Oxidative Stress In Follicular Fluid Research Articles

Obesity and follicular fluid oxidative stress: Relationship to ICSI outcome

Abstract Objective To study the influence of female obesity on follicular fluid oxidative stress and to correlate it to intracytoplasmic sperm injection (ICSI) outcome. Study design Seventy-four normal females below the age of 40 undergoing ICSI for reason of male factor infertility were enrolled in the study. They were divided into 2 groups according to body mass index (BMI); Group I (non-obese) ( n = 24, BMI 2 ) and Group II (obese) ( n = 50, BMI ⩾ 25 Kg/m 2 ). Oxidative stress markers (MDA (malonaldehyde), NO 2 /NO 3 (Nitrite/Nitrate) ratio, GSH (reduced glutathione) and GSH/GSSG (reduced glutathione/oxidized glutathione) ratio) were measured by high performance liquid chromatography (HPLC). Results Obese women had significantly higher mean follicular fluid MDA ( P = 0.006) as well as NO 2 /NO 3 ratio ( P = 0.004). BMI strongly correlated to follicular fluid MDA ( P = 2 /NO 3 ratio ( P = 0.02). GSH and GSH/GSSG ratio showed a non-significant difference between the two groups ( P = 0.14 and 0.67, respectively). Clinical pregnancy rate was significantly higher in the non-obese group (87%) compared to the obese (43%) ( P = P = Conclusion Obesity aggravates follicular fluid oxidative stress with a negative impact on pregnancy outcome of ICSI cycles.

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.

Correlations of follicular fluid oxidative stress biomarkers and enzyme activities with embryo morphology parameters during in vitro fertilization

To measure antioxidant enzyme activities and lipid peroxidation levels within follicular fluid (FF) and evaluate correlations with early embryo quality. Individual FF samples were obtained prospectively on the day of oocyte collection and assessed for lipid peroxidation as specific positional isomers of hydroperoxy and hydroxy fatty acids by high-performance liquid chromatography and antioxidant enzyme activities by automated kinetic enzyme assays. Spearman rank correlation coefficients, adjusted for age and day of transfer, were used to assess associations between antioxidant enzymes and lipid peroxidation products and embryo quality using a 1 follicle-1 oocyte/embryo approach. Post hoc power analysis was conducted to help interpret null results. A university clinic. Thirty-nine women undergoing IVF. None. Embryo cell number and embryo fragmentation score (EFS) at transfer. No significant correlations between lipid peroxidation derivatives or antioxidant enzyme activities and embryo quality were obtained. Post hoc power analysis indicated possible undetected associations between EFS and 13-hydroxy octadecatrienoic acid and 13-hydroperoxy octadecadieneoic acid. Our preliminary dataset suggests that most lipid peroxidation products and antioxidant enzyme activities within FF are not associated with the quality of embryos, using EFS and embryo cell number as end points. However, further consideration of associations between EFS and 13-hydroxy octadecatrienoic acid and 13-hydroperoxy octadecadieneoic acid is warranted.

Oxidative stress markers in follicular fluid (FF) and assisted reproduction outcomes in patients with infertility due to polycystic ovary syndrome (PCOS) or to tubal and/or male factor

OBJECTIVE: Oxidative stress (OS) is related with worse oocyte quality in PCOS patients, but little is known about follicular environment and assisted reproduction outcomes. The objective was to compare OS markers in the follicular fluid (FF) of non-obese infertile PCOS women and non-obese ovulatory patients with infertility due to tubal and/or male factor (control) submitted to ovarian stimulation (COS) for intracytoplasmic sperm injection (ICSI). We also compared ovarian stimulation response and ICSI results. DESIGN: Prospective study. MATERIALS AND METHODS: The study was conducted on 16 cycles of 15 PCOS patients and 66 cycles of 57 infertile patients due to tubal and/or male factor submitted to COS for ICSI. FF was aspirated from 13 PCOS patients and 40 controls. Malondialdehyde, hydroperoxides, advanced oxidation protein products, glutathione and vitamin E were determined by absorbance and high performance liquid chromatography. The ovarian stimulation response and ICSI results were evaluated in both groups. RESULTS: No significant differences in OS markers levels in FF and ICSI outcomes were observed between the groups (Table 1).Table 1FF levels of OS markers and ICSI outcomes from infertile PCOS and controls.ResultsPCOSControlPMDA (μmol/g pt)23,0 ± 6,321,0 ± 4,40,09AOPP (μmol/L)112,1 ± 59,7107,3 ± 38,90,84FOX (μmol/g pt)5,4 ± 0,45,8 ± 0,70,15GSH (nmol/L)279,1 ± 59,7265,5 ± 71,00,27Vitamin E (μmol/L)8,5 ± 4,68,6 ± 2,80,71Number of produced embryos2,6 ± 1,43,1 ± 1,10,23Clinical Pregnancy rate (%)31,2 (5/16)24,2 (16/66)0,54MDA:Malondialdehyde, FOX:hydroperoxides, AOPP:advanced oxidation protein products, GSH:glutathione. Open table in a new tab MDA:Malondialdehyde, FOX:hydroperoxides, AOPP:advanced oxidation protein products, GSH:glutathione. CONCLUSION: The present study didn't demonstrate OS in FF or worse ICSI outcomes in non-obese, oligomenorrheic, and infertile PCOS women compared to infertile ovulatory controls submitted to controlled ovarian stimulation for ICSI.

Effect of Follicular Fluid Oxidative Stress on Meiotic Spindle Formation in Infertile Women with Polycystic Ovarian Syndrome

Background: The effect of follicular fluid (FF) oxidative stress (OS) on meiotic spindle (MS) formation in oocytes and subsequent outcome in women with polycystic ovarian syndrome (PCOS) are evaluated in this study. Methods: 326 oocytes from 35 PCOS women (group A) and 208 oocytes from 32 women with tubal infertility (group B) were visualized for MS using PolScope. FF was analyzed for OS markers including reactive oxygen species (ROS), lipid peroxidation and total antioxidant capacity (TAC). Group A was further classified into groups A1 and A2, and group B into groups B1 and B2 depending upon the presence or absence of MS, respectively. Results: MS formation was absent in a significantly higher number of oocytes in group A compared to group B (p ≤ 0.05). OS markers were significantly higher in group A compared to group B (p ≤ 0.05). Fertilization rate, number of good quality embryos and clinical pregnancy rates were higher in group B compared to group A, though not statistically significant. FF ROS was significantly higher and TAC significantly lower in groups A2 and B2 compared to groups A1 and B1 (p < 0.001). Conclusion: The presence of MS and oocyte maturation in PCOS women showed a positive correlation with low levels of OS.

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.

Evaluation of the relationship between follicular fluid oxidative stress, ovarian hormones, and response to gonadotropin stimulation

To investigate the relationship between oxidative stress and the underlying causes of infertility, preovulatory ovarian hormones, and ovarian response to gonadotropin stimulation in patients undergoing assisted reproductive techniques. Prospective, cross-sectional study. Assisted conception unit, university hospital. One hundred thirty women presenting with infertility, of the following types: male factor (n = 56), unexplained (n = 36), tubal factor (n = 16), polycystic ovary syndrome (n = 15), and endometriosis (n = 7). Follicular fluid (FF) and peripheral blood samples were collected at oocyte retrieval. Blood and FF samples were analyzed for inhibin A, inhibin B, activin A, anti-Müllerian hormone, and E(2) by using ELISA. Total antioxidant capacity (TAC) was measured in plasma and FF by using a calorimetric microplate assay. There was no significant relationship between plasma or FF TAC and the underlying etiology of infertility. There was a statistically significant positive association between FF E(2) levels and TAC (r = 0.26). Higher antral follicle count, delta E(2) (day 3 E(2) minus day 2 E(2)), preovulatory serum anti-Müllerian hormone, inhibin B, and E(2) were associated with good ovarian response, whereas higher FF E(2) was associated with a statistically significant poor response. No significant direct relationship was observed between TAC and ovarian response as well as between TAC or any of the parameters measured and pregnancy outcome. Oxidative stress has an impact on the production of granulosa cell steroid hormones, in particular E(2), which is an important predictor of ovarian response. The positive association between FF E(2) and total antioxidant capacity suggests that E(2) may play a role in the ovarian antioxidant-oxidant balance.

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.

Relationship of the Follicular Fluid Oxidative Stress Parameters and the Outcome of Intracytoplasmic Sperm Injection

The purpose of this study was to assess the relationship between the levels of reactive oxygen species (ROS), total antioxidant capacity (TAC) and ROS-TAC score in the follicular fluid of women undergoing IVF/ICSI and the success of pregnancy in the respective cycles.

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.