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.
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.
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.
Background: Sperm cells are well equipped with a powerful defense system of antioxidants, but an imbalance between the production of reactive oxygen species (ROS) and the available antioxidant-defenses result in oxidative stress. Therefore, antioxidants are supplemented extracellularly under in vitro conditions. Antioxidants are the main defense factors against oxidative stress induced by free radicals. Vitamin E is believed to be the primary component of the antioxidant system of the spermatozoa and is one of the major membrane protectants against ROS and lipid peroxidation (LPO) attack. It appears to be the first line of defense against the peroxidation of polyunsaturated fatty acids (PUFAs) contained in the cellular and sub-cellular membrane phospholipids because of its lipid solubility. Objective: The aim of the current study to evaluate effects of supplementing semen extender with vitamin E, at various concentrations (2, 6 and 12 IU/ml) on sperm motility, survival and morphology on the releases of free radicals and antioxidant enzymes within semen. Patients ,Materials and Methods: Twenty two patients were involved in the present study semen samples were obtained, and sperm count was assessed. The antioxidant (vitamin E) was formulated to be tested at three different levels as follow; 2, 6 and 12 IU/ml; Therefore, control and 3 antioxidant-containing extenders were prepared for semen dilution. The sample was divided into 4 aliquots. One volume of semen was added to 5 equal volumes of the designed extender. Extended semen samples (37˚C) were gradually cooled to 4˚C in the refrigerator and stored for 48 hours. A semen sample was taken out after 48h of storage, then warmed to 37˚C and checked for progressive motility, viability and abnormality. Sperm viability was assessed by Eosin Y. (0.5%)—Nigrosin (0.1%) staining mixture. A total of 200 sperm were assessed under oil immersion with a high-resolution (X100) objective. Sperm morphology was assessed by Hematoxylin-Eosin satin, at least 200 sperm were scored on randomly chosen field, under oil immersion with a high-resolution (X100) objective. Results: The highest post-thaw motility and sperm survival (44% and 51.7%) 48 h after storage at 4˚C (Table 1) were attained (P < 0.01) in the basal diluents containing 6 IU vitamin E. Likewise, the correspondent percentages of live spermatozoa were 59%. Contrariwise, percentage of sperm abnormalities for the previous treatment was 25% significantly (P < 0.05) decrease compares with basal diluents containing 2, 12 IU vitamin E. The lowest (P ≥ 0.05) survival (23.5%, Table 1) was found in the basal diluents containing 12 IU vitamin. Beyond 6 IU vitamin E, there appears to exert adverse effects on sperm viability and survival. Conclusions: For the best protection against the increased free radicals production during chilling-preservation of human semen, extenders must contain 6 IU vitamin E per ml. The intensive production of free radicals in seminal plasma was counteracted by the inclusion of vitamin E at the above mentioned concentrations.
Background: Obesity has been defined as abnormal or excessive fat accumulation that may impair health while the Body mass index (BMI) is a simple index of the weight-to-height ratio that is commonly used in classifying overweight and obesity in adult populations. Many semen parameters alteration can be ascribed to obesity like decreased sperm concentration, abnormal morphology, compromised chromatin integrity and abnormal motility. Many evidences showed an adverse effect of excess body fat on spermatogenesis. Methods: A total sample size composed of (70)infertile men were grouped into three groups according to published BMI ranges as follows: normal weight, overweight and obese.A single semen sample was collected for each infertile men.Heightin (m) and weight in (kg) were recorded on day of semen collection.BMI was calculated as BMI= Kg\m2. Sperm parameters evaluated according to World Health Organization guidelines.Quantitation of the IL-6 in seminal plasma were accomplished using commercially available ELISA kit. Results: In the current study, a positive correlation was found between body mass index and IL-6, on the contrary, a significant negative correlation was found among IL-6 level and progressive sperm motility and sperm concentration as well. Conclusion: There is an increase of IL-6 from the adipose tissue with increasing BMI. The IL-6 cause reduction of the sperm motility by increasing the level of ROS. The obesity reduces the spermatogenesis by androgenic deficiency results in low sperm concentration and motilitywhich it may be the main reason of their infertility.
The purpose of the study was to determine if age and body mass index in infertile men were associated with stress levels as determined by salivary stress biomarkers (cortisol and alpha-amylase). A cross-sectional study comprising 100 males with an infertility challenge was divided into 25 normozoospermia, 26 oligozoospermia, 21 asthenozoospermia, and 28 oligoasthenoteratozoospermia. Salivary cortisol and alpha-amylase were examined using the ELISA technique . The average age of the patient was 30.85±7.03, and their average BMI was 27.604±5. 7894. The infertile duration mean was 4.7896±3. 85932. The average amylase content in the saliva was 127.3721±40.0952, while the average cortisol content was 72.67±49.09. There are no discernible variations between the comparison mean of BMI and the duration of infertility. In the current investigation, there was no discernible relationship between SAA and age, BMI, or the length of infertility. Saliva cortisol and BMI in the oligoasthenoteratozoospermia group showed a substantial negative correlation (r = 0.483 & p = 0.009), although no significant link was seen with age, BMI, or length of infertility in the other study groups. There was no significant positive or negative correlation found between SAA level and the age and BMI of the four study groups, whereas salivary cortisol showed a substantial negative correlation with BMI in OAT and a non-significant correlation with age.