Stress In Follicular Fluid Research Articles

The Impact of Follicular Fluid Oxidative Stress Levels on the Outcomes of Assisted Reproductive Therapy.

Oocyte quality is a pivotal determinant of assisted reproductive outcomes. The quality of oocytes is intricately linked to their developmental microenvironment, particularly the levels of oxidative stress within the follicular fluid. Oxidative stress levels in follicular fluid may have a substantial influence on oocyte health, thereby impacting the outcomes of ART procedures. This review meticulously explores the intricate relationship between oxidative stress in follicular fluid and ART outcomes. Furthermore, it delves into strategies aimed at ameliorating the oxidative stress status of follicular fluid, with the overarching goal of enhancing the overall efficacy of ART. This research endeavors to establish a robust foundation and provide valuable guidance for clinical treatment approaches, particularly in the context of infertile women, including those of advanced maternal age.

Oxidative Stress in Follicular Fluid of Low Responder and Hyporesponse Patients Undergoing the IVF Program at Halim Fertility Center, IVF Clinic, Division of Reproductive Endocrinology, Faculty of Medicine, USU Medan

Oxidative Stress in Follicular Fluid of Low Responder and Hyporesponse Patients Undergoing the IVF Program at Halim Fertility Center, IVF Clinic, Division of Reproductive Endocrinology, Faculty of Medicine, USU Medan

Assessment of oxidative stress in follicular fluid of infertile women undergoing IVF procedure

Oxidative stress (OS) plays an important role in biological processes of reproduction, and yet there is no accurate indicator to measure it. The objective of the study was to correlate the effect of ROS (reactive oxygen species) on embryo quality and pregnancy outcome. This was achieved by measuring levels of ROS present in granulosa cells and follicular fluid (FF) of women undergoing IVF (in vitro fertilization). ROS in FF is measured using thiobarbituric acid reactive substances assay kit (Abcam) and granulosa cells were measured with the help of flow cytometry. The results showed that high levels of FF malondialdehyde were found in nonpregnant women as compared to pregnant group and were found to be significant (P 0.05) but had an impact on the quality of embryos (P

Assessment of oxidative stress in follicular fluid of infertile women undergoing IVF procedure

Assessment of oxidative stress in follicular fluid of infertile women undergoing IVF procedure

Role of oxidative stress in follicular fluid on embryos of patients undergoing assisted reproductive technology treatment.

Oxidative stress (OS) is defined as an imbalance between oxidants and antioxidants in favor of the oxidants, and the disruption of redox signaling leads to molecular damages. This study was conducted to clarify the effect of OS in individual follicular fluid (FF) on oocyte fertilization and embryonic division. A total of 124 patients who underwent assisted reproductive technology treatment in our hospital underwent intracytoplasmic sperm injection and 211 FF were collected. The ova were fertilized, and embryos were individually cultured until cleavage stage or blastocyst stage and classified according to the Veeck classification system or Gardner classification system. The follicular fluid corresponding to the ovum was analyzed by measuring the OS marker, the reactive oxygen metabolites (d-ROM test), and the antioxidant marker (biological antioxidant potential marker [BAP] test), and the relation between these markers and clinical parameters was analyzed. The value of d-ROM was correlated with the proper fertilization status and formation of good quality cleavage embryo, whereas the elevated value of BAP was observed in better embryonic development group. Oxidative stress index, defined as d-ROM/BAP × 100 clearly indicated that lower oxidative stress index was associated with better fertilization status and embryo development. Our results clearly indicate that the balance between OS and antioxidant capacity in FF at the time of oocyte retrieval is possibly important in the processes of fertilization and embryo division; thus, we propose that oxidative status and balance in FF might be used as novel biomarkers in assisted reproductive technology.

The role of stimulation protocol, number of oocytes retrieved with respect to follicular fluid oxidative stress and IVF outcome

The study was conducted on a population of 203 women attending the IVF clinic, Ahmedabad, India, to explore the role of stimulation protocol and the number of oocytes retrieved on oxidative stress in follicular fluid and IVF outcome. Follicular fluid was collected during ovum pick-up to determine the oxidative stress markers: superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), total thiols (TT), l-ascorbic acid (AA), total protein, glutathione S-transferase (GST) and glutathione reductase (GRD). The average number of oocytes retrieved was significantly higher in the women receiving a long GnRH agonist protocol compared to GnRH antagonist protocol while the percentage of women with positive IVF outcome was lower in the long agonist protocol. The level of total thiols was significantly lower in the group of women administered a short agonist protocol. The proportion of ETs carried out and positive IVF outcome were higher following retrieval of an intermediate number of oocytes (6–10 oocytes) compared to a lower (0–5 oocytes) and higher (>10 oocytes) number. Mean glutathione reductase (GRD) activity in follicular fluid was significantly elevated in the intermediate (6–10) and higher (>10) oocyte retrieval groups compared to the lower oocyte retrieval group. Positive IVF outcomes were highest when oocyte retrieval was in the range of 6–10 oocytes, and the level of MDA was lower (1.76 ± 0.13 nmol/ml) as compared to 0–5 and >10 oocytes retrieved groups.

Determination of oxidative stress balance in follicular fluid

AbstractBackground:The role of oxidative stress in female reproduction is an area that needs more investigation. This study aims to assess the oxidative stress levels in follicular fluid (FF) samples of women undergoing in vitro fertilization (IVF) and to relate them to different diagnoses.Methods:A total of 199 woman were enrolled; 37 of them were diagnosed with polycystic ovary syndrome (PCOS), 71 with endometriosis and 41 with tubal factor infertility (TFI) and 50 of the patients were used as a control group. A sample of FF was collected from each patient at an oocyte retrieval day and analyzed for oxidative stress by measuring a class of reactive oxygen metabolites (ROMs) (dROMs test) and also analyzed for the plasma antioxidant test (PAT).Results:The data analyzed in FF were related to different diagnoses. Groups were not significantly different in age and body mass index (BMI), except for the PCOS group. There was a significant difference between dROMs and PAT levels in FF of patients vs. control group. The same finding was seen when the dROM/PAT ratio was used.Conclusions:We conclude that the evaluation of oxidative stress in FF needs more investigation with regard to markers in the follicular microenvironment.

Antioxidants reduce oxidative stress in follicular fluid of aged women undergoing IVF

BackgroundThe status characterized by the imbalance between pro-oxidants and antioxidants molecules, defined as oxidative stress, has been suggested to be involved in the pathogenesis of subfertility in females. This study aims to evaluate the impact of a complete micronutrients supplementation on oxidative stress levels in follicular microenvironment as well as on in vitro fertilization (IVF) outcome.MethodsThis preliminary study was conducted between January 2014 and July 2015 at the Siena University Hospital Infertility Clinic. Serum and follicular fluid were collected from infertile women aged > 39 years who underwent two in vitro fertilization cycles: in the first cycle they were treated with GnRH-antagonist protocol and gonadotropins for controlled ovarian hyperstimulation, whereas in the second cycle ovarian stimulation protocol was associated to micronutrients supplementation, starting three months earlier. Protein oxidation levels and total antioxidant capacity in serum and in follicular fluid were evaluated in IVF cycles with or without micronutrients supplementation. Differences in IVF outcome parameters were statistically evaluated.ResultsTwo-dimensional electrophoresis analyses demonstrated that when patients assumed micronutrients before IVF cycles, follicular fluid and serum proteins were protected from oxidative damage. Comparable results were obtained when total antioxidant capacity was measured. Moreover, the mean number of good quality oocytes retrieved when patients received micronutrients supplementation was significantly increased.ConclusionThe additional treatment with micronutrients, starting three months before IVF cycles, protects the follicular microenvironment from oxidative stress, thus increasing the number of good quality oocytes recovered at the pick up.

Oxidative stress in follicular fluid of young women with low response compared with fertile oocyte donors

The aim of this study was to determine the concentrations of oxidative stress markers, antioxidant enzymes and cytokines in the follicular fluid of young women with low response in ovarian stimulation cycles compared with high responders and fertile oocyte donors of the same age, to assess the impact of oxidative stress on ovarian reserve. The activity of follicular fluid antioxidant enzymes glutathione transferase, glutathione reductase and glutathione peroxidase was significantly lower in young women with reduced ovarian reserve compared with that in high responders and oocyte donors. Follicular fluid concentrations of oxidative stress marker malondialdehyde combined with 4-hydroxyalkenals and nitric oxide were higher in low responders than in high responders and oocyte donors. Significant differences between low responders and donors in concentrations of IL-2, IL-6, IL-8 and vascular endothelial growth factor were observed, with higher concentrations in low responders. However, IL-10 concentration was lower in low responders than in high responders and donors. No significant differences were found in follicular fluid concentrations of tumour necrosis factor alpha between the three groups. These results demonstrate that different concentrations of oxidative stress markers, oxidant enzymes and cytokines in low responders compared with high responders and oocyte donors may negatively impact ovarian response.

Markers of oxidative stress in follicular fluid of women with endometriosis and tubal infertility undergoing IVF

Oxidative stress and trace elements in the oocytes environment is explored in endometriosis and impact on in vitro fertilization (IVF) outcome assessed. Follicular fluid was aspirated at the time of oocyte retrieval from endometriosis (n=200) and tubal infertility (n=140) and the analytes measured using spectroscopy and HPLC. Increased concentration of reactive oxygen species (ROS), nitric oxide (NO), lipid peroxidation (LPO), iron, lead, cadmium and reduced levels of total antioxidant capacity (TAC), superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR), vitamins A, C, E, copper, zinc and selenium was observed compared to tubal infertility. Increased ROS and NO in endometriosis and tubal infertility associated with poor oocytes and embryo quality. Increased levels of ROS, NO, LPO, cadmium and lead were observed in women who did not become pregnant compared to women who did. Intrafollicular zinc levels were higher in women with endometriosis who subsequently became pregnant following IVF.

Obesity is not a descriptive factor for oxidative stress in follicular fluid of in-vitro fertilization patients

Obesity is not a descriptive factor for oxidative stress in follicular fluid of in-vitro fertilization patients

Analysis of follicular fluid and serum markers of oxidative stress in women with infertility related to endometriosis

To study the levels of four markers of oxidative stress in follicular fluid (FF) and plasma of patients with infertility related to endometriosis and controls. Experimental study. University-affiliated hospital and infertility center. Ninety-one infertile women were included in the study (23 infertile women with endometriosis and 68 controls including infertile women due to tubal factor, male factor, or healthy egg donors). Blood was obtained at the time of egg retrieval, and FF from the mature follicles of each ovary was centrifuged and frozen until analysis. Vitamin C and E, malondialdehyde, and superoxide dismutase concentrations in plasma and follicular fluid. Women with endometriosis showed a lower vitamin C concentration in FF (12.7 ± 5.9 vs. 9.7 ± 6.9 μg/mL) and lower superoxide dismutase concentration in plasma (0.9 ± 1.4 vs. 0.5 ± 0.7 U/mL) compared with controls. Vitamin E plasma levels were significantly higher in women with endometriosis (8.1 ± 3.8 vs. 5.2 ± 3.2 μg/mL). A nonsignificant trend toward a lower plasma concentration of malondialdehyde was found in women with endometriosis. These findings suggest a lower antioxidant capacity in infertile women with endometriosis. Although a certain level of reactive oxygen species is required under physiological conditions, an altered balance between pro-oxidant and antioxidant activities may have an impact on folliculogenesis and adequate embryo development.

Oxidative stress in follicular fluid of patients submitted to controlled ovarian stimulation cycles

OBJECTIVE: Oxidative stress is an important cause of female infertility. During follicular development, oxidative stress can induce oocyte degeneration and apoptosis, modifying the meiotic spindle and causing lipid peroxidation, loss of membrane selectivity, inactivation of cytoplasmic enzymes, and DNA damage. The aim of this study was to verify relations between concentrations of lipid peroxides (TBARS) and activity of the antioxidant enzymes glutathione peroxidase (GPx) and catalase (CAT) with the occurrence of pregnancy, hormonal stimulation protocols and presence of a female infertility factor.DESIGN: Prospective study.MATERIALS AND METHODS: Follicular fluid from 146 patients submitted to ICSI was included. Groups were divided according to pregnancy, hormonal stimulation protocol (with or without LH), and presence or not of at least one female infertility factor. Follicular fluids were collected after oocyte retrieval, and TBARS levels and CAT and GPx activity analyzed by spectrophotometry. Student's T-test or ANOVA were used, and logistic models were calculated using embryo quality on day 3, presence of female infertility, and TBARS levels and CAT and GPx activity as independent variables, occurrence of pregnancy as a binary dependent variable.RESULTS: In the non-pregnant group an increase in GPx activity (p=0,04) was observed. In the other groups no differences were observed. Logistic regression produced a model which best predicted pregnancy including presence of female infertility factor, embryo quality on day 3 and GPx activity. Odds-ratio for GPx activity was 0.990 (increasing values of GPx activity decrease the odds of occurrence of pregnancy).CONCLUSIONS: We may conclude that (i) higher GPx activity is associated to negative outcomes in ICSI cycles, (ii) inclusion of LH in ovarian stimulation protocols does not alter TBARS concentrations nor activity of antioxidant enzymes, and (iii) the presence of at least one female infertility factor does not alter TBARS levels or antioxidant activity. OBJECTIVE: Oxidative stress is an important cause of female infertility. During follicular development, oxidative stress can induce oocyte degeneration and apoptosis, modifying the meiotic spindle and causing lipid peroxidation, loss of membrane selectivity, inactivation of cytoplasmic enzymes, and DNA damage. The aim of this study was to verify relations between concentrations of lipid peroxides (TBARS) and activity of the antioxidant enzymes glutathione peroxidase (GPx) and catalase (CAT) with the occurrence of pregnancy, hormonal stimulation protocols and presence of a female infertility factor. DESIGN: Prospective study. MATERIALS AND METHODS: Follicular fluid from 146 patients submitted to ICSI was included. Groups were divided according to pregnancy, hormonal stimulation protocol (with or without LH), and presence or not of at least one female infertility factor. Follicular fluids were collected after oocyte retrieval, and TBARS levels and CAT and GPx activity analyzed by spectrophotometry. Student's T-test or ANOVA were used, and logistic models were calculated using embryo quality on day 3, presence of female infertility, and TBARS levels and CAT and GPx activity as independent variables, occurrence of pregnancy as a binary dependent variable. RESULTS: In the non-pregnant group an increase in GPx activity (p=0,04) was observed. In the other groups no differences were observed. Logistic regression produced a model which best predicted pregnancy including presence of female infertility factor, embryo quality on day 3 and GPx activity. Odds-ratio for GPx activity was 0.990 (increasing values of GPx activity decrease the odds of occurrence of pregnancy). CONCLUSIONS: We may conclude that (i) higher GPx activity is associated to negative outcomes in ICSI cycles, (ii) inclusion of LH in ovarian stimulation protocols does not alter TBARS concentrations nor activity of antioxidant enzymes, and (iii) the presence of at least one female infertility factor does not alter TBARS levels or antioxidant activity.

Could oxidative stress be responsible for poor oocyte competence in women with polycystic ovary syndrome?

OBJECTIVE: Circulating reactive oxygen species (ROS) have been reported to induce insulin resistance and hyperandrogenism in women with polycystic ovary syndrome (PCOS). Animal experiments have shown that oxidative stress (OS), induced in vitro, disturbs spindle organization during oocyte meiosis. This study aims to evaluate the effects of follicular fluid (FF) OS on meiotic spindle (MS) formation in oocytes and outcome of in vitro fertilization in PCOS women.DESIGN: Prospective randomized-controlled study at Institute of Reproductive Medicine.MATERIALS AND METHODS: 301 oocytes retrieved from 28 PCOS (Group-A) women and 218 oocytes from 32 age-matched non-PCOS (Group-B) women were analyzed. Long-protocol ovarian stimulation was undertaken and urinary human chorionic gonadotrophin (10,000IU) was used for final oocyte maturation. FF was collected from all follicles above 14mm. 217 FF samples of both groups containing mature oocytes were analyzed for ROS by chemiluminescence (luminometer). All mature oocytes were denuded and observed under Polscope to visualize MS. FF samples of Gr-A were classified as Gr-A1 (oocyte with MS) and Gr-A2 (oocyte without MS). Intracytoplasmic Sperm Injection was performed in all oocytes. After 16-17 hours of injection, pronuclei were scored by revised Z-scoring system. Embryo grading, 48 hours following oocyte retrieval, was done by Veeck's classification. A maximum of 3 embryos were transferred on D2. Gestational sac demonstrated on transvaginal sonography was considered as clinical pregnancy. Statistical comparisons were performed by Student's t-test and Chi Square Test. P<0.05 was considered as statistically significant.RESULTS: The average number of oocytes recovered were significantly (p<0.001) higher in Gr-A (12.08±2.15) compared to Gr-B (6.81±2.16). Oocytes without MS were significantly higher (p<0.001) in Gr-A (3.5±1) than Gr-B (1.62±1.14). In Gr-A, ROS (101.70±19.09) were observed to be significantly (P≤0.001) higher than Gr-B (88.91±25.34). Interestingly, ROS levels were also significantly (p<0.001) less in Gr-A1 (62.37±20.18) compared to Gr-A2 (120.14±17.28). An average of 3.5±0.9 and 3.06±1.06 zygotes (Z1/Z2) were observed in PCO and normal women respectively. However, the number of high-grade embryos was not significantly different (2.9±1.08 and 2.56±1.03). The clinical pregnancy rates were significantly higher (p=0.02) in Gr-B (37.5%) than Gr-A (8.3%).CONCLUSIONS: Higher oxidative stress in FF of PCOS women appears to inhibit MS formation and hence affect embryo quality and pregnancy rates. OBJECTIVE: Circulating reactive oxygen species (ROS) have been reported to induce insulin resistance and hyperandrogenism in women with polycystic ovary syndrome (PCOS). Animal experiments have shown that oxidative stress (OS), induced in vitro, disturbs spindle organization during oocyte meiosis. This study aims to evaluate the effects of follicular fluid (FF) OS on meiotic spindle (MS) formation in oocytes and outcome of in vitro fertilization in PCOS women. DESIGN: Prospective randomized-controlled study at Institute of Reproductive Medicine. MATERIALS AND METHODS: 301 oocytes retrieved from 28 PCOS (Group-A) women and 218 oocytes from 32 age-matched non-PCOS (Group-B) women were analyzed. Long-protocol ovarian stimulation was undertaken and urinary human chorionic gonadotrophin (10,000IU) was used for final oocyte maturation. FF was collected from all follicles above 14mm. 217 FF samples of both groups containing mature oocytes were analyzed for ROS by chemiluminescence (luminometer). All mature oocytes were denuded and observed under Polscope to visualize MS. FF samples of Gr-A were classified as Gr-A1 (oocyte with MS) and Gr-A2 (oocyte without MS). Intracytoplasmic Sperm Injection was performed in all oocytes. After 16-17 hours of injection, pronuclei were scored by revised Z-scoring system. Embryo grading, 48 hours following oocyte retrieval, was done by Veeck's classification. A maximum of 3 embryos were transferred on D2. Gestational sac demonstrated on transvaginal sonography was considered as clinical pregnancy. Statistical comparisons were performed by Student's t-test and Chi Square Test. P<0.05 was considered as statistically significant. RESULTS: The average number of oocytes recovered were significantly (p<0.001) higher in Gr-A (12.08±2.15) compared to Gr-B (6.81±2.16). Oocytes without MS were significantly higher (p<0.001) in Gr-A (3.5±1) than Gr-B (1.62±1.14). In Gr-A, ROS (101.70±19.09) were observed to be significantly (P≤0.001) higher than Gr-B (88.91±25.34). Interestingly, ROS levels were also significantly (p<0.001) less in Gr-A1 (62.37±20.18) compared to Gr-A2 (120.14±17.28). An average of 3.5±0.9 and 3.06±1.06 zygotes (Z1/Z2) were observed in PCO and normal women respectively. However, the number of high-grade embryos was not significantly different (2.9±1.08 and 2.56±1.03). The clinical pregnancy rates were significantly higher (p=0.02) in Gr-B (37.5%) than Gr-A (8.3%). CONCLUSIONS: Higher oxidative stress in FF of PCOS women appears to inhibit MS formation and hence affect embryo quality and pregnancy rates.

Effect of Oxidative Stress in Follicular Fluid and Serum on the Outcome of Assisted Reproductive Procedures

1) To measure levels of lipid peroxidation (LPO) and superoxide dismutase (SOD) and catalase (CAT) in the serum and follicular fluid from patients undergoing IVF; 2) to examine the association among LPO, SOD and CAT with oocyte maturity, embryo quality, fertilization, cleavage, and pregnancy rates. Prospective study at a tertiary care medical center The Institutional Review Board approved this study. Seventy-three patients undergoing IVF participated in this study. Clear follicular fluid and blood serum samples were collected at oocyte retrieval. Follicular fluid LPO levels were measured by the thiobarbituric acid method and SOD and CAT were determined with a spectrophotometer. The activity of the SOD was based on the adrenocromo concentration, resulting from the adrenaline oxidation by the radical superoxide. Catalase activity was determined by the velocity of hydrogen peroxide consumed. The mean and the standard error for SOD (4.45 ± 1.4) and CAT (1.7 ± 0.28) in the follicular fluid were higher than the levels in the blood serum (3.6 ± 0.5; p = 0.04, and 1.1 ± 0.13; p = 0.03, respectively). The mean and standard error for the LPO levels were lower in the follicular fluid (2.4 ± 0.15) than the blood serum (4.2 ± 0.4; P = 0.04). The SOD enzymatic activity had a significant correlation with the oocyte fertilization rates (r = 3.69; p = 0.013). There were no correlations between the LPO, SOD and CAT with the cleavage and embryo quality. Pregnant patients were significantly younger than the patients who did not become pregnant (33.5 ± 4.2 years vs. 37.9 ± 5.2 years). After adjusting for age, a statistical significant correlation was detected between the LPO levels in the blood serum (r = 7.3; p = 0.01) and follicular fluid (r = 3.6; p = 0.05) with the pregnancy rates. There were no correlations between the SOD and CAT with the pregnancy outcome. SOD levels had a significant correlation with the fertilization rate and the levels of levels were positively correlated with the pregnancy rate. Markers of oxidative stress may represent metabolic activity within the follicle, and some amounts may be necessary to establish a pregnancy.

Effect of oxidative stress in follicular fluid on the outcome of assisted reproductive procedures

Objective(s) To measure levels of lipid peroxidation (LPO) and total antioxidant capacity (TAC) in the follicular fluid from patients undergoing IVF and to examine the association among LPO and TAC and oocyte maturity, embryo quality, fertilization, cleavage, and pregnancy rates. Design Prospective study. Setting Tertiary care medical center. Patient(s) Forty-one patients undergoing IVF. Intervention(s) Clear follicular fluid specimens were collected at oocyte retrieval. Main outcome measure(s) Follicular fluid LPO and TAC levels measured by the thiobarbituric acid and the chemiluminescence method, respectively. Result(s) The mean LPO level was 0.95 μmol malonaldehyde/L, and the mean TAC level was 819.16 Trolox equivalents. No correlation was seen among LPO or TAC levels and oocyte maturity, fertilization, cleavage, and embryo quality. The pregnancy rate was 31.7%. Pregnant patients were significantly younger than the patients who did not become pregnant (30.53 ± 3.5 years vs. 34.32 ± 3.98 years). After adjusting for age, a positive correlation was seen between pregnancy rate and LPO and between pregnancy rate and TAC. Conclusion(s) Both LPO and TAC levels were positively correlated with the pregnancy rate. Lipid peroxidation may be a good marker of metabolic activity within the follicle, and some amounts may be necessary to establish a pregnancy.