, while no correlation was found between resistin and pregnancy outcome.
Amphiregulin, a growth factor inspired by LH surge, promotes oocyte maturation and follicular rupture. This study investigated whether amphiregulin growth factor levels in follicular fluid and blood serum can predict oocyte maturation and positive pregnancy rates in ICSI patients . Methods: The study included 50 women and they did ICSI from October 2024 to February 2025. Each woman got antagonist ovarian stimulation and recombinant follicle stimulating hormone. Blood samples were taken on cycle day two for infertility hormone assessment and on oocyte retrieval day for Elisa Amphiregulin growth factor estimate . Follicular fluids amphiregulin growth factor levels were significantly higher among pregnant females; however there was no significant difference of serum amphiregulin growth factor between pregnant and non- pregnant females. Out of 50 females, 11 females were had positive pregnancy test (Pregnancy rate = 22.0 %), there was a solitary positive correlation between follicular fluids amphiregulin growth factor with metaphase II oocytes count (r=0.800& p=0.003); however follicular fluids amphiregulin showed no significant correlations with other ICSI parameters. There were also no significant correlations between serum amphiregulin with all ICSI parameters. The current study found that elevated Amphiregulin (AREG) levels are linked to successful pregnancy outcomes in ICSI cycles
To explore whether dual triggering for final oocyte maturation using a low dose of recombinant human chorionic gonadotropin (hCG) plus gonadotropin-releasing hormone agonist (GnRH-a) is as effective as the standard dose of recombinant hCG triggering alone in term of a total number of retrieved oocytes, fertilization and pregnancy rate in fresh autologous antagonist intracytoplasmic sperm injection (ICSI) cycles. 80 infertile women undergoing ICSI, aged ˂ 40 years, 18˂ Body mass index ˂30 Kg/m2 included in fresh autologous antagonist ICSI cycles. 40 women received a Standard dose of recombinant hCG (500 µg) alone and 40 women received GnRH-a (0.2 mg triptorelin plus 250 µg recombinant hCG) were used for final oocytes maturation. This study was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Techniques, Al Nahrain University. The mean total number of retrieved oocytes, fertilization, and pregnancy rate were measured. The mean total oocyte number being statistically higher in the dual trigger group. The mean number of fertilized oocytes and clinical pregnancy rates was higher in the dual trigger group than in the control group, but the difference showed borderline statistical significance. Furthermore, fertilization rates show no statistical difference between the control group and the dual trigger group. Results of the study confirm that dual triggers show favorable improvement in fresh autologous antagonist ICSI cycles with the use of GnRH agonists plus hCG compared with hCG alone for triggering of final oocyte maturation.
For couples with a variety of male factor disorders, intracytoplasmic sperm injection has made pregnancy and motherhood possible. Two hundred-one males who had non-obstructive azoospermia or severe oligospermia were included in this study. One hundred forty trials of ICSI were done by using fresh sperms (fresh group n=140), while 61 trials of ICSI were done by using cryopreserved sperms (cryopreserved group n=61) to compare the conception rates between fresh and cryopreserved sperms used in intracytoplasmic sperm injection (ICSI). Twenty-five samples from each group were collected by doing a testicular biopsy. Pregnancy rates were higher using cryopreserved sperms as compared to fresh spermatozoa, 39.3% compared to 24.3% in the fresh group ( P .030), respectively. Binary logistic regression analysis showed that the odds ratio of achieving pregnancy with cryopreserved sperms was 2.022 higher than with fresh ( P .030). Cryopreservation has no negative effect on the pregnancy rate. Instead, it might improve the results of ICSI in comparison with a fresh sperm sample.
Oxidative stress in the field of reproduction has been extensively considered in association with sperm properties and male infertility; in any case, the effect of oxidative stress in association with oocyte trademark got less thought than that of sperms. The goal of the study is to understand the possible association among serum and follicular oxidative markers in infertile women and oocyte characteristics and pregnancy result in ICSI cycles. The present case-control study included 57 infertile women undergoing ICSI that was done in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies at Al-Nahrain University. The assessment started in April 2018 and connected with March 2019. Mean metaphase II oocyte count of all women was 5.70±3.87, that of pregnant women was 6.90±3.68 and that of non-pregnant women was 4.46±3.73; there was a basic qualification in mean metaphase II oocyte check among pregnant and non-pregnant women (P = 0.016); being higher in pregnant women. The mean abnormal oocyte count of all women was 0.63±1.26, that of pregnant women was 0.28±0.45 and that of non-pregnant women was 1.00±1.68. Oxidative stress markers inside and out affected clinical pregnancy result (P < 0.05) and appear to impact clinical pregnancy bring about women undergoing ICSI due to its effect of oocyte characteristics.
The vaginal microbial makeup has a substantial impact on women's health. The typical vaginal microbial profile in healthy women of reproductive age comprises aerobic, facultatively anaerobic, as well as obligately anaerobic organisms. However, the optimal vaginal microbiota is dominated by lactobacilli and is thought to defend against pathogenic microbial colonization and infection by producing lactic acid, antimicrobial metabolites, and lowering immune system activation. Recent research has come up with the term "molecular bacterial vaginosis (BV)" to describe non-optimal vaginal microbiota that is low in Lactobacillus species (spp.) and has a lot of facultative and absolute anaerobic bacteria specifically Gardnerella vaginalis. In the context of IVF, BV has been linked to infertility. The study comprised forty-six women to explore the vaginal and follicular microbiota of asymptomatic infertile Iraqi women starting an ICSI trial. At the time of ova pick-up, a high vaginal swab and follicular fluid were acquired from each instance for DNA extraction to test for the presence of microorganisms by the use of a real-time PCR special commercial kit. In conclusion, molecular technologies have offered a more thorough view into the vaginal microbiota, which may explain why vaginal dysbiosis shifts during the ICSI-ET treatment. Even though the majority of women have no clinically visible infection, the fact is that they have an aberrant vaginal micro-ecology profile. Nevertheless, isolating microorganisms from follicular fluid did not reduce the likelihood of fertilization or the pregnancy rate during ICSI cycles.
The aim of any given ICSI cycle is to achieve normal response and avoid poor or hyper response. A part from ovarian hyper stimulation syndrome (OHSS), pregnancy rate in hyper responder females usually low which could be related to the adverse effect of hyper estrogenic state on oocytes, embryos quality and uterine endometrium. To evaluate the effect of hyper response on ICSI outcome represented by oocytes, embryos quality, fertilization and pregnancy rates. Study showed that total number of retrieved oocytes, mature, immature oocytes, total embryos, good quality embryos and blastocyst count were significantly higher in hyper followed by normal and poor responders respectively. Fertilization rate was significantly higher in both hyper and normal responders than poor ones, while pregnancy rate was significantly higher in hyper responders 47.6% followed by normal 40.7% and then poor ones 16.7%. Conclusions: Irrespective of abnormal hormonal milieu, excessive ovarian response and higher number of produced oocytes improve ICSI outcome in hyper responder females by increasing the chance of getting higher number of embryos, good quality embryos and even blastocyst numbers which in turn increase the chance of pregnancy.
Stem cell factor (SCF) is one of the first growth factors derived from granulosa cells in the ovarian follicle plays a critical role in hematopoiesis and the generation of melanocytes and germ cells. SCF also serves an important role in the development of the interstitial cells of Cajal in the intestine and the learning functions in the hippocampal region of the brain. This study aimed to investigate the levels of SCF in follicular fluid (FF) and if it can be used as a potential marker for predicting oocyte, embryo quality, and pregnancy rate. A total of 44 infertile couples involved in the study. All of them underwent controlled ovarian hyperstimulation for the ICSI cycle. Antagonist protocol was used as an ovulation induction protocol. Patients were divided into two groups according to ICSI outcomes as pregnant (23 women) and non-pregnant (21 women) groups. A comparison was done in both groups based on the oocyte, embryo quality, and other ICSI cycle characteristics, and follicular fluid stem cell factor level. SCF level was measured by using an enzyme-linked immunosorbent assay. Our study showed lack of association between patients' characteristics which included (age, BMI, infertility duration, type of infertility, and causes of infertility, basal hormonal, stimulation characteristics levels), FF SCF level and the oocyte characteristics comparison (total number retrieved oocyte, germinal vesicle oocyte (GV), MI oocyte and MII oocytes, and the number of injected oocytes, fertilization rate, and embryo characteristics) between pregnant and non-pregnant groups. The present study showed that Mean follicular fluid SCF was significantly lower in pregnant women in comparison with that of non-pregnant women, 135.92±38.96 pg/ml versus 185.15±63.58 pg/ml, respectively (P=0.003). Raised SCF level in FF harms ICSI outcome.
Background: Azoospermia is defined as the lack of sperm in male ejaculate and is suggested to be present in about 1 % of all males and to account for 10-15% of the causes of infertility in men. The development of intracytoplasmic sperm injection (ICSI) as an effective therapy. Embryo grading has gained special attention in published literature regarding its predictive role in pregnancy outcome following ICSI; however, substantial controversy is present that justified the carrying out of the present study. Objective: To study the association between embryo quality and pregnancy rate in women of males with azoospermia. Patients,Materials and Methods: This prospective cohort study included 60 patients with azoospermia, 15 male with obstructive type and 45 males with non-obstructive type.Those patients were randomly selected form the population of infertile couples who regularly visits the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies and from Kamal Al-Samarrai Hospital -Center for Infertility Treatment and IVF. Results Pregnancy was encountered in 7 patients (23.33%). It was not statistically associated with type of azoospermia (P=0.388), also it was not affected by age and serum hormonal level, but highly determined by good quality of embryo. Quality of embryo was graded from 1 to 4. The lower the grade, the better the quality of the embryo was. Quality of embryo was not significantly correlated with type of azoospermia, age and hormonal levels. Conclusions Good quality of embryo is a significant predictor of positive pregnancy outcome in ICSI and the quality of embryo is not affected by age of male patient or type of azoospermia.
Plasminogen Activator Inhibitor-1 expression in serum and follicular fluid plays a crucial role in regulating fibrinolysis during the pregnancy period by blocking the action of other proteins called plasminogen activators. The aim is to assess the Plasminogen Activator Inhibitor-1 level in blood serum and follicular fluid as a predictive marker of pregnancy. This study included Sixty-nine infertile females undergone ovarian stimulation with antagonist protocol and received recombinant follicle-stimulating hormone, at the cycle day two. The serum and follicular fluid were collected from each female on the day of oocyte retrieval, for PAI estimation using the Elisa kit. The data were analyzed using version 23.0 SPSS and Microsoft Office 2010. The serum PAI-1 levels at the day of oocyte retrievals were significantly lower in positive pregnancy women in comparison with non-pregnant women (p < 0.05), Yet no significant follicular fluid PAI-1 levels were noted in comparison with non-pregnant women. There was a negative significant correlation (p > 0.05) between serum and follicular fluid PAI-1 levels on the day of oocyte retrieval according to oocyte ICSI outcome. The evaluation of serum PAI-1 levels can be used as a promising parameter for the prediction of pregnancy outcomes of ICSI patients. It is assumed that the pregnancy rate was affected by PAI-1 level during ICSI cycles. The concentration of PAI-1 in serum and follicular fluid at the retrieval day is not correlated with the oocyte's maturity.
Endometriosis is a chronic inflammatory gynecological disease and a leading cause of female infertility, characterized by Th1/Th2 cytokine imbalance. Interleukin-13 (IL-13) and Interferon-gamma (IFN-gamma) are cytokines mediating anti-inflammatory and pro- inflammatory pathways, respectively, and were investigated in this study in infertile women with endometriosis undergoing Intracytoplasmic Sperm Injection (ICSI).To explore the potential impact and prognostic value of Interleukin-13 (IL-13) and Interferon- gamma (IFN-gamma) on ICSI outcome and to confirm their role in endometriosis-related infertility .A prospective case-control study was conducted involving 80 infertile women aged 18-40 years, including (n=40) women laparoscopically confirmed with endometriosis and (n=40) infertile women without endometriosis serving as controls. Both groups enrolled in ICSI cycles, and on the day of oocyte retrieval, serum and follicular fluid were collected; then concentrations of IFN-γ and IL-13 were quantified using enzyme-linked immunosorbent assay (ELISA). Ovarian stimulation characteristics, oocyte maturity, embryo quality, maturation rate, fertilization rate, and pregnancy rate were compared between groups using appropriate statistical analyses to investigate their association with endometriosis and their impact on ICSI outcomes. Elevated IFN-γ and IL-13 levels in both serum and follicular fluid are strongly associated with impaired ovarian response and poor ICSI outcomes in women with endometriosis. These findings suggest that inflammatory cytokine imbalance contributes to reduced oocyte developmental competence and embryo quality. Assessment of IFN-γ and IL-13 may provide clinically valuable biomarkers for predicting ART outcomes and could facilitate individualized therapeutic strategies targeting immune-mediated reproductive dysfunction.
EMMPRIN (Basigin) is a potent inducer of matrix metalloproteinases (MMPs). Expression of EMMPRIN in the uterus has a very important role in implantation of embryo, this study was conducted to evaluate the relationship of pregnancy rates with EMMPRIN levels in follicular fluid and serum of intracytoplasmic sperm injection patients. The study was cross sectional study analyzed the serum and follicular fluid samples of 58 patients undergo ICSI procedure. The serum and follicular fluid samples were collected from infertile women aged 20 to 42 years. However, the levels of EMMPRIN were measured in the serum and follicular fluid samples and these samples were obtained during oocytes retrieval and another serum samples were collected at the day of embryo transfer. All women have ovarian stimulation with recombinant follicle stimulating hormone (FSH) at the second or third day of the cycle. Result: The pregnancy rate was 27.6 %. The present study showed that EMMPRIN in serum and follicular fluid at day of ova pickup and its level at embryo transfer day, are significantly low in pregnant women (p < 0.05) compared to non-pregnant women. The statistical analysis showed no significant correlation between serum and follicular fluid EMMPRIN with endometrial thickness (p > 0.05). EMMPRIN level can be used as a promising parameter for the prediction of pregnancy rate in ICSI cycles.
The 5,10-methylenetetrahydrofolate reductase (MTHFR) enzyme is a key methionine and folate metabolism enzyme that is found in oocytes and embryos in preimplantation stages and plays a vital role in female reproduction.To assess the impact of MTHFR genetic polymorphisms (C677T and A1298C) on follicular stimulating hormone (FSH), luteinizing hormone(LH), Estradiol (E2), progesterone and antimullerian hormone (AMH) in female receiving intracytoplasmic sperm injection (ICSI), 85 infertile women undergoing ICSI treatment at the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies in Baghdad, Iraq, were recruited. These patients had their MTHFR polymorphism genotypes examined, FSH, LH, E2, progesterone, and AMH levels were compared between groups. Furthermore, all demographic characteristics as well as ICSI outcomes were compared between pregnant and non-pregnant women. The research revealed no discernible variation between the means of FSH, LH, E2 and AMH with respect to the 219A>C(rs1801131) (A1298C) genotypes (p > 0.05). Progesterone level was significantly differed with high level found in CC followed by AA and AC. There was no statistically significant variation in hormonal features between CC, CT, and TT for A1298C, 219A>C(rs1801133). The demographic features (age, duration, type of infertility, cause of infertility and previous trial) were not notably different between women who were pregnant and those who were not. In women undergoing ICSI, the maternal MTHFR polymorphisms is not related to FSH, LH, E2 and AMH. MTHFR polymorphisms therefore don't offer useful information regarding the hormonal levels in our groups.
Background : The overall success rate of assisted reproductive techniques is only about 30% due to certain limiting factors. Endometrial scratching could induce inflammatory response which manifested by the up-regulation of different cyto kines and chemokines with subsequent accumulation of immune cells in the in jured site. One of the most important cytokine in this regard is TNF-α which was found to induce tissue remodeling by stimulating the production of many other pro-inflammatory cytokines. Objectives: To investigate the role of endometrial injury during the luteal phase before ICSI cycles in the enhancement of pregnancy outcomes. Materials and Methods: This prospective study was undertaken in the High In stitute of Infertility diagnosis and Assisted Reproductive Techniques / Al-Nahrain University. All women were undergone ICSI cycles. Women in the intervension group underwent endometrial scratching with a Pipelle catheter in luteal phase preceding cycle. Blood sample was taken before scratch. All patients undergo controlled ovarian hyperstimulation, oocyte retrieval and embryo transfer, anoth er blood sample was taken at day of embryo transfer to measure level of TNF-α. Clinical pregnancy rate then evaluation after luteal support of two weeks. Results: The patients in scratch group has higher pregnancy rate (14/23) 60.9% than non intervention group (11/33) 33.3% although not significant (p value 0.057). The results showed that the TNF-α after scratch (54.53±10.2) was significant ly higher than before scratch (39.26±14.74) in the same group, as p value was ( <0.001). In addition the TNF-α was significantly higher in scratch group (54.53±10.2) than control (25.91±9.08) as p value was (< 0.001). Conclusion: Induced endometrial local injury is an easy, simple and cost-effec tive technique that can be used to improve the uterine receptivity and enhance embryo transplantation.
The luteal phase (LP) in the fresh ICSI cycle is insufficient, adequate LP support is one of the approved treatments for improving implantation and pregnancy rates. It is generally known that the LP is inadequate after ovarian stimulation due to negative from supra-physiological blood levels of steroids released by numerous corporal luteal, LH concentrations are low during the luteal phase. In this study, patients were divided into two groups: (40) patients as study group; those who received GnRHa (Decapeptil 0.1 mg), three days after embryo transfer, in addition to conventional luteal phase support (LPS) in the LP to increase the implantation and pregnancy rate in IVF; and their control group (40) received standard LPS only. On the second day of stimulation, blood samples for FSH, LH, TSH, E 2 , and prolactin were taken. On the day of ovulation induction, measure E 2 , progesterone, and LH; and on the day of embryo transfers, measure progesterone and LH. The overall characteristics of the patients in both groups were not significantly different. There was also no significant change in the number of total oocytes, mean of metaphase II oocytes percent, cleavage rate, grade I embryo percent, or serum hormones level between the study and control groups (p > 0.05). GnRH agonist treatment in the luteal phase improves clinical pregnancy and implantation rate in fresh ICSI cycles but is not statistically significant.
Receptive endometrium is essential for successful embryo implantation and is created by the synergistic activities of estrogen and progesterone, as well as support from other endocrine, paracrine, and autocrine pathways. Granulocyte Colony Stimulating Factor (G-CSF) may play a crucial role in modulating the immune response to enhance implantation. To estimate the role of granulocyte-colony stimulating factor on the clinical intracytoplasmic sperm injection (ICSI) outcome parameters. A prospective comparative study with random sample selection was carried out from 2021 to 2023 at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. It included 80 women with infertility, with or without previous failure trials of intracytoplasmic sperm injection, who completed IVF and ICSI protocols and reached the day of embryo transfer. On the day of embryo transfer, patients were divided into two groups: 49 patients received no additional treatment (group 1), and 31 patients received granulocyte-colony stimulating factor subcutaneous injection within one hour of embryo transfer (group 2). The implantation rate was 13.3% in group 2 compared to 7.6% in group 1. The chemical and clinical pregnancy rates were 29.0% and 25.8% in group 2, while group 1 had 18.4% for both chemical and clinical pregnancy rates. The pregnancy and implantation rates were slightly higher in group 2, but the difference was not significant. More research is needed to support the use of G-CSF as a standard treatment in ICSI patients.
Infertility is the disability of a couple to obtain pregnancy after one year of sexual intercourse that is unprotected and regular. Bisphenol A is an industrial chemical spread worldwide, that is a raw material utilized in multiple industries. Bisphenol A is assorted as an endocrine-disrupting chemical, which can interfere with hormone activity. This study aimed to assess the effect of BPA levels in serum on ICSI outcomes. The study was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University during the period from October 2021 to April 2022. sixty infertile women enrolled in this study. BPA levels were analyzed through an enzyme-linked immunosorbent assay (ELISA) technique. Mean serum BPA was not significantly higher in non-pregnant women in comparison with that of pregnant women. Serum BPA was positively correlated to immature oocytes (MI) and abnormal oocytes. Also, BPA inversely correlated to Grade1 embryos and positively correlated to Grade 3 embryos. Bisphenol A can be detected in sera of infertile female undergoing ICSI. High BPA level in serum may impact oocyte quality and embryo grading
Background:Defects in reproduction that may involve both the male & female partners result in the inability to procreate which is known as infertility. The use of assisted reproductive technologies (ART’s) has been increasing over the past three decades. Many efforts have been done to compensate for unsatisfac-tory results in in-vitro fertilization (IVF) procedures, so that patients undergo different ovarian stimulation protocols to control retrieval of multiple oocytes in a single cycle. Although ovarian stimulation has an important role in ART’s, it may also have detrimental effects on oocyte quality, embryo quality, and IVF outcomes. Objectives: To assess the effect of long agonist versus antagonist protocols on oocyte and embryo quality in IVF-Intracytoplasmic sperm injection (ICSI). Patients,Materials and Methods: This comparative study was done on the embryos of seventy one infertile fe-males who were undergoing Intracytoplasmic sperm injection (ICSI) treatment regardless to the presence or absence of previous trials. These patients were divided into two groups, 45 of them treated with agonist protocol and other 26 patients treated with antagonist protocol. First oocyte retrieved was used for further assessment. Of the 71 oocytes, there were eight immature oocytes (3 at germinal vesicle (GV) stage, 5 at metaphase I (MI) stage) and three abnormal oocytes, and these were excluded from the following ICSI treatment. Assessment and comparison of oocyte maturity, fertilization rate, cleavage rate and resulting embryo quality were done. Results:The results of the present study showed significant difference (p<0.05) in oocyte number and maturity but no significant (p>0.05) difference between the group treated with long agonist and those on antagonist protocol regarding fertilization rate, cleavage rate, and embryo quality. Conclusions:The type of protocol used in in-vitro fertilization has no detrimen-tal effect on oocyte or embryo quality.
Reactive oxygen species (ROS) are believed to play an important role in the function of the male and female reproductive system, as well as in embryonic development. If the balance between production and elimination of ROS is disturbed, pathogenic consequences would happen. Reactive oxygen species can sometimes exert damaging effects when overabundant, oxidative stress is hypothesized to have a negative impact on, for example, folliculogenesis, embryo development, oocyte maturation, oocyte fertilization, endometriosis, polycystic ovary syndrome and unexplained infertility .8-Hydroxy2-desoxy guanosine (8‐OHdG) is a sensitive indicator of DNA damage as a result of oxidative stress. The study aim was to evaluate follicular fluid levels of 8-Hydroxy2-desoxy guanosine in patients undergoing Intracytoplasmic Sperm Injection (ICSI) and compared the results between both pregnant and non-pregnant patients . One hundred and twenty-seven women included in this study, they underwent controlled ovarian hyper stimulation, program followed by ova pick up, at that time follicular fluid were obtained and freezes till the time of analysis for reactive oxygen species (ROS), patient injected oocyte was cultured in CO2 incubator, then embryo grading, embryo transfer, pregnancy rate and reactive oxygen species level were determined. Follicular fluid 8-OHdG was significantly lower in pregnant females (p < 0.001), 8-OHdG had negative correlations with grade 1 and grade 2 embryos and negative correlation with total blastocyst count. Follicular fluid levels of 8-OHdG provides sensitive tool to predict positive pregnancy outcome in women undergoing ICSI.
Cigarette smoke contains a substance which is a toxic mutagenic chemical, and carcinogenic compounds, which have a negative effect on male fertility. Smoking cigarettes is associated with noticeably higher levels of the oxidative stress-causing reactive oxygen species in the seminal fluid. Because of this, smoking may cause changes to the sperm plasma membrane and a significant amount of deoxyribonucleic acid fragmentation. To study the influence of smoking on sperm chromatin immaturity and ICSI outcomes (fertilization rate, embryo grading and pregnancy rate). At the laboratories of the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies and after 1-7 days of sexual abstinence, 92 males’ sperm samples were taken. After liquefaction, macro-and microscopic studies were carried out, criteria applied by the WHO 2021. Masturbation was used to obtain the sample, which was then placed in sterile containers. The relationship between the smoking and chromatin immaturity and ICSI outcomes have been evaluated. A significant positive correlation between smoking status and sperm chromatin immaturity. Furthermore, there was a negative non-significant correlation between smoking status and fertilization rate. Weak and non-significant correlations between smoking status and embryo grade 2 as well as embryo grade 3. There was a positive association between smoking status and embryo grade 1. The sperm chromatin integrity was significantly lower in the non-smoker group compared to the smoker group.
The estimation of oocyte quality in human in vitro fertilization (IVF) is an important point for the embryologists. Oocyte selection and the identification of the best oocytes may help to limit an overproduction of embryos and to improve the results of oocyte cryopreservation programs. The follicular fluid (FF) can be provided easily during oocyte pick-up and known to represents an optimal source for non-invasive biochemical predictors of oocyte quality. However, till now no substance was found to be used as reliable markers of oocyte competence to fertilization, embryo development and pregnancy. Metabolism and ATP levels within the oocyte and adjacent cumulus cells are associated with quality of oocyte and optimal development of a healthy embryo. Lipid metabolism provides a potent source of energy and its importance during oocyte maturation is being increasingly recognised. Objectives: To determine the effect of lipids (Cholesterol. Triglyceride and HDL) level in serum and follicular fluid in predicting oocyte quality, embryo quality and outcome in patients undergoing intracytoplasmic sperm injection program Patients and Methods: Fifty eight (58) infertile women with an age range 21-41 years old undergoing intracytoplasmic sperm injection cycles were included in this study. Lipids (Cholesterol. TG and HDL.,) levels in serum and follicular fluid were measured by auto analyzer for biochemistry (Flexor El_200) and correlation with oocyte quality, embryo quality and ICSI outcome was done. Results: The results of this study showed no significant (p<0.05) difference in lipid levels in serum and follicular fluid between pregnant and non pregnant females as well as non significant correlation was found between lipid level in serum and follicular fluid in relation to oocyte quality and embryo quality. Conclusions: Serum and follicular fluid lipids are not good predictors of oocyte quality, embryo quality or ICSI outcome in women undergoing IVF/ICSI cycles. Accordingly, these biomarkers would be less reflective of the follicular environment and that the effect of maternal metabolic disorders on oocyte quality and fertility outcome is complex and of a multifactorial kind.
Due to malfunction of the hypothalamus-pituitary-ovary axis and probable dysregulation of ovarian activities, ovary has a large number of small follicles but no developed follicles. L-Carnitine is a tiny, water-soluble chemical that plays a critical role in fat metabolism. It is required for appropriate mitochondrial fatty acid oxidation and acyl-CoA ester excretion, as well as affecting ATP levels. The current study was aiming at evaluating the protective effect of L-Carnitine (LC) against deleterious substances present in serum of patients with PCOS, which may affect the oocyte maturation. 60 infertile women diagnosed with polycystic ovarian syndrome have been enrolled in this study and enter their Intracytoplasmic Sperm Injection (ICSI) cycle; they were informed about the study and signed a written informed consent. They were grouped into treatment group (n=30) who were treated with L-Carnitine® tablet for two months and placebo group (n=30). All enrolled women underwent flexible antagonist protocol for ICSI. Significant increase in mean mature metaphase II (MII) oocyte count in study group than placebo group, oocyte maturity rate was significantly higher in study group in comparison with placebo group. In addition, mean fertilized (2PN) oocyte count was increased significantly in study group in comparison with placebo group. Mean fertilization rate was significantly higher in study group in comparison with placebo group. Treatment with of patients with polycystic ovary syndrome of L-Carnitine resulted better oocyte quality and the improvement in oocyte quality is reflected by mature MII oocytes, maturity rate and fertilized rate with eventual improvement in embryo quality.
Non-steroidal anti-inflammatory medications may interfere with ovulation, implantation, and pregnancy; they are generally avoided during reproductive procedures. The risks of using them during IVF must be carefully weighed against the necessity of pain relief during oocyte retrieval. About 64 women (ages 20–42) undergoing ICSI participated in a single-blind, cross- sectional clinical trial at the High Institute for Infertility at Al-Nahrain University from October 2024 to May 2025. During oocyte retrieval, participants were split into two groups at random and given propofol anesthesia and ketorolac analgesia, respectively. They all adhered to a common protocol for ovarian stimulation. Based on their maturity stage (MII, MI, or GV), oocytes were categorized; those that were aberrant or degenerated were not included in ICSI. ELISA was used to measure the levels of cortisol and serum LH both before and after oocyte retrieval. The mean post -operation LH Marker level were 10.41±8.0, 6.40± 7.48 among GA and analgesia groups respectively, this relation was statistically significant (P value =0.04). The mean Post -operation Cortisol Marker level were 201.13±132.34, 159.13± 78.42 among GA and analgesia groups respectively . The mean number of Oocytes: Similar for both groups (10±5.53 for anesthesia and 10.59±5.44 for analgesia). Abnormalities of Oocytes: slightly lower in the group receiving anesthesia (0.56±1.44 vs. 0.69±1.18). The anesthesia group had fewer ruptured oocytes (0.16±0.45 vs. 0.25±0.57). Fertilized embryos: Grade I: The anesthesia group had slightly fewer (2.75±2.63 vs. 3.13±2.46). Grade II: Groups are comparable (1.41±1.58 vs. 1.31±1.53). Grade III: Lower in the group under anesthesia (0.19±0.54 vs. 0.38±0.83). Similar results were obtained from arrested embryos (anesthesia: 0.63±1.1, analgesia: 0.68±1.64). The analgesia group had a significantly higher number of transferred embryos (anesthesia: 1.84±1.08, analgesia: 2.88±1.62; P = 0.004). The analgesia group showed a significant decrease in post-operation LH marker, Cortisol Marker, and higher mean number of transferred embryos compared to the anesthesia group.
Background : With the rapid evolution of assisted reproductive technologies, more and more studies are concerned about the possible influential factors on the success rates of these procedures, with the efforts are now mainly directed toward defining these factors in a trial to increase these rates. Of these factors, inflammation is considered among the most remarkable and is in the focus of the updated studies. However, most of these studies are evaluating the role of inflammation in patients with polycystic ovarian syndrome(PCOS) , while the impact of this inflammation in insulin resistant non PCOS patients is underesti mated and not well evaluated . Objectives: This study is a trial to elucidate the importance of one of the inflam matory mediators , Tumor Necrosis Factor-alpha (TNF – α) , in insulin resistant patients undergoing IVF ( ICSI ) cycles . Patients and Methods: A group of 27non-PCOS infertile patients were select ed for this clinical randomized prospective study from patients attending the high institute for infertility diagnosis and assisted reproductive technologies / Al-Nahrain University and subjected to intra-cytoplasmic sperm injection (ICSI) procedure. BMI was measured for all patients and they were classified according to WHO classification into 3 groups : Acceptable , overweight and obese . Fast ing plasma level of glucose and insulin was investigated and insulin resistance index (HOMA) was calculated. Serum TNF-α has been measured using ELISA and the relation with insulin resistance was investigated. Finally, the pregnancy rate, as a primary reproductive endpoint was compared according to TNF -a and insulin resistance levels between different study subjects. Results: Of the 27 infertile patients , 17(63 %) were non obese and 10 (37%) were obese , 15(55.5%) were having HOMA index < 1.6 and 12 (44.5 %) were having HOMA index ≥ 1.6 . BMI was found to be positively correlated with high er HOMA index ( P = 0.01 and R= 0.4 in spearman correlation). Investigating the relation between TNF-α with BMI revealed that TNF-α levels are increasing with the increment in BMI, However , the difference in the mean TNF-α between the three BMI groups failed to achieve statistical significance (P=0.1). In contrast to that, when stratifying for HOMA index , TNF-α was significantly correlated with it ( spearman correlation R= 0.4 , P=0.01 ) , and patients with HOMA index ≥1.6 were found to have higher levels of TNF-α ( P = 0.03) and to be more obese ( P=0.02 ) than those with HOMA index <1.6.
Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility and is associated with an increased risk of premature luteinizing hormone (LH) surges during controlled ovarian stimulation. Progestin-primed ovarian stimulation (PPOS) using Medroxyprogesterone Acetate (MPA) has emerged as a convenient alternative to conventional gonadotropin-releasing hormone (GnRH) antagonist protocols. This study aimed to compare the effects of MPA-based PPOS and GnRH antagonist protocols on oocyte yield, maturation, fertilization, and embryo quality in women with PCOS undergoing ICSI . This prospective, randomized, open-label controlled clinical trial included 108 women with PCOS undergoing ICSI at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, Baghdad, Iraq, between 2023 and 2024. Participants were allocated to an MPA-based PPOS group (n=49) or a flexible GnRH antagonist group (n=59). Oocyte yield, metaphase II (MII) oocytes, fertilization rate, embryo quality, ovarian sensitivity index (OSI), gonadotropin consumption, stimulation duration, and LH profiles were evaluated.No significant differences were observed in baseline characteristics or hormonal profiles between groups. The MPA group showed a numerically higher number of retrieved oocytes (20.65 ± 7.50 vs. 13.39 ± 4.41; p=0.083) and MII oocytes (15.10 ± 5.20 vs. 11.24 ± 4.12; p=0.09), although these differences were not statistically significant. Fertilization rates were comparable between groups (72.4 ± 10.6% vs. 71.2 ± 11.3%; p=0.68), as were embryo quality, OSI, gonadotropin consumption, and stimulation duration. Both protocols effectively suppressed premature LH surges without premature ovulation or cycle cancellation. MPA-based PPOS appears to be an effective and practical alternative to GnRH antagonist protocols for PCOS patients undergoing ICSI, providing comparable fertilization and embryological outcomes with effective LH suppression. Its oral administration and potential cost advantages may be particularly valuable in resource- limited ART settings.
The selection of protocols in controlled ovarian stimulation during intracytoplasmic sperm injection affect the treatment outcomes. Both GnRH agonist and GnRH antagonist protocols are clinically utilized, therefore comparision of these protocols is important for the evaluation of their laboratory and clinical outcomes.To compares the short GnRH agonist and GnRH antagonist protocols effectiveness in expected normal responder females undergoing intracytoplasmic sperm injection (ICSI). Methods: A retrospective study was involved 60 women aged less than 35 with antimullerian hormone (AMH) more than 2.5 ng/ml. Participants women were assigned to either group1 short GnRH agonist protocol and group2 GnRH antagonist protocol. The outcomes measured including total oocyte, mature and immature oocytes, number and quality of embryo and pregnancy rate.The short GnRH agonist protocol group showed statistical significant benefit in oocyte yield (p- value 0.04), metaphase I (p- value 0.09), metaphase II (p- value 0.03), embryo number (p- value 0.04), embryo quality (p-value 0.01) and pregnancy rate in expected normal responder women compared to GnRHantagonist protocol. The short GnRHagonist protocol reveals favorable ICSI outcomes than antagonist protocol and may be better choice in expected normal responder women.
Granulosa cells produce most ovarian function indicators in infertile women like estradiol, AMH, and inhibin B. Female peptide hormone insulin-like factor 3 is produced by theca cell. Signaling of insulin-like factor 3 from theca cell stimulates antral follicle development and theca androgen production. To investigate how INSL3 levels may be a theca cell marker of ovarian reserve and the association with current markers of ovarian reserve, including AFC, FSH, E2, and AMH, in infertile women undertaking assisted reproduction. In this cross-sectional study, 75 Iraqi infertile couples were chosen at Al-Nahrain University's High Institute for Infertility Diagnosis and Assisted Reproductive Technologies from October 2022 to April 2023 for the ICSI cycle. Based on the levels of INSL3 in the serum on the day 2 of the menstrual cycle and on the day of retrieving oocyte in the follicular fluid. A comparison was made with current ovarian reserve markers (AFC, FSH, E2, and AMH) .From 75 infertile women undergoing intracytoplasmic sperm injection (ICSI), 21 of them achieved pregnancy. Positive Significant relations were found between serum INSL3 levels and anti-mullerian hormone (AMH and antral follicle count. there were no significant relationships between follicular fluids INSL3 with serum LH, FSH and E2 . The study provides compelling evidence for the significant role of INSL3 in female fertility and its association with the success of intracytoplasmic sperm injection (ICSI). Positive correlations between serum INSL3 levels and anti-mullerian hormone (AMH) and Antral follicle count AFC indicate its potential involvement in ovarian reserve and hormonal regulation.
Glycotoxins termed as advanced glycation endproducts (AGEs) are made endogenously and exogenously. Maillard reaction is a chemical non-enzymatic process, between sugars and a free amino acid group of nucleic acids, proteins, and lipids which happens exogenously when handled with high temperature for a prolonged period and in a dry environment, resulting in glycotoxins (AGEs) formations. This reaction is responsible for the taste, color, and smell of food. Glycotoxins derived from food are absorbed in the intestine and store in cells and tissues. Also, glycotoxins are derived exogenously from smoking. While endogenous glycotoxins are generated by normal body physiology. Obesity and insulin resistance may induce glycotoxins (AGEs) accumulation. The objective of this study was to evaluate the relationship between glycotoxins (AGEs) and obesity. 66 infertile females who were undergoing intracytoplasmic sperm injection ICSI. These females were classified into three groups according to their body mass index. On the day of ova pick up the collect serum and follicular fluid for subsequent measurement of glycotoxins (AGEs) by ELISA kits. there was a significant difference in the concentration of glycotoxins in follicular fluid and serum between the three ICSI groups of patients. As well as there was a significant difference in the concentration of glycotoxins (AGEs) in follicular fluid and serum in the same ICSI groups of patients. there is a significant positive relation between glycotoxins (AGEs) and increase BMI.
During the process of embryo development, endometrial receptivity is very important for the successful implantation of blastocysts. This research investigated the influence of endometrial thickness on the day of oocyte retrieval and fresh embryo transfer on pregnancy outcomes in Iraqi women who are undergoing intracytoplasmic sperm injection (ICSI). Women from Iraq are taking part in this investigation. Participating in in vitro fertilization (IVF) cycles with the use of ICSI was a group of 44 women who were diagnosed with polycystic ovarian syndrome (PCOS) and had unexplained infertility. During the phases of oocyte retrieval and embryo transfer, there were significant differences seen between the two groups (p-values of 0.003 and 0.049), respectively. There was a total of 44 individuals, and 31 of them had negative pregnancy test results. The remaining 13 persons had positive findings. These results illustrate the importance of endometrial thickness in influencing the effectiveness of assisted reproductive technologies in achieving pregnancies.
Serum chemerin levels are associated with body fat percentage and BMI, and elevated chemerin levels are associated with metabolic problems and inflammatory shifts. This study aimed to evaluate the effect of BMI in serum and follicular fluid chemerin and Leptin levels on patients undergoing ICSI . This study was performed by 44 women who underwent ICSI procedures and divided into two groups based on BMI: the non-obese group, with a BMI<30 and the obese group, with a BMI>30. The result in the non-obese group showed a significant positive correlation was found between Serum chemerin levels, indicating a strong effect size (p< .001). This suggests that as Serum chemerin levels rise, so will FF chemerin. A significant positive correlation of r =0.92 was found between the oocytes retrieved and the Number of metaphases II, indicating a strong effect size (p<.001, This shows that the number of metaphase II oocytes increases as the number of retrieved oocytes increases. A significant positive correlation of 0.66 was found between the number of embryos and the G1 embryo, indicating a large effect size. Conclusion: Serum chemerin had a significant correlation with oocyte maturity and embryo grading, and there was no significant difference between obese and non-obese patients regarding serum chemerin pregnancy.
Human chorionic gonadotropin is a glycoprotein hormone produced by the placenta. It exists in several isoforms with long half-lives (hours) and angiogenic potential; it maintains the corpus luteum to produce progesterone. This study aimed to evaluate Human chorionic gonadotropin concentration in follicular fluid and its effect on oocyte maturity and pregnancy rate, and whether it can be used as a noninvasive biomarker for predicting oocyte competence and pregnancy rate during intracytoplasmic sperm injection cycles. Infertile women undergoing ICSI treatment were included in the study. An antagonist protocol was used for ovulation induction. Patients were divided into two groups: pregnant women (n=18) and non-pregnant women (n=22). Samples of follicular fluid were collected on the day of oocyte pick-up. Chemiluminescent Immunoassay (CLIA) equipment was used for the quantitative determination of total β-human chorionic gonadotropin in follicular fluid. The mean beta-human chorionic gonadotropin level showed a highly significant increase relative to the mean level of non-pregnant women (110.24 ±20.50) versus (56.195 ±29.45). Moreover, there was no significant relationship between the levels of beta-human chorionic gonadotropin follicular fluid and the total number of oocytes and oocyte quality in the two groups. The Beta Human chorionic gonadotropin concentration in the follicular fluid plays an important and influential role in the prediction of pregnancy rates. Despite this, follicular fluid hCG level showed no relationship with the total number and quality of oocytes in the pregnant and non-pregnant groups.
Reactive oxygen species (ROS) are active chemical compounds created by the body's metabolic process and are crucial in human reproduction. Excessive ROS can contribute to oxidative stress, which has been linked to poor fertility. Embryo culture medium plays an essential role in IVF/ICSI cycles. Eighty participants underwent ovarian stimulation and ICSI cycles; ROS levels were measured in blood samples on the ovum pickup (OPU) day and in each embryo culture dish on ET day using enzyme-linked immunosorbent assay (ELISA). These values were associated with the quality of generated embryos. Women were divided into two groups; the first had day three embryo transfers, whereas the second received blastocyst transfers on day 5. Grades I embryo count was shown to be inversely related to the serum and culture medium ROS levels. Pregnant women had significantly greater levels of GI embryos (p=0.011) and lower levels of GIII embryos (p=0.024). Pregnancy rates for the third-and fifth days of the embryo transfers were 24% and 50%, respectively, with (p=0.045). Pregnancy success rates were increased after ET day five and were strongly influenced by transfer day. Blastocyst-stage embryos with high serum ROS levels were found to be underdeveloped. Culture medium ROS levels can be used as an adjuvant criterion for embryo selection and metabolic marker for developing embryos.
Human chorionic gonadotropin is a glycoprotein hormone produced by the placenta. It exists in several isoforms with long half-lives (hours) and angiogenic potential; it maintains the corpus luteum to produce progesterone. This study aimed to evaluate Human chorionic gonadotropin concentration in follicular fluid and its effect on oocyte maturity and pregnancy rate, and whether it can be used as a noninvasive biomarker for predicting oocyte competence and pregnancy rate during intracytoplasmic sperm injection cycles. Infertile women undergoing ICSI treatment were included in the study. An antagonist protocol was used for ovulation induction. Patients were divided into two groups: pregnant women (n=18) and non-pregnant women (n=22). Samples of follicular fluid were collected on the day of oocyte pick-up. Chemiluminescent Immunoassay (CLIA) equipment was used for the quantitative determination of total β-human chorionic gonadotropin in follicular fluid. The mean beta-human chorionic gonadotropin level showed a highly significant increase relative to the mean level of non-pregnant women (110.24 ±20.50) versus (56.195 ±29.45). Moreover, there was no significant relationship between the levels of beta-human chorionic gonadotropin follicular fluid and the total number of oocytes and oocyte quality in the two groups. The Beta Human chorionic gonadotropin concentration in the follicular fluid plays an important and influential role in the prediction of pregnancy rates. Despite this, follicular fluid hCG level showed no relationship with the total number and quality of oocytes in the pregnant and non-pregnant groups.
Pregnancy cannot develop without a coordinated stability between an invading trophoblast and a receptive maternal decidua. Both the maternal decidua and the trophoblast, which is embryonic, must incessantly regulate and change one another during the processes of implantation, trophoblast invasion, and placentation. The goal of this study was to determine whether intrauterine infusion of hCG at various doses on the day of ovum pick-up would affect endometrial receptivity markers, biochemical markers, and ICSI outcome. To this end, measurements of serum levels of TNF and IL-10, endometrial thickness using ultrasound, and sub-endometrial blood flow indices using Doppler ultrasound were used.Ninety infertile women were randomly assigned to one of three groups: the research group (n = 60) was given intrauterine administration of either 700 IU or 500 IU of hCG on the day of ova pickup. The control group (n = 30) without hCG. On cycle days 2- 3, a plasma hormone level assessment was performed on each patient.In the 700 hCG group, the pregnancy rate was statistically higher (53.3% and 40%in the 500IU group respectively) as compared with the control group C (26.7%), no significant difference in ICSI outcome between group A &B. The RI and PI were significantly lower in group A (700IU) compared to group B and C. The biochemical marker TNA and ratio of TNF/IL-10 also lower in group A confirming role of Hcg in enhancement of implantation. Intrauterine injection of hCG at day of ova pickup significantly improved the endometrial receptivity marker and this effect occur in dose dependent manner
Pro-fusion proteins as mitofusins-1 are required for controlling mitochondrial shape which determined by a dynamic balance between organelle fusion and fission and also supports oocyte development. When compare with somatic cell, mitochondria of oocyte are tiny and circular in presence. The aim is to study the mitofusin-1 in the follicular fluid as a marker for evaluating of oocyte maturation in women undergoing ICSI cycles. Fifty infertile females were included in cross-section study was undergoing ICSI procedure with age 20 to 42 years. After retrieval of oocyte and the number of oocytes was recorded by embryologist and follicular fluid Samples were used for the measurement of Mitofusin 1. Mitofusin 1 levels by ELISA kit (Mybiosource /USA).The results showed considerable higher mitofusin-1levels in patients with good oocytes quality (3.71 ± 1.35 vs. 2.45 ± 1.12 & p=0.001). There were much positive correlations between follicular fluids mitofusin-1 with both total oocytes count (r= 0.374 & p= 0.007) and MII oocytes (r=0.383 & p=0.006); and there was no much correlation between mitofusin-1 levels with MI oocyte (r= - 0.100 & p= 0.490), germinal vesical oocyte (r=0.103 & p= 0.475) and fertilization rate (r= 0.054 & p= 0.711). High follicular fluid levels of Mitofusin 1 may positively impact oocyte development and pregnancy rate.
Activin is a member of the transforming growth factor-beta superfamily and is a polypeptide growth factor. Activin regulates follicle growth, gonadotropin responsiveness, steroidogenesis, oocyte maturation, ovulation, and corpus luteum function in the ovary via a local autocrine/paracrine effect. Activin measurement A level in serum and follicular fluid during the preovulatory stage in PCOS women undergoing an IVF/ICSI cycle to predict fertilization capacity earlier Fifty infertile couples, 25 with PCOS and 25 without PCOS, who attended Al Nahrain University in Baghdad, Iraq's High Institute of Infertility Diagnosis and Assisted Reproductive Technologies. The females that participated were between the ages of 18 and 40. On the day of oocyte retrieval, the level of Activin A in serum and follicular fluid was measured. Activin A levels in PCOS women's serum (activin A=1274 77.9) and follicular fluid (activin A =1038 50.3) compared to non-PCOS women's serum (activin A=453 25.74) and follicular fluid (Activin A=450 23.73). The study shows no correlation between activin A hormone with oocytes characteristics and fertilization rate in PCOS women with a significant increase of activin A hormone in the serum and FF of PCOS women compared to non PCOS women. Activin A level can predict a diagnosis of PCOS in addition to positive predictive pregnancy but not indicator of fertilization capacity in PCOS women.
The crosstalk between a receptive endometrium and a functional blastocyst during human embryo implantation is crucial for conception. Because platelets rich plasma (PRP) with concentrated platelets 4-5 times higher than normal, when release of granules containing growth factors including VEGF, TGF, PDGF, IGF, and EGF, these factors involved for sub endometrial angiogenesis and endometrial receptivity. 44 women under the age of 40 were given antagonist ovarian stimulation treatments, the oocytes were harvested utilizing 2-D power doppler ultrasound guidance, then ICSI done for them. On the hCG day all had an intrauterine autologous PRP infusion. Power Doppler is utilized to measure endometrial thickness (EnT), pulsatility index (PI), and resistance index (RI) of sub endometrial arteries, as well as serum levels of VEGF and EGF were measured on the hCG day and ovum pickup (OPU). The ultrasound findings, on the hCG day compared to those of OPU in all women with intrauterine infused autologous PRP were highly significant (HS). The mean EnT, RI, PI, VEGF and EGF at OPU day was lower than that on hCG day in a HS manner, with (p < 0.001), (p < 0.001), (p=0.047), (p < 0.001), and (p<0.001) respectively. These characteristics were shown to be significantly and independently associated to intrauterine PRP infusion. After PRP injection, both growth factors serum levels (VEGF and EGF) increased, and ultrasonography sub-endometrial metrics such as EnT, RI, and PI changed as the thickness grew while vascular resistance decreased, and all considered as predictors of endometrial receptivity.
The association between the two hormones (progesterone and Activin A) in infertile women on the day of ova pick up and their relationship with pregnancy status in assisted reproductive programs are both poorly understood. To determine the correlation of progesterone and activin A on the day of ova pick up with pregnancy status of women undergoing IVF/ICSI cycle. The study was conducted on 80 infertile couples. The serum at day of ova pick up was collected. Serum progesterone and activin A analysis were done for all samples for activin A. Out of eighty infertile females twenty-five became pregnant (Pregnancy rate = 31%). Serum progesterone levels at days of oocytes pick up was significantly higher in pregnant females than non-pregnant (9.22 ± 0.54 vs. 4.47 ± 0.44; p < 0.001). There was a significantly higher serum activin levels at day of oocytes pick up in pregnant women compared to non-pregnant (2.19 ± 0.04 vs. 1.52 ± 0.04; p < 0.001). The cut-off value of serum progesterone at day of oocytes picks up was > 7.5 ng/ml and the cut-off value of serum activin A at day of oocytes pick up was > 1.85 ng/ml. Pregnant women had higher levels of the hormone progesterone and Activin A than non-pregnant women, which is significantly correlated. This result can be utilized to predicate the pregnancy status early in IVF Centers.
Successful implantation needs a receptive endometrium, a functional and normal embryo at the stage of blastocyst developmental and a synchronized dialogue between embryonic tissues and maternal tissues. The current study was planned and conducted to investigate the potential role of vaginal pessary of hyaluronic acid on intracytoplasmic sperm injection (ICSI) outcome. This study included 63 infertile women (32 women as study group receiving hyaluronic acid and 31 control group) who were subjected to ovulation induction programs with gonadotropin releasing hormone (GnRH) antagonist protocols for in vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI) procedures. Blood samples were taken by Vacutainer tubes and centrifuged according to the manufacturer’s instructions for the preparation of serum samples. Serum samples for Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) were obtained from each patient when the basal hormonal status was determined. The second time point is when the samples were obtained on the day of ova pickup, after that the G1 embryos will be transferred inside the uterine cavity. Then performance of endometrial receptivity parameters or patterns evaluation and patients were followed up to do pregnancy test after14 days after embryo transfer. On the day of human chorionic gonadotropin (hCG), there was no significant difference in mean serum Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) between hyaluronic acid group and reference group, respectively (p = 0.824). There was no significant difference in mean follicular fluid Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) between the hyaluronic acid group and reference group, 0.92 ±0.36 pg/ml versus 0.82 ±0.22 pg/ml, respectively (p = 0.223). Treatment with the hyaluronic acid resulted in non-significant increase in serum and follicular fluid Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), and this increment is not related to Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1) improvement in local blood endometrial blood flow as reflected by reduction in resistive index and pulsatility index with eventual improvement in pregnancy outcome.
Obesity induces metabolic dysfunctions in bodily systems and is a significant health challenge. Excessive adipose tissue accumulation encompasses dynamic, endocrine, and biologically active processes. The fundamental pathogenic mechanism of obesity is chronic low-grade inflammation exacerbated by pro-inflammatory biomarkers such as free fatty acids, lipopolysaccharides, adipokines, and cytokines. The rising prevalence of obesity over the past forty years has led to an increase in associated comorbidities, including polycystic ovarian syndrome (PCOS). To clarify the impact of increasing body mass index (BMI) in PCOS and non PCOS on the ICSI outcomes of 143 infertile Iraqi females, dividing them into two main groups: PCOS and non PCOS and each group into normal weight and overweight/obese group. Group 1= normal BMI PCOS, group 2= overweight/ obese PCOS, group 3= normal BMI non PCOS, group 4= overweight/obese non PCOS. Significant differences in ICSI outcomes were observed in the PCOS groups compared to the non-PCOS groups concerning the total number of harvested, MI, MII oocytes, GV, abnormal and fertilised oocytes, total and D3 grade I embryos alongside higher number of arrested and transplanted embryos. There are numerically higher number of D3 grade II and III embryos, morula and blastocyst with no significant difference seen. The rates of maturation and fertilisation exhibited no significant disparities. The biochemical pregnancy rate was higher in group 2 compared to group 1, and in group 4 compared to group 3, with a statistically significant difference. A higher BMI among younger PCOS individuals may be associated with better pregnancy outcomes.
The compromise of uterine receptivity is one of the main reasons for the failure of assisted reproduction techniques. Leukemia inhibitory factor is one of those molecular parameters that allow us to identify an endometrium as receptive. Platelet rich plasma (PRP) contains a range of cytokines and growth factors including leukemia inhibitory factor that encourage the migration, proliferation, and differentiation of cells. Treatments for antagonist ovarian stimulation were administered to 75 women under the age of 40, and 2-D power doppler ultrasound was used to guide the oocyte harvest. Patients were allocated into three groups according to the time of intrauterine infusion of PRP (no PRP, PRP at triggering day and PRP at ova pickup day). Power Doppler is utilized to measure endometrial thickness (ET), pulsatility index (PI), and resistance index (RI) of sub endometrial arteries, as well as serum levels of leukemia inhibitory factor (LIF). The mean endometrial thickness as well as serum level of LIF in PRP trigger group were higher, resistance index and pulsatility index were lower in a significant manner than control and PRP ova pickup group at all the days of study (triggering, ova pickup and embryo transfer days). These findings revealed that intrauterine PRP infusion on ova pick up day is associated with a significant increase in serum level of LIF that in turn enhances endometrial thickness for patients scheduled for intracytoplasmic sperm injection (ICSI).
Luteinizing hormone (LH) plays vital role for optimal follicular growth, maturation and is responsible for inducing ovulation, various protocols in ICSI cycles used to prevent the premature surge of luteinizing hormone. Currently, flexible GnRH antagonist protocols appears to be mostly used protocols. individualization of controlled ovarian hyper stimulation (COS) needs better timing of antagonist administration, assessing variation in LH levels during COS helps in determining appropriate initiation of inhibition .This prospective comparative observational analysis was made on one hundred twenty females with infertility who were undergoing ICSI flexible antagonist protocol at the “High Institute for Infertility Diagnosis and Assisted Reproductive Technologies/Al-Nahrain University” from October 2020 to April 2022, primary infertility was the with higher percentage (78.3%) than the secondary type. However; the main cause of infertility among the participant was male type (36.7%) fallowed by the combined one (20.8%). A highly significant difference was present in LH levels at all time points, with level decreasing from 4.99 ± 1.90 at baseline to 4.82± 2.15 at antagonist starting day to 1.63± 0.87 at trigger day and p value was > 0.001, no statistical difference in LH levels regarding the three time points between patients who get pregnant and non-pregnant women with pregnant patients having higher baseline lower antagonist starting day, and higher trigger LH levels 5.16 ± 1.87, 4.30 ± 1.72, 4.30 ± 1.72 respectively. Difference in LH level between cycle day 2 and the day of antagonist starting according to starting day of antagonist reveals that the basal LH values declined significantly (p= <0.001) until cycle day 7 of stimulation after which the basal LH start to increase and also with a high significant difference (negative values mean that the level decrease in comparison to the base line level).The comparable LH level in the current study reveals that earlier initiation of antagonist may serve no purpose and it is advisable to include the measurement of LH level to the criteria of antagonist initiation for better individualization and better outcome and to avoid un necessary earlier start of antagonist.
Polycystic ovarian syndrome (PCOS) is a group of clinically distinct endocrine conditions. Insulin resistance-related proteins are involved in PCOS pathogenesis. Irisin is a new myokine that functions similarly to adipokines. Irisin has been linked to insulin resistance and metabolic syndrome. There have been conflicting findings regarding irisin levels and their connection to hormonal and metabolic alterations in PCOS .This study aimed to compare the levels of irisin in follicular fluid and serum between women with PCOS and healthy women and correlate them with oocyte and embryo quality in women undergoing the ICSI cycle .Ninety infertile Iraqi women were enrolled, 46 women with PCOS and 44 weight-matched non-PCOS controls, in whom ICSI was done for them and evaluated the level of follicular fluid, serum Irisin, and insulin level with measure of Homeostasis Model of Assessment-Insulin Resistance (HOMA-IR) and correlate them to with oocyte and embryo quality. HOMA-IR, FF, and serum irisin levels in PCOS considerably increased. (P ≤ 0.01). In addition, there was a significant negative correlation between Irisin levels in follicular fluid and serum with MII oocyte, oocyte maturity rate, and grade 1 embryo. The serum, follicular fluid Irisin, and HOMA-IR were significantly higher in PCOS than in control cases, which may cause a reduction in oocyte maturation, embryo quality, and pregnancy outcome.
Women who have transvaginal oocyte retrieval feel nervous and have mild to moderate pain because a needle is used to poke holes in the vaginal skin and ovarian capsule to get the oocytes. The aim of this study is compare between general and spinal anesthesia effect on ICSI outcome . A cross-sectional study was conducted in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies/ Al-Nahrain University, from first April 2021 to the first of February 2022. Patients were randomized blindly at time of oocyte retrieval for types of anesthesia given: (50 women subjected to spinal anesthesia and 50 women subjected to general anesthesia). All females were subjected to flexible antagonist protocol. Also, all of them had the same surgical procedure by the same physicians. ICSI outcome was compared between the groups. The study showed there was non-significant difference (p>0.05) between the two groups regarding pregnancy rate with higher oocyte number in women subjected to spinal anesthesia than those who were under the general anesthesia although the difference was not statistically significant. The fertilized oocyte number was non significantly higher in spinal anesthesia group than the general anesthesia. So; spinal anesthesia and general anesthesia have comparable ICSI outcome.
Pregnancy-Associated Plasma Protein (PAPP-A) is a zinc metalloproteinase in the insulin growth factor system (IGFs) produced by the syncytiotrophoplast region of the placenta. It plays a critical function in the cleavage of IGFBP4. In the ovary IGFs, it regulates follicular and oocyte maturation, and steroidogenesis. While in polycystic ovarian syndrome (PCOS) Hyperinsulinemia and hyperandrogenemia it causes follicular environment changes and early ovulation resulting in lower oocyte and embryo quality in patients and this will decrease the success of pregnancy in women enrolled in the ICSI cycle. The present study aimed to assess the relationship of PAPP-A levels in serum and follicular fluid in women with PCOS and non-PCOS with oocyte and embryo quality in women undergoing ICSI cycle. 45 infertile Iraqi women were enrolled. Women with PCOS had to meet at least two of the three criteria set by the Rotterdam ESHRE/ASRMS criteria, the age of the included women ranged between 20-45 years. In non-PCOS patients, PAPP-A has higher level in serum and follicular fluid but without a statistically significant difference matching with PCOS group. In addition, there was no significant correlation between PAPP-A levels in serum and follicular fluid with oocytes and embryo characteristics. However, PAPP-A levels are higher in serum and follicular fluid in women with positive pregnancy but without significant differences. PAPP-A had no correlation with oocyte and embryo quality.
Infertile couples undergoing Assisted Reproductive techniques are really receptive to any intervention intended to enhancement their fertility. Successful implantation of an embryo requires a receptive endometrium, a good quality embryo, and embryo endometrial synchronization in all species. Pharmacological agents such as aspirin and sildenafil have been included in previous clinical trials to improve endometrial vascularity and perifollicular blood flow but the results were inconsistent. To evaluate the effect of oral low dose aspirin or vaginal sildenafil citrate on endometrial thickness, retrieved oocyte, degree of maturation, embryo quality and pregnancy test. This comparative prospective study was conducted on the 60 infertile women who participated in intracytoplasmic sperm injection (ICSI) cycles at the infertility clinic associated with the "High Institute of Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University / Baghdad/ Iraq." December 2020 to June 2022. In the present study the count of total oocytes retrieved was significantly higher in aspirin treated group in comparison to sildenafil treated group 12.5 ±5.3 versus 9.37 ±5.5 respectively. No significant variation in mean endometrial thickness among study groups. Regarding biochemical pregnancy test, positive results were obtained in 12 (40.0 %), 10 (33.3 %) in aspirin treated group, sildenafil treated group respectively and the difference was not significant. Aspirin treatment resulted in higher number of total retrieved oocytes, aspirin and sildenafil did not improve endometrial thickness, but they resulted in improvement of sub endometrial vascularity; however, biochemical pregnancy test was not affected by either pharmacological agent.
Inhibition of luteinizing hormone (LH) by gonadotropin releasing hormone (GnRH) antagonist may lead to suboptimal response during ovarian stimulation. In addition, several studies suggest that low level of LH is associated with lowered fertilization and implantation rate and increased early pregnancy loss rate. The aim of this study is to study the effect of adding low dose human chorionic gonadotropin (hCG - 200 IU), as an LH supplement, to recombinant follicle stimulating hormone (rFSH) in a GnRH antagonist cycles in women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment. Sixty-three infertile women undergoing IVF were randomly divided into two groups. One group was stimulated with the conventional stimulation protocol (rFSH alone), while the second group received 200 IU hCG in addition to rFSH in the late follicular phase (hCG + rFSH). Both groups' results including pregnancy rate, total dose of rFSH required, duration of stimulation, endometrial thickness, oocytes and embryos characteristic, serum hormone levels (Testosterone, Estradiol, Progesterone, and LH) and level of epidermal growth factor (EGF) were compared. The results show that pregnancy rate among the group of women who received the low dose hCG was higher than those who did not receive hCG. However, this difference did not reach statistical significance. Furthermore, other cycle outcomes and hormonal values were comparable between the two stimulation protocols.
In recent years increasing evidence proposed that local immune cells at implantation site have largely contributed to embryo implantation. The intrauterine infusion of activated peripheral blood mononuclear cells culture 2 days before embryo transfer can enhance the implantation. One of the methods used to evaluate the endometrial receptivity is by assessing the sub endometrial blood flow. A total of 67 infertile women (30) women receives intrauterine non-invasive insemination of peripheral blood mononuclear cells (PBMC) culture 2 days before embryo transfer representing the PBMC test group, and (37) women without receiving any cell as Non-PBMC group. The cultured PBMC are administered into the uterine cavity of the patients. 2 days later, embryos are transferred into the uterine cavity. Endometrial thickness and sub-endometrial blood flow measurements are taken for all cases on trigger and embryo transfer days. On embryo transfer day there was no significant difference (p = 0.770) in mean endometrial thickness between the PBMC group and Non-PBMC group. There was a significant difference (p< 0.001) in the mean resistive index; the level being lower in the PBMC group. Moreover, there was a significant difference (p< 0.001) in the mean pulsatility index. Regarding all enrolled women, the pregnancy rate of 25.4 %, the rate was higher in the PBMC group in comparison with the Non-PBMC group, 43.3 % versus 10.8 %, respectively and the difference was significant (p = 0.002). The use of PBMC culture can improve sub-endometrial. Endometrial Receptivity, ICSI, Fresh Embryo Transfer, Peripheral
Background : Bacterial vaginosis (BV)is a polymicrobial clinical syndrome re sulting from replacement of the normal hydrogen peroxide producing Lactoba cillus sp. BV is the most common lower genital tract disorder among women of reproductive age (pregnant and non-pregnant) and the most prevalent cause of vaginal discharge and malodor. Objectives: To study the role of asymptomatic bacterial vaginosis on outcome in patients undergoing ICSI. Patients and Methods: This prospective study was undertaken in the High Insti tute of Infertility Diagnosis and Assisted Reproductive Techniques / Al-Nahrain University. During the period from September 2017 – April 2018. A total of 100 infertile women .All of them were underwent controlled ovarian hyper stimula tion for intra cytoplasmic sperm injection cycle. Antagonist oocyte maturation used as ovulation induction protocol in all the cases. Assessment of oocyte mat uration and embryo quality is done to the all the cases, bacterial vaginosis smear was collected on the day of oocyte retrieval. B.V. smear diagnosis by Amsel’s Criteria and Nugent’s Scoring. Results: TThere was no significant difference in mean age of pregnant and non-pregnant ladies, 29.40 ±6.47 years versus 30.78±6.84 years, respectively (P= 0.333).In addition the frequency of women under 35 years age was compara ble in the two groups with no statistical significant difference, 34 (79.1%) versus 33 (73.3%), respectively (P = 0.528). Wide variation and non-normal distribu tion was the role followed by the duration of infertility concerning sub-fertile women participating in this study. However, no significant difference was ob served in the duration of infertility between pregnant and non-pregnant ladies, 6.00 (5.00) years versus 6.00 (5.00) years (P = 0.289). Comparison of basal hor monal levels between pregnant and non-pregnant groups revealed no significant difference despite the presence of some minor differences in mean hormonal levels between the two groups (P> 0.05). Comparison of oocytes characteristics between pregnant and non-pregnant ladies was conducted and revealed no signif icant difference in mean total number retrieved oocyte (TNO), ruptured oocytes (RO), abnormal oocyte (AO), germinal vesicle oocyte (GV), metaphase I oocyte and metaphase II oocytes (P > 0.05), table Mean total number retrieved oocyte (TNO), ruptured oocytes (RO), abnormal oocyte (AO), germinal vesicle oocyte (GV), metaphase I oocyte and metaphase II oocytes was 10.81 ±4.95 versus
Fertility is a fundamental function of reproductive health that is under the control of different hormones, cytokines, and growth factors. Transforming growth factor superfamily is a group of cytokines necessary for blastocyst implantation as they regulate endometrial stromal cell proliferation, and promote embryonic adherence to the receptive endometrium. Hyaluronic acid molecule (HA) is a long, unbranched polysaccharide that plays a delicate function in reproduction. Hyaluronic acid regulates gametogenesis, folliculogenesisi, and implantation, thereby was oroposed to improve the rate of successful pregnancy.This study is a prospective clinical trial that includes 80 infertile Iraqi women aged (18-40 years) scheduled for fresh ICSI program. The study sample had been divided into 2 groups based on the type of intervention performed on the day of oocytes retrieval. The Hyaluronic acid group and the control group. For each participant, twice measurement of the serum level of transforming growth factor were done , the first measure was done on day of oocytes retrieval, while the second measure was performed on day of embryo transfer. A comparative statistic was made to examine the effect of hyaluronic acid intrauterine wash on the serum level of transforming growth factor and pregnancy rate.Hyaluronic acid intrauterine wash exerts a significant reduction in transforming growth factor serum level, and a significant improvement in the pregnancy outcome compared to the control group.
Infertility is described as a couple's failure to conceive for at least a year, using unprotected sex. Even though all anesthetic chemicals have been revealed in the follicular fluids, general anesthesia is still used in many IVF centers for patients who want to get pregnant. This study included 60 infertile women randomly divided into two groups under general anesthesia. 30 patients were given a Ketamine dose of 0.5 mg/Kg and the other 30 patients were received Remifentanil dose of 0.5 µg/Kg for induction of anesthesia. We collected serum for Reactive Oxygen Species (ROS) levels assessment for all patients before and after starting general anesthesia. Routine ICSI procedures was performed on all participants, including clinical evaluation (history, examination, and investigation), controlled ovarian and ovulation stimulation, oocyte retrieval under general anesthesia, follicular fluid collection for postoperative anesthesia medication concentration (Remifentanil and Ketamine), oocyte stripping, oocyte maturity assessment, intra-oocyte sperm injection into mature cells (MII), fertilization and division evaluation and embryo categorization, embryo selection and transfer, luteal phase support, and beta-hCG determination. ROS concentrations were compared between the Remifentanil and Ketamine groups. There was no significant difference in embryo features between Remifentanil and Ketamine, indicating that neither one is superior to the other in this regard. When Remifentanil or Ketamine was taken, there was no significant difference in ROS levels in serum or follicular fluid.
The effect of maternal body mass index (BMI) on fertility outcomes in women undergoing in vitro fertilization/intracytoplasmic sperm injection cycles has been extensively evaluated and the results of these studies have shown a lot of controversial issues. Folate is a naturally occurring type of vitamin B9 crucial for reproductive health. 65 infertile couples were subjected to intracytoplasmic sperm injection cycles. Both primary and secondary types of infertility were involved, with different causes. The mean plasma folate of all infertile women was 12.71±6.52, with pregnant 11.60±5.57 and non-pregnant 11.74±8.80; with no significant difference in mean plasma folate between them. Moreover, the means of follicular fluid folate of all infertile women, pregnant women, and non-pregnant women were 8.00±5.39, 7.84±4.68, and 8.39±6.19 respectively. There was no significant statistical difference in mean follicular fluid folate between pregnant and non-pregnant women (p=0.719). Also, both plasma folate and follicular fluid folate were not significantly correlated to oocyte and embryo characteristics. Although plasma folate was higher in obese than normal and overweight women, the difference did not reach statistical significance. It appears that the correlation among maternal BMI, folate level and fertility outcomes in women undergoing intracytoplasmic sperm injection cycles are still controversial and much research work is needed to figure out such complex interaction among these variables.
Background: Chemotherapy and radiotherapy can destroy or severely reduce spermatogenesis and thereby jeopardize fertility in the long term. There is still no medical treatment that guarantees fertility preservation after chemo- and radiotherapy. Now with recent improvement of assisted reproductive technologies (ART) and possibility of using testicular spermatozoa or epididymal spermatozoa at in vitro fertilization (IVF) by intracytoplasmic sperm injection (ICSI), cryopreservation of testicular tissue is an option in fertility preservation for males who will lose spermatogenic cells as a result of chemoand radiotherapy and for males with azoospermia. Objectives: This study is an attempt to evaluate oxidative DNA damage in the mice testicular tissue after cryopreservation/thawing cycle when using different types of cryoprotectants as well as to investigate the changes that occur in mice testicular tissue after cryopreservation/thawing cycle as a model for human being. Methods: Fifty mature fertile male mice between 8- 12 weeks old were used in the current study. For each mouse, one testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) without cryopreservation (control group). The other testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) after six weeks of cryopreservation using different types of cryoprotectants (cryopreserved group). Results: The microscopically observation of slides obtained from cryopreserved testis differs according to the type of cryoprotectant (glycerol, 1,2 propanediol and dimethylsulfoxide), in the dimethylsulfoxide group, tissue sections showed no major differences versus control group, but in the histological sections obtained from tissue cryopreserved by glycerol as cryoprotectant showed moderate morphological and structural changes as compared with the control group. While, the tissue samples subjected to 1,2 propanediol as cryoprotectant displayed the severest morphological and structural changes as compared with the control group. Immunohistochemical results of the present study showed a highly significant (P<0.001) increase in oxidative DNA damage in the cryopreserved testis (46.78%, 63.45% and 32.59% represent cryoprotectant glycerol, 1, 2 propanediol and dimethylsulfoxide, respectively) compared with control group (19.27%) after six weeks of cryopreservation. Conclusion: From the results of the current study, it was concluded that there was alteration in testis histology after cryopreservation/thawing cycle and increase the level of oxidative DNA damage after cryopreservation/thawing cycle. As well as dimethylsulfoxide cryoprotectant provide good protection for testis histology and DNA.
Kisspepetin is considered as a key regulator of reproduction, through its direct role in controlling the hypothalamic pituitary gonadal axis necessary for controlling the ovulatory process and indirectly by impacting glucose heamostasis and metabolism. We aimed to examin the correlation between body mass index and level of Kisspeptin in serum and follicular fluid. This study is a cross sectional study done on (80) infertile Iraqi patients aged (18-40 years) scheduled for fresh intracytoplasmic sperm injection cycle. This study is conducted in the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies of Al-Naharin University, from September 2020 to May 2021. For each participant, the GnRH antagonist protocol was used to hyperstimulate the ovaries, and samples from serum and follicular fluid were obtained at day of oocytes retrieval for measuring Kisspeptin level using enzyme linked immunosorbent assay (ELIZA) technique. Serum and follicular fluid Kisspeptin were insignificantly increasing with the increase in the BMI (p value =0.406) (p value = 0.352) respectively. Serum and follicular kisspetin were positively and significantly correlated (r = 0.536, p value= 0.000). Serum and follicular fluid kisspeptin were numerically increasing with the body mass index of infertile women, although this increament is statically insignificant, serum kisspeptin can be used as a mirror image to follicular fluid kisspeptin as there was a statistical association between them in this study.
Background: Since 1999 the World Health Organization (WHO) recommended the assessment of sperm morphology by Kruger strict criteria (KSC). The manual of WHO contains the cut off values of sperm morphology measured by KSC for normal subject and patients treated by in vitro fertilization (IVF) and intra- cytoplasmic sperm injection (ICSI) without any information regarding intrauterine insemination (IUI). Objectives: The objective of this study is to achieve two major goals that are:1- Assessment of sperm morphology by applying KSC, using two different biological stains, named commercially Testsimplets (TS)pre-stained slides and Sperm-O-Scan (SOS)stain 2- To have a cut off value for morphologically normal sperm that resulting into a successful IUI outcome. Materials and Methods: Fifty five unselected couples attending the Institute of Embryo Researches and Infertility Treatment were included in this prospective study from December 2007 till August 2008. The mean age of male partner was 31.7±1.87years with an average duration of 6.25±1.65years of infertility. The women were prepared for IUI. All the males were subjected to semen analysis with in vitro sperm activation using layering swim-up technique. Assessments of the morphologically normal sperm (MNS) of the samples were performed by using both, High power field and KSC by using TS and SOS stains before and after the activation. Correlation of the positive IUI results and MNS by KSC waw recorded denoting the cut off value for successful results. Results: The mean of morphologically normal sperm applying KSC with the use of SOS stain before in vitro sperm activation was 16.10% and after activation was 37.85% and by using TS stain before in vitro sperm activation was 15.14% and after activation was 37.09%. No significant (P>0.05) differences was detected between the results of MNS of the two stains. The result of IUI was 11 positive pregnancies out of the 55 cases. The mean of MNS using KSC for these positive cases was 11.58% and 12.84% for SOS and TS stains, respectively. Conclusions: The cut off value of MNS by applying KSC and using two different biological stains for a successful IUI program was 11.58% by SOS stain and 12.84% by TS pre-stained slides. Although the study found no statistical differences between the two stains, TS found to be easier, simpler and less time consuming technically than SOS stain.
It is well known that oocyte quality is essential in the determination of the developmental potential of the fertilized oocytes. The oocyte developmental potential decreases in all species with increasing age. Use of serum and follicular fluid concentration of GDF9 as biomarkers of oocyte and embryo quality and their relation to pregnancy rate. Eighty-eight women were included in this study are selected from those undergoing intra-cytoplasmic sperm injection. Positive pregnancy was achieved by 14 women. The difference in mean serum and follicular GDF9 between pregnant and non-pregnant women was not significant. MI oocyte count was not significantly correlated to serum and follicular GDF9 (p > 0.05). MII oocyte count showed a non-significant correlation to serum and follicular GDF9 (p > 0.05). Grade 1 embryo count showed a non-significant correlation to serum and follicular GDF9. Grade 2 embryo count showed a non-significant correlation to serum and follicular GDF9. Also, grade 3 embryo count showed a non-significant correlation to serum and follicular GDF9. The current study revealed that serum and follicular GDF9 could not be used as indicators for oocyte maturity, embryo quality, or pregnancy rate.
Gonadotropin-releasing hormone antagonists are one of the GnRH analogs that were used in assisted reproductive technologies to produce prompt downregulation of pituitary gonadotropin secretion. During conventional antagonist protocol (CAP), exposure to high LH and E 2 occur that have the potential to worse clinical reproductive results. So the downregulation of pituitary secretion for short period during early follicular phase will result in synchrony in follicular developments and this will improve mature oocytes and total embryos numbers. 44 women as normal responders undergoing ICSI-ET cycles were randomized into two groups. The conventional group (CG) (n 30), gonadotropin started from menstrual cycle day 2 or 3 and continue until hCG trigger day, flexible protocol in which GnRH antagonists administered with follicular size (13-14 mm). In the sandwich group (SG) (n 14), a GnRH antagonist was administered for three days in which GnRH antagonists administered with follicular size (13-14 mm). Gonadotropin started from menstrual cycle day 3 and continue until hCG trigger day. Retrieved and MII oocyte mean numbers were significantly higher in SG than in CG ( P = 0.006, and 0.025), respectively. Embryos and frozen embryos mean total numbers were significantly higher in SG than in CG ( P = 0.004). SG patients have a higher pregnancy rate of 9/14 (64.3 %) than CG 12/30 (40.0 %) although not significant ( P =0.057). Early and short GnRH antagonists proved improvements in synchronization of follicular development, retrieved mature oocytes numbers, total embryos, frozen embryos, and pregnancy rates.
Placenta growth factor is, VEGF family member, a Pleiotropic Factor affects a variety of cell types and regulates various biological responses. PlGF is similar in structure and function to VEGF, and it amplifies VEGF's angiogenic actions. Ovulation stimulation during cycles of in vitro fertilization (IVF) for the management of infertility results in ovarian hyperstimulation syndrome (OHSS), an iatrogenic side effect. Considering potential roles of PlGF and its receptor (sflt-1) in angiogenesis, the association of these factors with OHSS among study women during controlled ovarian stimulation have evaluated in the present study. Comparative cross-sectional study including of 60 women who go through controlled ovarian stimulation. On oocyte retrieval day, their serum and follicular fluid were taken. The concentrations of PlGF and sFlt-1 were assessed using ELISA .Eighteen patients presented with ovarian hyperstimulation syndrome (OHSS) and 42 patients were no OHSS. There was significantly higher serum PIGF in women with OHSS patients as compared to women with no OHSS, 141.4 ± 11.8 versus 91.9 ± 5.4 respectively (p<0.001). Furthermore, Follicular fluid PlGF there was significantly higher in women with OHSS patients as compared to women with no OHSS, 163.4 ± 7.2 versus 91.1 ± 5.5 respectively (p<0.001). Additionally Follicular fluids PlGF/sFlt ratio significantly higher in women with OHSS patients as compared to women with no OHSS, 0.034 ± 0.01 versus 0.026 ± 0.01 respectively (p<0 . 001). On the contrary there was significantly lower serum and follicular fluid sFlt in ovarian hyper-stimulated patients These statistics show that serum and FF PlGF and PlGF to sFlt ratio higher in women with ovarian hyperstimulation syndrome (OHSS) patients as compared to women with no OHSS. So, the PlGF may be played an important a role in angiogenesis dysregulation.
Poor ovarian responders (PORs) are characterized by decreased the response of the ovaries to stimulation. Nowadays, using soft protocol of low doses of Gn with/without oral compounds such as letrozole was accepted as a good option. The endometrium receptive parameters include sub endometrial resistance, pulsatility index and endometrial thickness. Eighty infertile POR women divided to two equal groups and undergoing IVF/ICSI antagonist protocols. One group underwent soft protocol using letrozole 2.5mg twice daily for 5 days starting from cycle day 2-3 overlapped low-dose Gn (225 IU) from cycle day 4-5. Other group received high dose Gn only (450 IU) from cycle day 2-3. Both groups continued stimulation cycle until the day of trigger using the flexible antagonist protocol. Estimation of values of sub endometrial RI and PI with endometrial thickness by 2 Color Doppler ultrasound was performed at day of trigger, OCP and ET. There was no significant difference for endometrial thickness at day of trigger, OCP and ET. There was significant difference in means of sub endometrial RI and PI at day of trigger, OCP and ET (P>0.05). This study showed evidence in favor of soft protocol for POR regarding pregnancy rates in spite of the statically difference in means of sub endometrial RI and PI associated with much less cost effectiveness.
Various types of anesthetic techniques were described for oocyte recovery. Different anesthetic agents have been used with different effects on oocyte quality and embryonic development and studies on the potential toxicity of general anesthesia ended up with conflicting results. Many experiments as well as human studies documented the existence of anesthetic agents in the follicular fluid (FF) with possible adverse effects on the oocyte quality and subsequent embryo development. Ketamine and remifentanil were used for the studied groups to compare oocytes and embryos characteristic of infertile women during oocytes retrieval under general anesthesia: one group receiving ketamine, midazolam, and propofol and the other group receiving remifentanil, midazolam and propofol. 60 infertile females were undergoing intracytoplasmic sperm injection categorized in two groups, group I received midazolam, remifentanil and propofol; whereas, group II received midazolam, ketamine and propofol. There was highly significant difference in mean abnormal oocyte between ketamine and remifentanil groups. Grade III embryos were significantly limited to ketamine group. FF ketamine and remifentanil were not significantly correlated to any of oocyte characteristics or embryo characteristics. Anesthetic agents, ketamine and remifentanil, were detected in the FF of infertile women and within recommended doses. Ketamine resulted in adverse effects on oocyte and embryo quality in comparison with remifentanil represented by greater percentage of abnormal oocytes and that bad quality embryos were limited to ketamine group.
Background: In vitro Fertilization (IVF) is an important alternative for infertile couples, there is an increase in the number of children conceived by IVF worldwide. Objective: The aim of this study was to assess the risk of birth defects and adverse outcome among infants and children conceived by in vitro fertilization in Sulaimani-Iraq. Patients and Methods: This case-control study was carried out on 160 children, 75 were product of IVF(using ICSI) and 85 were product of natural conception. Data collected regarding mother age, father age, duration of infertility before pregnancy with the index child, any complication during pregnancy, age of the child, gender, mode of delivery, gestational age, weight at delivery, history of neonatal admission and it’s cause, admission to hospital after neonatal period, and history of surgical operation. All children were examined for any gross anomaly, cardiovascular, respiratory, gastrointestinal, skeletal, urological and neurologic abnormality. Results: IVF was a risk factor for prematurity (OR, 7.844 95% CI 3.025 - 20.339) , LBW (OR, 4.645 95% CI 2.038 -10.587), Congenital anomaly (OR, 1.362 95% CI 0.720 - 2.576), predominance of male gender (OR, 1.340 95% CI 0.719-2.498), neuro-developmental delay(OR,1.333 95% CI 0.672-2.646), admission to NICU (OR,1.703 95% CI 1.052-2.758 ), C/S delivery (OR, 1.972 95% CI 1.507-2.581), and childhood hospitalization (OR, 1.193 95% CI 0.838-1.696), also there was statistically significant association with multiple pregnancy. Conclusions: IVF carries increased risk of LBW, prematurity, congenital anomaly, neuro-developmental delay, admission to NICU, childhood hospitalization, C/S as a mode of delivery and predominance of male gender.
In human-assisted reproduction both partners need to be evaluated during counseling because causes of infertility are due to male factors and female factors. some body hormones play important role in fertile women and cortisol one of these hormones which is essential for normal ovulation, cortisol usually increases during the follicular phase of the menstrual cycle, play a role in the early stages of follicular development by their interaction with aromatase activity and granulose cell functions. To investigate the correlation of cortisol hormonal level in serum and in follicular fluid that relationship with age in women under Intracytoplasmic sperm injection outcome. Its randomized prospective clinical trial study performance conducting in the Higher Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University, Baghdad-Iraq, during the period from November2021to June 2022. a total of Sixty-three Infertile Iraqi women, who planned to inter intrasytoplasmic sperm injection program. The blood sample and follicular fluid sample obtained from all infertile women inter intracytoplasmic sperm injection program to measure cortisol hormonal level at specific days of this study. There was increase in cortisol level in women aged between (30-40) years, So there was significant finding between cortisol level and this age group and there was increase in cortisol level in women aged < 30 years at the same day and no significant difference in cortisol level in women age more than 40. Significant differences in cortisol level appear in group of women age between (30-40) years, also the same result appears in group of women age more than > 40 years and no significant differences in cortisol level in women aged < 30. No significant differences in cortisol level in all age women group (30-40) years, <30 years ,and the women age > 40 years.
Background: The sperm plasma membrane activity and absence of antisperm antibodies in infertile patients ejaculate significantly associated with fertilization potential of human spermatozoa. The sperm viability detected by hypoosmotic swelling test (HOST) and autoimmune infertility detected by antisperm antibody assay presented diagnostic and prognostic tools for male infertility factors. Objective: This study was devised to compare the results of hypoosmotic swelling test and antisperm antibody assay for normozoospermic men and infertile patients affected by autoimmune infertility. Methods: Thirty semen samples were collected by masturbation and prepared by direct layering technique. Hypo-osmotic swelling test was performed by mixing 0.1 ml of semen with 0.9 ml of a 150 mOsm/L NaCl as a hypo-osmotic solution and direct immunobead assay were used to determine the presence of Antisperm Antibodies (ASA) bound on sperm surface. However, sperm concentration, sperm motility, and normal sperm morphology were evaluated according to World Health Organization (WHO) criteria and subjected to HOS test and ASA assay. The sperm prepared and incubated for 30 minute in 5% CO2 at 37°C after in vitro sperm preparation. Results: The results of the present study reported that the percentage of HOS test score in antisperm antibodies positive sperm samples (52.30 ±5.2) was significantly lower than that noticed in antisperm antibodies negative (70.30 ± 3.4) sperm samples. Conclusion: It was concluded that the use of HOS test as a simple and diagnostic and prognostic tools for identify male infertility. Further studies are suggested to assess the effect of ASA on sperm plasma functional integrity after ICSI and in vitro fertilization and embryo transfers (IVF-ET).
For successful implantation, a high-quality embryo, receptive endometrium, and tight communication between the two are required. The lack of uterine contractions during embryo transfer is one of the characteristics that greatly affects endometrial receptivity. This is because it has been observed that ET may cause uterine contractions, which may be harmful to embryonic apposition and endometrial blood flow .The aim of this study was to assess how magnesium sulfate administration on the day of embryo transfer improve sub endometrial blood flow, endometrial receptivity, and increase implantation rate and clinical pregnancy rate . Eighty infertile women undergoing ART cycles participated in this prospective. Recombinant FSH (Gonal-F) was used in a controlled ovarian hyperstimulation technique. The first group A (n=40) got a 4gm infusion of magnesium sulfate, while the second placebo control group B (n=40) received 200ml of 5% glucose water as a placebo, respectively. While there were no significant changes in body mass index (BMI), endometrial thickness, number of transferred embryos, or embryo transfer day, significant variations were seen in patient mean age (p<0.001) and mean number of grades I embryo (P=0.003). It was significantly different after magnesium sulfate administration with the lowest value in magnesium sulfate group (P=0.024; P <0.001 and P <0.001 respectively). Implantation rate and clinical pregnancy rate both increased after drugs administration although it was non-significant (P= 0.106 and P= 0.348 respectively). It appears that magnesium sulfate injection before to embryo transfer can significantly increase sub endometrial vascularity and endometrial receptivity and improving implantation rate and clinical pregnancy rate non significantly in ICSI cycles by lowering uterine muscular
Background : Although progress was made in an in vitro fertilization (IVF) tech niques, the majority of transferred embryos fail to implant. Morphology embryo scoring is the standard procedure for most of IVF centers for choosing the best embryo, but remains limited since even the embryos classified as (top quality) may not implant. Determining molecular mechanisms of human embryo implan tation is an extremely challenging task due to the limitation of materials and significant differences underlying this process among mammalian species. L-Se lectin and its ligand carbohydrate have been proposed as a system that mediates initial adhesion of human blastocysts to the uterine epithelia. Objectives: Quantitative evaluation of soluble L-Selectin molecule in an in-vitro culture media used for in vitro fertilization. Determine cut-off value of soluble L-Selectin quantity to be as qualitative predictor of successful implantation Patients and Methods: This prospective study was undertaken in the High In stitute of Infertility diagnosis and assisted Reproductive Technologies/ Al-Nah rain University/ Baghdad/ Iraq, during the period from September2017 to April 2018.A total of 74 infertile women were underwent controlled ovarian hyper stimulation for intracytoplasmic sperm injection cycle. Flexible antagonist pro tocol was used as ovulation induction protocol in all the cases .culture media L-Selectin level were measured on the day of embryo transfer by using Enzyme linked immune sorbent assay (ELISA) for all cases . Comparison in culture me dia L-Selectin was done to all cases. Results: There was no significant difference between pregnant and non-pregnant groups in age , BMI, and level of basal hormone FSH, LH , AMH and E2 in the day of hCG injection, and significant difference in the level of L-Selectin in pregnant compared to non-pregnant groups. Conclusion: Level of L-Selectin concentration in embryo culture media can be used as predictor of ICSI success
Azoospermic patients have the option of undergoing in vitro fertilization procedures, specifically intracytoplasmic sperm injection (ICSI), for which sperm is retrieved from the patient's own testicles and then injected directly into the oocyte. The aim of the study: evaluate the role of repeated sperm extraction by enzymatic and mechanical mincing techniques in azoospemic patients. A total of 32 males with non-obstructive azoospermia participated in the research. Testicular sperm extraction (TESE) was performed under spinal or general anesthesia after a comprehensive history, physical examination, and investigations. After that, the biopsies underwent through a series of mincing processes, both mechanical and enzymatic. Patients gave information including age, body mass index and previous biopsy attempts. In both enzymatic and mechanical methods there was no significant higher sperm recovery in patients with history of previous testicular biopsies (66.7% vs. 38.5%; p=0.209) and (66.7% vs. 30.8%; p=0.102) respectively. Patients who had undergone prior testicular biopsies had marginally higher sperm recovery rates using both enzymatic and mechanical techniques