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).
Background: Citrullus colocynthis(CC) is an herbal medicine used in the treatment of a wide range of diseases. Recently, the effect of this plant on the reproductive system has been studied.However, its role in vitro still unclear. Objective: The present study was designed to investigate the possibility of using CC extract for in vitro sperm direct activation technique, in vitro fertilization(IVF) and early embryonic development using the mice as a model for mammals. Materials and Methods: Citrullus colocynthis extract (0.05mg/ml culture medium) was used for in vitro direct sperm activation technique. The same medium was used for oocytes insemination and for culturing the embryos after 24-48 hours of insemination. The oocytes were collected from superovulated female mice and divided into two groups: first group, 343 oocytes were inseminated and cultured in CC- free Ham's F-12 medium (the control group). The second group, 345 oocytes were inseminated and cultured in 10% CC - Ham's F-12 medium (treated group). Each 4 oocytes were inseminated with the same sperm concentration (1-2 x 10^5 sperm/IVF well). The fertilization rate was recorded after 24 hours of insemination, while embryonic development rate was recorded after 24 and 48 hours of insemination. Results: In vitro activation of epididymal sperms with 10%CC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a highly significant (P<0.004) increase in FR of the treated group (61.5%) compared to control group (50.4%) after 24 hours of insemination. Embryonic developmental rate was significantly increased after 24 and 48 hours of insemination in treated group compared to control group. Conclusion: It concluded that adding the 10%CC to the culture medium of the epididymal sperm and in vitro inseminated lead to improvements in certain sperm function parameters and sustain the FR and early embryonic development rate.
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.
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.
Cigarette smoke contains a substance which is a toxic mutagenic chemical, and carcinogenic compounds, which have a negative effect on male fertility. Smoking cigarettes is associated with noticeably higher levels of the oxidative stress-causing reactive oxygen species in the seminal fluid. Because of this, smoking may cause changes to the sperm plasma membrane and a significant amount of deoxyribonucleic acid fragmentation. To study the influence of smoking on sperm chromatin immaturity and ICSI outcomes (fertilization rate, embryo grading and pregnancy rate). At the laboratories of the High Institute of Infertility Diagnosis and Assisted Reproductive Technologies and after 1-7 days of sexual abstinence, 92 males’ sperm samples were taken. After liquefaction, macro-and microscopic studies were carried out, criteria applied by the WHO 2021. Masturbation was used to obtain the sample, which was then placed in sterile containers. The relationship between the smoking and chromatin immaturity and ICSI outcomes have been evaluated. A significant positive correlation between smoking status and sperm chromatin immaturity. Furthermore, there was a negative non-significant correlation between smoking status and fertilization rate. Weak and non-significant correlations between smoking status and embryo grade 2 as well as embryo grade 3. There was a positive association between smoking status and embryo grade 1. The sperm chromatin integrity was significantly lower in the non-smoker group compared to the smoker group.
Background: Many studies has been found that the pentoxifylline (PX) and L-carnitine (LC) as motility stimulants showed a positive effect on the activation of sperm in vitro and improve the forward movement after cryopreservation. Objectives: The present study aim to investigate the possibility of using the motility stimulants(i.ePX,LC and mixture of PX with LC medium) for in vitro sperm activation after cryopreservation of epididymal sperms and their effects in vitro on fertilization rate (FR) and early embryonic development (ED) following in vitro fertilization (IVF) process as a model for obstructive azoospermia in men. Methods: Fifty mature male and Fifty mature female mice, 8-12 weeks old were used. Epididymal sperms were obtained from caudal region and in vitro direct activation technique was done after cryopreservation by using four media namely; PX alone, LC alone, a mixture of PX with LC and Ham's F-12 (as control media). The four media were used for IVF of mature ova and FR with ED were recorded. Results: In vitro activation of epididymal sperms with PX, LC and mixture of PX with LC has shown positive effect on sperm concentration, sperm motility, and grade activity of progressive forward movement after cryopreservation. There was a significant (P<0.05) increase of FR in treated media compared with the control medium after 24 hours of insemination. The study showed that the quality and quantity of embryos. Conclusion: It is concluded from the results of the present study that adding PX, LC and mixture of PX with LC medium to the culture medium of cryopreserved epididymal sperms and IVF leads to an improvement in certain sperm function parameters, supports the FR and early ED rate.
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.
The aim of any given ICSI cycle is to achieve normal response and avoid poor or hyper response. A part from ovarian hyper stimulation syndrome (OHSS), pregnancy rate in hyper responder females usually low which could be related to the adverse effect of hyper estrogenic state on oocytes, embryos quality and uterine endometrium. To evaluate the effect of hyper response on ICSI outcome represented by oocytes, embryos quality, fertilization and pregnancy rates. Study showed that total number of retrieved oocytes, mature, immature oocytes, total embryos, good quality embryos and blastocyst count were significantly higher in hyper followed by normal and poor responders respectively. Fertilization rate was significantly higher in both hyper and normal responders than poor ones, while pregnancy rate was significantly higher in hyper responders 47.6% followed by normal 40.7% and then poor ones 16.7%. Conclusions: Irrespective of abnormal hormonal milieu, excessive ovarian response and higher number of produced oocytes improve ICSI outcome in hyper responder females by increasing the chance of getting higher number of embryos, good quality embryos and even blastocyst numbers which in turn increase the chance of pregnancy.
Background: Diabetes has been found to induce various subtle molecular changes important for sperm quality and function: The mammalian spermatozoa are rich in polyunsaturated fatty acids, making them particularly sensitive to the deleterious effects of lipid peroxidation, which may result in irreversible loss of motility and a low level of fertilizing capacity. An antioxidant, glutathione (GSH) a polypeptide is important in biological oxidation-reduction reactions and improves sperm motility. Objective: The study was aimed to investigate the effects of GSH injection on healthy and diabetic male mice fertilization potential in vitro following in vitro fertilization (IVF). Materials and Methods: Inducing diabetes in mice through intra peritoneal alloxan injection. The treatment with GSH divided into two periods 10 and 20 days, studying the in vitro fertilization outcomes. Results: Glutathione daily IV injected in a period of 10 to 20 days for the diabetic induced male mice significantly increase the sperm concentration and motility and decrease the abnormal and dead sperms percentages in treated group compared to untreated group. The Fertilization rate was significantly (P<0.05) increased in mice treated with GSH compared to untreated one. Conclusion: Glutathione injections may decrease the oxidative stress caused by diabetes and enhance the in vitro fertilization potentials in male mice.
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.
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.
Stem cell factor (SCF) is one of the first growth factors derived from granulosa cells in the ovarian follicle plays a critical role in hematopoiesis and the generation of melanocytes and germ cells. SCF also serves an important role in the development of the interstitial cells of Cajal in the intestine and the learning functions in the hippocampal region of the brain. This study aimed to investigate the levels of SCF in follicular fluid (FF) and if it can be used as a potential marker for predicting oocyte, embryo quality, and pregnancy rate. A total of 44 infertile couples involved in the study. All of them underwent controlled ovarian hyperstimulation for the ICSI cycle. Antagonist protocol was used as an ovulation induction protocol. Patients were divided into two groups according to ICSI outcomes as pregnant (23 women) and non-pregnant (21 women) groups. A comparison was done in both groups based on the oocyte, embryo quality, and other ICSI cycle characteristics, and follicular fluid stem cell factor level. SCF level was measured by using an enzyme-linked immunosorbent assay. Our study showed lack of association between patients' characteristics which included (age, BMI, infertility duration, type of infertility, and causes of infertility, basal hormonal, stimulation characteristics levels), FF SCF level and the oocyte characteristics comparison (total number retrieved oocyte, germinal vesicle oocyte (GV), MI oocyte and MII oocytes, and the number of injected oocytes, fertilization rate, and embryo characteristics) between pregnant and non-pregnant groups. The present study showed that Mean follicular fluid SCF was significantly lower in pregnant women in comparison with that of non-pregnant women, 135.92±38.96 pg/ml versus 185.15±63.58 pg/ml, respectively (P=0.003). Raised SCF level in FF harms ICSI outcome.
Background: Glycyrrhiza glabra(Gg) is a herbal medicine used in the treatment of different syndromes. Recently, the effect of this plant on the reproductive system has been studied. However, its role in in vitro still not clear. Objectives: The present study aims to investigate the possibility of using Glycyrrhiza glabra extract for in vitro sperm direct activation, and its effect on in vivo fertilization rate (FR) and early embryonic development (ED) following intra-peritoneal insemination (IPI). The mice were used as an experimental model for mammals. Materials and Methods: Glycyrrhiza glabra extract (2 mg/ml culture medium) was used for in vitro direct sperm activation technique. The female mice were divided into two groups: spontaneously ovulated (SOM) and superovulated groups (SuOM). Each group was divided into two subgroups, the first: the IPI was accomplished by epididymal sperm activated in vitro by adding 20%Gg to the culture medium. The second subgroup: the IPI is accomplished without adding Gg to the culture medium. Results: In vitro activation of epididymal sperms with 20% Gg has shown positive effects on sperm concentration, sperm motility, and grade activity of progressive forward movement. There was a significant (P<0.05) increase in FR with adding 20%Gg compared with that free of Gg of spontaneously ovulated groups after 24 hours of insemination. The FR in the right oviduct showed highly significant (P<0.001) increment compared to the left. Embryonic development rate significantly ( (P<0.05) increased after 24 and 48 hours of insemination in grouping with adding Gg. The study showed that the quantity and quality of embryos generated from IPI with adding Gg were higher than that of groups inseminated without adding Gg. Conclusion: It is concluded from the results of the present study that adding the 20% Gg to the culture medium of the epididymal sperm and IPI leads to an improvement in certain sperm function parameters and supports the FR in SOM and SuOM with an increase in the embryonic development rate. These data can be utilized for artificial insemination programs.
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.
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.