Inhibition of luteinizing hormone (LH) by gonadotropin releasing hormone (GnRH) antagonist may lead to suboptimal response during ovarian stimulation. In addition, several studies suggest that low level of LH is associated with lowered fertilization and implantation rate and increased early pregnancy loss rate. The aim of this study is to study the effect of adding low dose human chorionic gonadotropin (hCG - 200 IU), as an LH supplement, to recombinant follicle stimulating hormone (rFSH) in a GnRH antagonist cycles in women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment. Sixty-three infertile women undergoing IVF were randomly divided into two groups. One group was stimulated with the conventional stimulation protocol (rFSH alone), while the second group received 200 IU hCG in addition to rFSH in the late follicular phase (hCG + rFSH). Both groups' results including pregnancy rate, total dose of rFSH required, duration of stimulation, endometrial thickness, oocytes and embryos characteristic, serum hormone levels (Testosterone, Estradiol, Progesterone, and LH) and level of epidermal growth factor (EGF) were compared. The results show that pregnancy rate among the group of women who received the low dose hCG was higher than those who did not receive hCG. However, this difference did not reach statistical significance. Furthermore, other cycle outcomes and hormonal values were comparable between the two stimulation protocols.
Non-steroidal anti-inflammatory medications may interfere with ovulation, implantation, and pregnancy; they are generally avoided during reproductive procedures. The risks of using them during IVF must be carefully weighed against the necessity of pain relief during oocyte retrieval. About 64 women (ages 20–42) undergoing ICSI participated in a single-blind, cross- sectional clinical trial at the High Institute for Infertility at Al-Nahrain University from October 2024 to May 2025. During oocyte retrieval, participants were split into two groups at random and given propofol anesthesia and ketorolac analgesia, respectively. They all adhered to a common protocol for ovarian stimulation. Based on their maturity stage (MII, MI, or GV), oocytes were categorized; those that were aberrant or degenerated were not included in ICSI. ELISA was used to measure the levels of cortisol and serum LH both before and after oocyte retrieval. The mean post -operation LH Marker level were 10.41±8.0, 6.40± 7.48 among GA and analgesia groups respectively, this relation was statistically significant (P value =0.04). The mean Post -operation Cortisol Marker level were 201.13±132.34, 159.13± 78.42 among GA and analgesia groups respectively . The mean number of Oocytes: Similar for both groups (10±5.53 for anesthesia and 10.59±5.44 for analgesia). Abnormalities of Oocytes: slightly lower in the group receiving anesthesia (0.56±1.44 vs. 0.69±1.18). The anesthesia group had fewer ruptured oocytes (0.16±0.45 vs. 0.25±0.57). Fertilized embryos: Grade I: The anesthesia group had slightly fewer (2.75±2.63 vs. 3.13±2.46). Grade II: Groups are comparable (1.41±1.58 vs. 1.31±1.53). Grade III: Lower in the group under anesthesia (0.19±0.54 vs. 0.38±0.83). Similar results were obtained from arrested embryos (anesthesia: 0.63±1.1, analgesia: 0.68±1.64). The analgesia group had a significantly higher number of transferred embryos (anesthesia: 1.84±1.08, analgesia: 2.88±1.62; P = 0.004). The analgesia group showed a significant decrease in post-operation LH marker, Cortisol Marker, and higher mean number of transferred embryos compared to the anesthesia group.
Background:Ovulation disorders, usually presents as menstrual disturbance, are the cause of infertility in around 25% of couples who have difficulty to conceive. The drug most commonly prescribed for the induction of ovulation is clomiphene citrate (CC). If the patient does not ovulate after the use of CC, the choice was to add gonadotropins. Gonadotropin therapy is very expensive with significant risks of high order multiple gestations and ovarian hyperstimulation syndrome. Human chorionic gonadotropin hCG can function as a surrogate for LH and oc-cupies LH receptors for more than 24 hours allowing prolonged stimulation also it has longer half-life and greater affinity for the LH receptor. Objective: The purpose of this study was to compare the effectiveness of low-dose human chorionic gonadotropin (hCG) in the late follicular phase to induce ovulation in clomifene citrate resistant patients who had previously failed to ovulate on clomiphene citrate (CC) alone. Subjects,Materials and Methods: A total of 71 patients who attend infertility clinic in the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, where prospectively randomly assigned into two groups. hCG Group received 100 mg dose of CC from day 5 of men-strual cycle for 5 days, then each patient received 200 IU hCG (DICLAIR®) subcutaneous injection daily when the largest follicle mean diameter was 12 mm or larger starting on day twelve of menstrual cycle. Non-hCG group received 150 mg dose of CC from day 5 of menstrual cycle for 5 days, and both groups were monitored with transvaginal ultrasound. Ultrasound measurements of follicle number and growth, ovulation, endometrial thickness and pattern were recorded and compared between the two groups. Student t test and fisher exact test were used for statistical comparison between the two groups. Results: The low-dose hCG group had significantly higher percentage of ovula-tory cycles (80% vs 44.4% P value 0.006). There was no significant difference in the endometrial thickness between the groups but it appears higher in hCG group (10.55±1.82 vs. 9.62 ± 1.87 in non-hCG group, p value 0.056). Non significant but higher incidence of echogenic (luteinized) endometrium 48 hour post ovula-tion in hCG group (36/40 (90%) vs. 20/25 (80%) for non-hCG group p-value = 0.288). Conclusions: The use of low-dose hCG after CC in the late follicular phase re-sults in continued follicle growth, higher ovulation rate. This treatment offers an efficient and cost-effective alternative before gonadotropin therapy for this type of patients.
The crosstalk between a receptive endometrium and a functional blastocyst during human embryo implantation is crucial for conception. Because platelets rich plasma (PRP) with concentrated platelets 4-5 times higher than normal, when release of granules containing growth factors including VEGF, TGF, PDGF, IGF, and EGF, these factors involved for sub endometrial angiogenesis and endometrial receptivity. 44 women under the age of 40 were given antagonist ovarian stimulation treatments, the oocytes were harvested utilizing 2-D power doppler ultrasound guidance, then ICSI done for them. On the hCG day all had an intrauterine autologous PRP infusion. Power Doppler is utilized to measure endometrial thickness (EnT), pulsatility index (PI), and resistance index (RI) of sub endometrial arteries, as well as serum levels of VEGF and EGF were measured on the hCG day and ovum pickup (OPU). The ultrasound findings, on the hCG day compared to those of OPU in all women with intrauterine infused autologous PRP were highly significant (HS). The mean EnT, RI, PI, VEGF and EGF at OPU day was lower than that on hCG day in a HS manner, with (p < 0.001), (p < 0.001), (p=0.047), (p < 0.001), and (p<0.001) respectively. These characteristics were shown to be significantly and independently associated to intrauterine PRP infusion. After PRP injection, both growth factors serum levels (VEGF and EGF) increased, and ultrasonography sub-endometrial metrics such as EnT, RI, and PI changed as the thickness grew while vascular resistance decreased, and all considered as predictors of endometrial receptivity.
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
Despite many improvements in Assisted reproductive techniques during the last three decades, clinical pregnancy and live-birth rates still nearly 30–40% and 20–30%, respectively. To enhance endometrial receptivity many attempts done one of the endometrial scratching could provoke delay in endometrial maturation reforms asynchrony between endometrium and the concepts and indeed induce wound healing by producing a considerable raise in the local secretion of pro-inflammatory cytokines and growth factors like Heparin Binding Epidermal Growth Factor (HBEGF) and its receptors has role in mediate the receptivity, maturation, and decidualization of endometrium and facilitates the implantation and development of both placenta and embryo. All women were undergone IUI procedure. They divided into two groups. Intervention group who did endometrial scratching by using pipelle catheter in luteal phase of preceding cycle and another blood sample aspirated before scratch. Control group include women without intervention of endometrial scratch. Both groups underwent controlled ovarian stimulation, triggering of ovulation and another blood sample was aspirated at day of IUI to measure level of HBEGF. 14 days later with luteal support, BHCG test underwent in blood to check pregnancy. Higher pregnancy rate among patients with scratch group (5/20) 25% than non-interventional group (3/25)12%. These result showed that HBEGF level in scratch group (386.1±124.7) is significantly higher than control group (279±189.3) as P-value was (0.035). Moreover, the HBEGF level at day of IUI (455.67±157.34) and in pregnant (563.9±163.66) is significantly higher than before scratch in scratch group (279.0±124.67), (406.26±138.89), as P-values were (0.00007, 0.049) respectively. Intentional local damage to the endometrium is a cost–effective procedure can be used to enhance the uterine receptivity that may help to overcome implantation failure problem.
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.
Despite developments in assisted reproductive technology, there is immaterial progress in the implantation and pregnancy rates. Intrauterine infusion (IUIF) of autologous platelet-rich plasma (PRP) might renew implantation rates through its paracrine properties by progression cytokines and growth factors which favor implantation. Here we determine whether the IUIF of autologous PRP had a role in pregnancy outcome through its outcome on epidermal growth factor and endometrial thickness. An overall of 43 patients where prospectively randomly dispersed into two groups subjected to a superovulation program using Letrozole® tablet orally 2.5 mg twice daily 12 hours apart from day 2 for 5 days for one cycle. 20 women were considered as control receiving the conventional intrauterine insemination (IUI) management while 23 of them were given PRP by IUIF on the day of human chorionic gonadotrophin injection. The IUI was done for both groups 36-48 hours after confirming ovulation. The blood samples were collected from both groups on the day of IUI for the valuation of epidermal growth factor and an ultrasound was done on the day of human chorionic gonadotrophin injection and day of IUI for assessment of endometrial thickness. The mean endometrial thickness in the PRP group at the day of IUI was significantly thicker than that of the control group and the difference in percentage change of endometrial thickness between PRP group and controls significantly higher in PRP group. The mean epidermal growth factor and the pregnancy rate were significantly superior in the PRP group than that of controls. In conclusion, autologous PRP IUIF was well-tolerated and resulted in a significant expansion in endometrial thickness, epidermal growth factor Level and, subsequent pregnancy rate in an infertile woman undergoing IUI.
Female infertility is caused by ovulation issues, tubal disorders, uterine abnormalities, endometriosis, and peritoneal factors. For years, intrauterine insemination has been used to treat infertility alone or with ovarian stimulation. Obtaining adhesion ligands and losing inhibitory components are necessary for pregnancy. LIF promotes and differentiates proliferation and differentiation in embryonic, hematopoietic, and endothelial cells. LIF also controls prostaglandins (PGs), which mediate implantation and decidualization. The measuring hormone profile on cycle day 2 before control ovarian stimulation including FSH, LH, E2, Prolactin, TSH, and AMH. Serum LH, E2. The progesterone level was estimated on the day of triggering. The effect of LIF gene genotypes on hormone level. The hormone levels of 100 intrauterine inseminated women were measured on the second day of the cycle and the day of ovulation. DNA extraction for LIF gene variation detection utilizing sequencing and HRM. The current study found no significant differences between pregnant and non-pregnant groups for hormone levels, genotype, and genetic effect on female hormone levels with (P ≥0.05). The LIF gene did not affect hormone levels in infertile females may be due to the tiny samples or the complex signaling chain that happens during implantation.
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
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.
Poor ovarian responders (PORs) are characterized by decreased the response of the ovaries to stimulation. Nowadays, using soft protocol of low doses of Gn with/without oral compounds such as letrozole was accepted as a good option. The endometrium receptive parameters include sub endometrial resistance, pulsatility index and endometrial thickness. Eighty infertile POR women divided to two equal groups and undergoing IVF/ICSI antagonist protocols. One group underwent soft protocol using letrozole 2.5mg twice daily for 5 days starting from cycle day 2-3 overlapped low-dose Gn (225 IU) from cycle day 4-5. Other group received high dose Gn only (450 IU) from cycle day 2-3. Both groups continued stimulation cycle until the day of trigger using the flexible antagonist protocol. Estimation of values of sub endometrial RI and PI with endometrial thickness by 2 Color Doppler ultrasound was performed at day of trigger, OCP and ET. There was no significant difference for endometrial thickness at day of trigger, OCP and ET. There was significant difference in means of sub endometrial RI and PI at day of trigger, OCP and ET (P>0.05). This study showed evidence in favor of soft protocol for POR regarding pregnancy rates in spite of the statically difference in means of sub endometrial RI and PI associated with much less cost effectiveness.
Granulocyte colony-stimulating factor (G-CSF), traditionally utilized in hematopoietic disorders, has garnered attention in reproductive medicine for its potential to enhance endometrial receptivity and improve pregnancy outcomes in assisted reproductive technology (ART). This study aimed to compare the effect of G-CSF administration via intrauterine infusion versus subcutaneous injection on pregnancy outcomes in infertile women undergoing IUI, 75 infertile women aged 18–40 years were enrolled and randomly assigned into three groups (n=25 each): Group 1 received 300 µg G-CSF via intrauterine infusion, Group 2 received 300 µg G-CSF via subcutaneous injection, and Group 3 served as the control with no G-CSF treatment. All participants underwent controlled ovarian stimulation followed by IUI. Endometrial thickness and sub-endometrial blood flow indices (PI, RI, V1/V2) were evaluated using Doppler ultrasound. Serum levels of TNF-α, IL-10, and VEGF were measured before and after G-CSF administration. Clinical pregnancy was confirmed by β-hCG testing two weeks after IUI and confirmation by transvaginal ultrasound. Both G-CSF intervention groups demonstrated significant improvements in sub-endometrial blood flow parameters compared to controls, with intrauterine infusion showing the most favorable Doppler indices (p<0.05). Additionally, intrauterine G-CSF administration resulted in upregulation of VEGF and IL-10 and a reduction in TNF-α levels, though differences did not reach statistical significance. Clinical pregnancy rates were higher in the intrauterine group compared to the subcutaneous and control groups, though differences did not reach statistical significance. Intrauterine infusion of G-CSF appears to offer superior benefits in enhancing endometrial receptivity, modulating inflammatory and angiogenic biomarkers, and improving pregnancy outcomes in IUI cycles compared to subcutaneous injection or no treatment. Further large-scale studies are warranted to validate these findings.
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.