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.
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
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Background: Oocyte vitrification is a promising technique. The choice of appropriate types and concentrations of cryoprotectants is essential for the success of the oocytes vitrification. Objective: This study aimed to investigate the effect of vitrification on viability and morphology of oocytes and to compare the effect of several cryoprotectants on the viability and morphology of oocytes during vitrification and post-thawing. Methods: The sheep ovaries were collected from the local abattoir. Only normal and viable oocytes were included in this study. By using cryotop, immature oocytes that were viable with normal morphology were vitrified with 15% DMSO and 15% EG supplemented with 0.0 M, 0.25M, or 0.5 M of either sucrose or trehalose as control and treated groups. Oocytes viability and morphology were assessed post-aspiration and post-thawing. Results: Fromthe results of the present study, the percentage of post-thawing normal and viable oocyte reported with the use of 0.5M trehalose and EG in vitrification solution (VS) significantly)P<0.05) higher than the percentage of post-thawing normal and viable oocyte reported with use of 0.25 M trehalose and EG. Conclusion: Vitrification is simple technique and easy to perform but it needs some experience to prevent any oocyte loss during vitrification and thawing processing. The use of 0.5 M of either sucrose or trehalose in vitrification solution improves the percentage of post-thawing viable and normal oocytes.
Background: Speramax® was found to play an important role in sperm function characters and males reproductive performance with no studies on its effects on the oocyte maturation and embryonic development in females. Objective: The goal of the present work was to examine the effect of Speramax ® on oocyte maturation ,ova quality , embryonic development and live birth using mice as a model for mammals . Materials and Methods: In this study, Speramax ® was administrated orally for 1, 2 and 4 weeks . One hundred and ten female mice were randomly divided into four groups , the first group was superovulated(SUO) while the second group was treated by Speramax® with SUO and the third group was spontaneously ovulated (SPO) and treated with Speramax ® and the forth group was spontaneously ovulated (SPO ) and served as control group .Another forty-eight female mice were used for the determination of the number of live births . Results: The results indicated that treatment with Speramax® showed a positive effect on oocytes maturation in vivo. There was a highly significance ( p˂ 0.0001) improvement in the number of mature oocytes following treatment with Speramax® in SPO and SUO mice compared with SPO and SUO mice not treated with Speramax® . The percentage of embryonic development after 24 and 48 hours of mating in treated groups with Speramax® was significantly ( p˂ 0.05) higher than SPO and SUO mice .The study showed that the quantity and quality of embryos obtained from the treated groups were superior to that of the untreated group . Conclusions: It was concluded that the Speramax® greatly improved oocyte maturation, early embryonic development and embryo grading quality embryos with an increase in the numbers of mice live births.
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.
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.
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
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.
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.
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.
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.
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.
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.
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.
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.
Background: Many studies found that the Glycyrrhiza glabra (G.glabra) showed a positive effect on the activation of sperm in vitro specifically improve the activity of progressive movement. However, it was not known the impact of this stimulant on the certain sperms parameters in vasal obstructive mammals. Objectives: This experiment was designed to examine the possibility of fertilization ova by epididymal sperm of vasectomized mice activated by G. glabra and study the early embryonic development, as a model for obstructive azoospermia. Methods: Glycyrrhiza glabra aqueous extract (0.3 mg/ml culture medium) was used for in vitro direct sperm activation technique. The superovulated female mice (SUO) were divided into two groups, the first group; the insemination of flushed ova was performed with epididymal sperm of vasectomized male mice activated by adding30% of G. glabra to the culture medium (treated group).While the second group,the ova were inseminated by epididymal sperm of vasectomized male mice activated by G. glabra - free medium (control). Results: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams -F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively). Conclusion: This study showed a highly significant (P <0.01) improvement in certain sperm function parameters i.e the sperm forward movement, morphologically normal sperms (MAS). There was a significant (P<0.05) increase in fertilization rate (FR) of SUO mice oocyte using 30% G. glabra medium(75.84%) compared to G. glabraFree medium (Hams-F12 medium) alone (54.43%).By adding 30% G. glabra to the medium, the number of 3-cell and 4-cell embryonic stage of SUO mice was significantly (P <0.05) higher than that of grouping G. glabra- Free medium mice embryo(62.10% and 65.78% vs. 58.88 and 66.29%, respectively).
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.
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.
Human peptidoglycan recognition protein-1 (PGLYRP‐1) protein molecule activation in the semen of infertile patients has been suggested to induce a negative impact on seminal fluid analysis. This study aimed to evaluate Human Peptidoglycan Recognition Protein 1 for indication of bad seminal fluid analysis parameters. One hundred seventy-six infertile patients were included in this study. Seminal fluid analysis and Endtz test were done; Human Peptidoglycan Recognition Protein 1 in seminal plasma using enzyme-linked immune-sorbent assay technology PGLYRP‐1 is evident in seminal plasma of infertile men and statistically different according to the type of infertility. The concentration of PGLYRP-1 in the semen is directly proportional to the presence of leukocytes (Endtz test), which is reflected negatively on the quality of the semen. To the best of our knowledge, this study has not been done in Iraq or elsewhere. The presence of PGLYRP-1 protein in the semen may be evidence of leukocytospermia that may be related to bad quality for semen across infertile men with higher sensitivity than the Endtz test.
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.
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).
Background: The immune infertility caused by anti-sperm antibodies (ASAs) represented about 1020%- of infertility among the couples, the ASAs interfere with sperm parameters such as the sperm motility and sperm ability to penetrate cervical mucus, sperm-oocyte binding, and fertilization and embryo developments. Objectives: The present study designed to assess semen analysis, presence of ASAs and DNA fragmentation index as well as correlation within these parameters in normzoospermic Iraqi subjects Patients, Materials and Methods: A total number of Iraqi subjects (116) with range of age (2051-) years and their mean duration of infertility (4.70 ± 2.77). Seminal fluid for Macroscopic and Microscopic assessments done according to WHO 2010 criteria. The mixed agglutination reaction (MAR) test used to assess theirASAs in semen (direct method), seminal plasma and serum (indirect method); for the both IgG and IgA classes of antibodies and their distribution among different parts of spermatozoa as percentage. Results: The mean of IgG using direct method was (23.881.75±) and for indirect detection method the mean was (27.412.41±) with no significant difference (P<0.05) between the two methods. While the direct method for IgA detection mean (14.46 ±1.76) and the mean for indirect method of detection (6.860.39±) and there was significant difference (P>0.05) between the two methods. In addition to that, the distribution of IgG and IgA detected by direct method on sperm parts showed no significance difference except for the sperm tail; while the indirect method for IgA and IgG showed significant difference in distribution on sperm mid-piece. Conclusion: The correlation among sperm parameters and Immunoglobulin using direct and indirect methods showing variations in significance also in the correlation between the direct and indirect methods according to distributions on sperm surface
Mitochondria play a significant key in sperm metabolism as well as power generation. The mitochondria nucleolus is essential for several sperm activities. such as sperm capping, hyperactivation, and acrosome reactions, and oocyte fusion, mitochondrial membrane potential can be used to study sperm mitochondrial activity, the mitochondrial integrity of sperm is evaluated by examination of mitochondrial membrane potential MMP, which may be achieved by using fluorescent dyes like JC-1 probe is composed of 5,5,6,6-tetrachloro-1,1,3,3-tcarbocyanine iodide. To study the correlation between mitochondrial membrane potential Red MMP and sperm motility . This study included 42 male samples who were separated into two groups. (21 normozoospermic, 21 asthenozoospermia) through the period of attendance at the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, and Um Al-Banin Center for Management and in vitro fertilization the collected semen was obtained and seminal fluid analysis was assisted according to (WHO,2021). fluorescent staining of the sample prepared is done by using an MMP Assay Kit (with JC-1) was performed. A positive significant correlation between sperm motility percentage and red MMP intensity (p < 0.001). Mitochondrial activity plays an essential function in sperm motility and male fertility Male infertility, Sperm Motility, Mitochondrial Membrane Potential, MMP Assay Kit (with JC-1).
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
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.
Background: Recently the reproductive hormones, Anti-Mullerian hormone (AMH), activin A, and follistatin (FS) have been found to play an important role in folliculogenesis, oocyte maturation and corpus luteum function by changing the pattern of granulosa cell expression which in turn affects the success of fertilization potential. Objectives: To examine the relation of in vivo concentrations of AMH, activin A, and FS hormones on the ovaries status in the phases of menstrual cycle and to elucidate the relationship of these hormones with pregnancy rate following intrauterine insemination(IUI). Materials and methods: Seventy seven infertile couples were recruited from infertility clinic population at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies through the period from September 2011 to May 2012. Those infertile couples were divided to 3 groups according to the infertility cause (male factors causes, anovulatory causes and unexplained causes of infertility). Measurements of AMH, activin A, and FS hormones levels were done at cycle day (CD25-). Then measurements of activin A, and FS hormones levels were done only at preovulatory cycle(CD1214-) when the size of the Graffian follicle range between 1824-mm and at least one follicle was present and before hCG injection and at day 28 of the cycle (CD28). IUI was performed 3640- hours after hCG injection then measurement of the three hormones was performed after successful IUI. The result of successful pregnancy was recorded and statistical relation with AMH, activin A and FS hormones was assessed. Results: There was a positive significant correlation between endometrial thickness (ET) and pregnancy rate and the best ET to predict pregnancy is more than 7mm. There was no significant relation (P>0.05) between AMH level and pregnancy rate. A significant correlation was found between follistatin levels in preovulatory (CD 1214-) with a cutoff value 0.20ng/ml to predict pregnancy. There was a significant relation between activin-A level and pregnancy rate with a predictive cut off value 397.5 ng/ml on (CD 28) only. Conclusions: Activin-A can be regarded as a biomarker candidate for diagnosing very early pregnancy at luteal phase following IUI. Follistatin preovulatory (average CD13) has the most predictive value for pregnancy following IUI.
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.
Back ground: Optimal IVF and embryo culture conditions depend on the composition of culture media being the most important determinants of successful in vitro interaction of the gametes and subsequent embryo development. ATP is fundamental factor to maintain the life, by providing energy, and controlling the cell function and metabolism. Objective: To investigate the effect ATP supplied to culture medium on fertilization rate and in mice as model for human being. Materials and Methods: The female mice (90) were superovulated using superovulation program (SOP) to produce large number of oocyte were divided into three groups, G1 inseminated with vassal sperm in vitro alone (control) , G2 inseminated with vassal sperm in vitro using SMART enriched with ATP(1.25mM). While, G3 inseminated with vassal sperm in vitro using SMART medium enriched with ATP (2.5mM). Mouse spermatozoa were collected from both vas deferens. Then sperm parameters were assessed after 30 min. IVF technique was performed for 3 groups. Results: The results of the present study appear, significantly increment (P<0.05) in the IVF (%) was observed when using 2.5 mM ATP as compared to 1.25 mM ATP and control group. Conclusions: The result showed that addition of high ATP concentration (2.5 mM) enriched to the culture medium improved percentage of in vitro fertilization.
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
Background: Unlike sperm production in men which is continuous, women are born with all their eggs and they do not produce any more. Technique to bank eggs would allow girls and women to have the same reproductive options as men when faced with a serious disease and the long term complications of chemotherapies that threatens to destroy their eggs. Objective: The aim of this study is to improve the efficiency of existing ovarian cryopreservation, by following the most recent method named needle immersed vitrification (NIV) beside the slow freezing method. In this review we discussed also the historical description of the methods used, freezing and vitrification of mature and immature oocytes, this include comparison of principles, procedures and results as reported in the literature. Materials and Methods: A total of 285 fragments of human ovarian tissues were isolated and cryopreserved from 36 women aging 26 - 55 years old in the Reproductive Unit, Lubeck-Germany. Ovarian tissue strips of 1-3 mm thick of ovarian cortical biopsies obtained from patients. The collected human ovarian cortex fragments were assigned to two different cryopreservation procedures, slow-freezing and NIV. On the other hand, a total of 80 patients (100 cycles) with polycystic ovaries (PCO) were underwent in vitro maturation (IVM) treatment. The number of inspirated immature oocytes was 536. Vitrification of immature oocytes were carried out. Results: Successful freezing and storing were performed on 285 fragments of ovarian cortex tissue from 36 patients by two cryopreservation protocols as mentioned before but these tissues do not thaw and use yet. Concerning the IVM, the percentage of maturation, fertilization, survival, embryo transfer, and pregnancy rates were 55.2%, 44.5%, 96.2%, 88.1%, and 10.6%, respectively. Conclusion: Recent advances in freezing technology, modifications of conventional protocols used and continuing optimization of vitrification have efficiently improved the method. These results may lead to increase the possibility of preserving fertility by cryopreservation of ovarian tissue. Ovarian tissue banking can offer hope for cancer patients who want to safe guard their fertility against sterilizing chemotherapy and radiotherapy. Immature oocytes can be successfully isolated from the oophorectomy specimen regardless of the day of menstrual cycle, and undergo IVM and cryopreservation.
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.
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.
Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.
Background: Fibrillin- 3 is a recently discovered hormone.However, there are no studies till now regarding the role of fibrillin-3 hormone on fertilization capacity in infertile pregnant and non-pregnant women. Objectives: The goals of this study are concentrated on the fertilization capacity parameters by examining the effect of in vivo concentration of fibrillin-3 hormone on the ovarians status at different menstrual cycle phases and to elucidate the correlation between the levels of this hormone with pregnant and non -pregnant women following Intrauterine insemination (IUI) outcome or through natural pregnancy. Methods: Sixty infertile couples were involved in this prospective study. The couples were divided according to the type of insemination and ovulation induction program into two groups Group(1): Thirty spouses were subjected to induced ovulation stimulation by using clomiphene citrate(CC) and recommended for natural coitus. Group(2): Thirty couples, the females were induced ovulation stimulation by CC, Gonal F-751U /ml and Fostimon 50IU/ml with IUI. Measurements of Fibrillin-3 with other reproductive hormones were done at different phases of menstrual cycle and for pregnant and non-pregnant women following natural coitus and IUI. Results: The mean of fibrillin-3 hormone level at day 2 of menstrual cycle was 3.7ng/ml (ranged from 3.50 to3.91) in the two groups. The mean of fibrillin-3 hormone at cycle day 13 was 6.6ng/ml( ranged from 5.83to 7.0). Whereas, The basal level of Fibrillin-3 at 14 days post insemination was 3.3ng/ml. The range is from 2.76,395 to 3.29 ng/ml. There was no significant difference in the mean of Fibrillin -3hormone between pregnant and non-pregnant women through the phases of menstrual cycle or between the two groups, but there is a significant value between fibrillin-3 post insemination and E2 day 2 and day 13 P<0.05. Conclusion: The present study concluded that Fibrillin -3 hormone has no role in pregnancy, but it may have a main relationship with estrogens reaction in folliculogenesis, oocyte maturation and follicular contents. Therefore, this hormone may be one of the new hormones that interfere with fertilization capacity.
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.