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
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
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.
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.
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.
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.
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.
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
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.
Background: it is important to study a history of the science to appreciate the past as a motive for hard work in the present for better future. In the other hand, the factors affecting pregnancy rate of frozen embryos are yet to be clarified at different embryonic developmental stages. Objectives: to record a brief history of embryo cryopreservation in Iraq and to study many clinical and embryological factors that might affect the pregnancy rate. Subjects, materials and Method: a cross-sectional study of many clinical and em-bryological variable, where120 frozen-thawed embryo transfer cycles enrolled in the study at the period from January 2017 till July 2018. The variables were sta-tistically analyzed first by one factor analysis comparing between pregnant and non-pregnant cases; then by multivariate logistic regression analysis to illustrate main factors without a possible confounder effect. Results: In one factor analysis, the following variables showed a significantly high effect on pregnancy rate (P value <0.001) which are women’s age, women’s weight, number of retrieved oocytes, mature oocytes and good quality embryos, developmental stage, E2 level, and endometrial thickness. While in multivariate logistic regression analysis the women’s weight, the number of retrieved oocytes and good quality embryos were the main factors. Conclusions: it is judiciously to consider these factors while managing infertile couples with embryo cryopreservation programs especially the modifiable fac-tors.
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.
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.
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).
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
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.
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.