Follicular Fluid Vascular Endothelial Growth Factor Research Articles

VEGF Concentration in a Preovulatory Leading Follicle Relates to Ovarian Reserve and Oocyte Maturation during Ovarian Stimulation with GnRH Antagonist Protocol in In Vitro Fertilization Cycle.

Serum vascular endothelial growth factor (VEGF) is involved in follicular vascularization, oxygenation, and consequently in oocyte maturation and embryo development. Unanswered questions remain regarding the relationship of intrafollicular VEGF level in preovulatory leading follicles to oocyte maturation and ovarian reserve during ovarian stimulation. We conducted this study to investigate the relationship of intrafollicular VEGF level in the fluid of single preovulatory leading follicles to ovarian reserve and oocyte maturation in patients receiving GnRH antagonist in vitro fertilization (IVF) protocol treatment. One hundred and eighty-five patients receiving IVF treatment were recruited and assigned to low-, normal-, and high-ovarian-reserve groups according to their serum anti-Müllerian hormone (AMH) level. Follicular fluid (FF) in preovulatory leading follicles, serum profiles, and clinical variables were collected for analysis. The result disclosed a significant among-group difference in FF VEGF concentration. Moreover, the serum AMH level was also negatively correlated with FF VEGF level. The oocyte maturation rate tended to be increased at higher AMH levels. FF VEGF concentration was significantly positively correlated with basal FSH level. In conclusion, FF VEGF concentration has a negative association with ovarian reserve level and oocyte maturation rate in patients undergoing GnRH antagonist IVF protocols.

Plasma and follicular fluid osteopontin levels during ovarian cycle and their correlation with follicular fluid vascular endothelial growth factor levels

Osteopontin (OPN) is a multifunctional secreted glycoprotein. We evaluated OPN concentrations in blood and follicular fluid (FF) during the ovarian cycle and their relationship with the production of vascular endothelial growth factor (VEGF), which is involved in the pathophysiology of ovarian hyperstimulation syndrome (OHSS). Twenty-two women undergoing in vitro fertilization (minimal stimulation protocol with clomiphene citrate) were enrolled. Samples were collected (a) on the third day of withdrawal bleeding, (b) 2 days before oocyte retrieval, and (c) on the day of oocyte retrieval. FF was collected during oocyte retrieval. The OPN concentration in each specimen and the VEGF concentration in FF was measured by enzyme-linked immunosorbent assays. Plasma OPN concentrations were (in ng/mL): (a) 416 ± 37.2, (b) 378 ± 35.8, and (c) 390 ± 40.0, with no significant differences between the groups. The OPN concentration in FF was 106 ± 13.4 ng/mL. A positive correlation was found between OPN concentrations in FF and plasma samples. A positive correlation was also found between plasma OPN and FF VEGF concentrations, irrespective of the blood-sampling period. Plasma OPN concentration is suggested to reflect the FF VEGF level at oocyte retrieval and maybe a novel clinical marker for predicting the risk for OHSS.

Investigating apoptotic, inflammatory, and growth markers in poor responders undergoing natural in vitro fertilization cycles: a pilot study.

This study investigates follicular fluid (FF) from patients with poor and normal ovarian response undergoing natural assisted reproductive technology cycles. We report about (1) cell-free DNA (cfDNA), which reflects apoptosis; (2) corticotropin-releasing hormone (CRH); (3) interleukin (IL)-15, which reflects inflammation; (4) granulocyte colony-stimulating factor (G-CSF); (5) vascular endothelial growth factor (VEGF); and (6) insulin-like growth factor I (IGF-I), which reflects follicular growth. Forty-four poor responders and 44 normal responders-according to the Bologna criteria-were recruited. FF samples were prepared for cfDNA quantification employing Q-PCR and for CRH, IL-15, G-CSF, VEGF, and IGF-I quantification employing ELISA. Statistically nonsignificant different levels of FF cfDNA, CRH, IL-15, VEGF, and IGF-I were observed. Interestingly, statistically significant higher G-CSF levels were observed in normal responders (302.48 ± 474.36 versus 200.10 ± 426.79 pg/mL, P=0.003). Lower cfDNA integrity was observed in cycles resulting in clinical pregnancy for both groups (normal: 0.07 ± 0.04 versus 0.25 ± 0.17 ng/μL, P < 0.001; poor: 0.10 ± 0.06 versus 0.26 ± 0.12 ng/μL, P < 0.001). The results predominantly showcase similarities between normal and poor responders pertaining to inflammatory, apoptotic, and growth factors. This may be attributed to the employment of natural cycles in order to exclude controlled ovarian stimulation as a factor-indicating its detrimental effect. As G-CSF levels presented significantly higher in normal responders, its vital role in understanding a compromised ovarian response is highlighted.

The expression of angiotensin II receptors mRNA in granulosa-lutein cells in endometriosis patients who underwent ovarian surgery before in vitro fertilization

Angiogenesis is important to folliculogenesis and oocyte maturation. Vascular endothelial growth factor (VEGF) in follicular fluid (FF) induces folliculogenesis. Angiotensin II is involved in angiogenesis through angiotensin II receptors. To elucidate the mechanisms of oocyte maturation disorders in endometriotic patients, the authors investigated the concentration of VEGF, angiotensin II, and soluble receptor of VEGF (sFlt-1) in FF and the expression of angiotensin II receptors mRNA in granulosa-lutein (GL) cells in endometriosis patients with controlled ovarian stimulation. FF samples were obtained from 56 IVF cycles (control = 47, endometriosis = 9). GL cells were obtained from 15 IVF cycles (control = 10, endometriosis = 5). The authors examined the concentration of VEGF, sFlt-1, and angiotensin II in FF by ELISA, and the expression of angiotensin II receptors mRNA by quantitative RT-PCR. Informed consents were obtained from all patients and IRB approval was obtained. The expression levels of angiotensin II type 1 and type 2 receptor mRNA in endometriosis was significantly lower than the control. The number of immature oocytes was significantly higher in endometriosis than in control. Lower expression of angiotensin II receptors may affect folliculogenesis and oocyte maturation.

Levels of EG-VEGF and VEGF in serum and in the follicular fluid on the day of oocyte retrieval and reproductive outcome among IVF patients

The aim of this retrospective, observational, case control study was to investigate the correlation of endocrine gland-derived vascular endothelial growth factor (EG-VEGF) and vascular endothelial growth factor (VEGF) concentrations in serum and follicular fluid (FF) in women undergoing controlled ovarian hyperstimulation (COH) protocols and IVF/ICSI with the reproductive outcome. When the authors compared levels of EG-VEGF and VEGF in FF and serum, they did not observe differences in the group without pregnancy compared to the group of patients with clinical pregnancy (EG-VEGF, respectively, p = 0.12, p = 0.94, and p = 0.16 and VEGF p = 0.16, p = 0.6, and p = 0.32). The authors found lower levels of EG-VEGF in FF from large follicles in the group of patients with clinical pregnancy who miscarried compared to the group of patients who had no pregnancy (p = 0.04); when they compared this group with the group of patients who delivered a baby, there was no statistical significance. Lower levels of EG-VEGF in the follicular fluid from large follicles are negatively correlated with positive outcome of IVF, and correlate with miscarriage rates in patients after IVF.

Follicular fluid Vascular Endothelial Growth Factor (VEGF) could be a predictor for pregnancy outcome in normo-responders and polycystic ovary syndrome women undergoing IVF/ICSI treatment cycles

Abstract Objective: The aim of this study was to evaluate the serum and follicular fluid levels of Vascular Endothelial Growth Factor (VEGF) as possible predictive markers for IVF outcome in PCOS patients. Design: Prospective cross-sectional study. Materials and Methods: Forty patients with PCOS and 40 age and weight matched control women were included in this study. All of them were enrolled in the IVF program. Blood and follicular fluid (FF) samples were collected from all subjects on the day of oocyte retrieval. VEGF levels were measured in serum and follicular fluid (FF) using enzyme linked immunosorbent assay (ELISA). The correlation between VEGF levels and number of retrieved oocyte, number of mature oocyte, number of fertilized oocyte, number of embryos, percent of mature oocytes, fertilization rate and pregnancy was assessed. Results: PCOS patients showed higher serum VEGF levels than controls (297.46 ± 159.45 pg/ml vs. 134.08 ± 113.06 pg/ml, p value = 0.0001) and higher FF VEGF levels compared to controls (1358.92 ± 820.86 pg/ml vs. 987.58 ± 236.62 pg/ml, p value = 0.007). Serum VEGF levels were not significantly correlated with the number of retrieved oocyte in both PCOS ( r = 0.37, p value = 0.19) and control women ( r = −0.03, p value = 0.83), and FF VEGF levels were not significantly correlated with the number of retrieved oocyte in both PCOS ( r = −0.06, p value = 0.7) and control women ( r = −0.13, p value = 0.43). Both serum and FF VEGF levels were not significantly correlated with the number of mature oocyte, number of fertilized oocyte, number of embryos, percent of mature oocytes or fertilization rate. Serum VEGF levels were not significantly different between pregnant and non-pregnant women in both PCOS (256.69 ± 139.11 pg/ml vs. 300.20 ± 114.60 pg/ml, p value = 0.416) and control women (107.98 ± 115.51 pg/ml vs. 154.39 ± 111.09 pg/ml, p value = 0.271), whereas FF VEGF levels were significantly lower in FFs from the group of pregnancy than those from the group of no-pregnancy in both PCOS (812.04 ± 308.16 pg/ml vs. 1936.78 ± 771.79 pg/ml, p value = 0.0001) and control women (822.45 ± 199.45 pg/ml vs. 1171.98 ± 139.60 pg/ml, p value = 0.0001). FF VEGF levels had a significant predictive value of pregnancy in both PCOS ( p value = 0.0001) and control women ( p value = 0.0001). Conclusion: FF VEGF levels may serve as a reliable predictive marker for pregnancy in both normal women and women with polycystic ovary syndrome undergoing IVF.

Assessment of follicular and serum VEGF and IGF-1 in ICSI patients: hMG vs rFSH

Purpose: To investigate the effect of recombinant follicular stimulating hormone (r-FSH) and human menopausal gonadotropin (hMG) on follicular microenvironment via assessment of follicular and serum vascular endothelial growth factor (VEGF) and insulin-like growth factor-1 (IGF-1) levels in intracytoplasmic sperm injection (ICSI) cycles. Materials and Methods: Designed as a prospective cohort study. Twenty-five patients underwent controlled ovarian hyperstimulation (COH) with r-FSH and 20 patients underwent with hMG. Results: Both groups were comparable regarding the women’s mean age and body mass index (BMI). The amount of VEGF (pg/ml) in serum and follicular fluid in the group I and II were comparable (275 &#x00B1; 135.3 vs 330.7 &#x00B1; 190.0; p > 0.05 and 2,081.1 &#x00B1; 1095.1 vs 1,971.1 &#x00B1; 975.6; p > 0.05, respectively). The amount of IGF-1 (ng/ml) in serum and follicular fluid in the group I and II were also comparable (225.3 &#x00B1; 69.3 vs 204.1 &#x00B1; 56.3, p > 0.05 and 176.1 &#x00B1; 67.2 vs 185.8 &#x00B1; 48.7, p > 0.05, respectively). Pregnancy rates were also comparable between groups. Conclusions: The hMG and r-FSH in COH produced comparable follicular microenvironment regarding follicular VEGF and IGF-1.

Follicular fluid and serum vascular endothelial growth factor, interleukin (IL)-1β and glycodelin concentrations: comparison between long-gonadotropin-releasing hormone (GnRH)-agonist and GnRH-antagonist cycles: a randomized controlled trial

In this randomized controlled trial, we aimed to examine whether differences exist among patients who underwent assisted reproductive technology treatment with a long-GnRH-agonist compared to a GnRH-antagonist protocol in terms of levels of follicular fluid (FF) and serum concentrations of vascular endothelial growth factor (VEGF), glycodelin and interleukin (IL)-1β on the day of oocyte pick-up (OPU). In 80 infertile couple with male factor or unexplained infertility, 40 women stimulated with GnRH-antagonist protocol and 40 women with the long-GnRH-agonist protocol. FF and blood serum samples were obtained simultaneously from 80 women during the OPU procedure and the concentrations of VEGF, IL-1β and glycodelin were measured with commercially available kits. Concentrations of FF VEGF, IL-1β and glycodelin were not significantly different in the long-GnRH-agonist and GnRH-antagonist groups, and neither were serum concentrations of VEGF, IL-1β and glycodelin. According to our results in at least, we can say that minor differences between these protocols in terms of clinical pregnancy do not depend on VEGF, glycodelin or IL-1β.

Effects of EG-VEGF, VEGF and TGF-β1 on pregnancy outcome in patients undergoing IVF-ET treatment

To investigate the correlation of endocrine gland-derived vascular endothelial growth factor (EG-VEGF), vascular endothelial growth factor (VEGF) and transforming growth factor beta 1 (TGF-β1) with the corresponding reproductive outcome in patients who received in vitro fertilization-embryo transfer (IVF-ET). Sixty-seven women undergoing IVF-ET at a university tertiary hospital were recruited for a prospective study. Concentrations of EG-VEGF, VEGF and TGF-β1 were measured by enzyme-linked immunosorbent assay (ELISA) in follicular fluid (FF) collected during oocyte retrieval (OR) and in serum collected 2 days after OR. In FF, concentrations of both EG-VEGF and VEGF were negatively correlated with peak E2 and the number of MII oocytes retrieved, and positively correlated with each other. In serum, concentrations of all the three growth factors were positively correlated with the rate of good quality embryo, and with one another. Patients in the pregnancy group had lower peak E2 concentrations and higher serum EG-VEGF concentrations than those in the non-pregnancy group, but such tendency was not observed in the case of VEGF and TGF-β1. Both concentrations of EG-VEGF and VEGF in FF were negatively correlated with ovarian response and oocyte maturation. Concentrations of all the three growth factors in serum were positively correlated with embryo quality, but only serum concentrations of EG-VEGF were associated with the pregnancy outcome.

Agonist and antagonist coast

The use of GnRH-a in ovarian stimulation permitted stronger stimulation resulting in an increased incidence of OHSS. The first Cochrane review comparing GnRH agonist and GnRH antagonist protocols for ovarian stimulation showed no significant difference in OHSS rate between the two protocols, however, a recent Cochrane review showed a highly significant decrease in the incidence in OHSS rate in the antagonist protocol. Coasting is a commonly used procedure for preventions of OHSS. The optimum time to start coasting is when the lead follicle reaches 16 mm in diameter and hCG should be given when E2 level drops below 3000 pg/ml. Coasting may act by diminishing the functioning granulosa cell cohort. Administration of daily GnRH antagonist in high risk patients for OHSS who were down-regulated by GnRH-a resulted in rapid drop of E2 and decrease in incidence of OHSS. A series of patients who developed early OHSS were treated by daily GnRH antagonist injections, all embryos were cryopreserved. No progression to severe OHSS was observed.

In vitro viability and secretory capacity of human luteinized granulosa cells after gonadotropin-releasing hormone agonist trigger of oocyte maturation

To evaluate viability of luteinized granulosa cells obtained from patients triggered with either GnRH agonist or hCG and to assess the secretion of steroids and vascular endothelial growth factor (VEGF) by cultured luteinized granulosa cells in the presence or absence of hCG. Prospective, randomized controlled trial. University-based fertility center. A subset of patients who underwent a randomized trial involving GnRH agonist trigger after GnRH antagonist protocol vs. hCG trigger after pituitary suppression with GnRH agonist protocol. In vitro fertilization cycles. Proportion of apoptosis; basal and hCG-induced secretion of E(2), P, and VEGF by luteinized granulosa cells; follicular-fluid VEGF and luteal-phase serum E(2), P, and plasma VEGF concentrations. There were no differences in the proportion of granulosa/luteal cell apoptosis, follicular-fluid or luteal-phase plasma VEGF concentration, or basal culture media E(2), P, and VEGF concentrations between the two groups. Addition of hCG to the culture media significantly increased the P concentration in both groups, but there were no changes in E(2) or VEGF concentrations. Serum E(2) levels were lower at 5 and 9 days after GnRH agonist compared with hCG trigger. The granulosa/luteal cells obtained on the day of oocyte retrieval after GnRH agonist trigger are still viable and have the capacity to respond to hCG by increasing the secretion of steroids.

Decreased concentrations of pigment epithelium–derived factor in peritoneal fluid of patients with endometriosis

To determine whether patients with endometriosis have altered levels of pigment epithelium-derived factor (PEDF) in peritoneal fluid, concentrations of PEDF in peritoneal fluid collected from 42 patients with endometriosis and 30 patients without endometriosis were measured with enzyme-linked immunosorbent assay. We detected significantly lower levels of peritoneal fluid PEDF in patients with endometriosis compared with patients without endometriosis, suggesting that peritoneal fluid PEDF plays a role in the pathogenesis of this disorder.

Increased follicular fluid inhibin A in infertile patients with minimal/mild endometriosis submitted to in vitro fertilization (IVF) using a minimal stimulation protocol

OBJECTIVE: This study aims to determine the follicular fluid concentration and the role of vascular endothelial growth factor (VEGF) and inhibin A in infertile patients with minimal/mild endometriosis (EDT) undergoing IVF using a minimal stimulation protocol. DESIGN: We did a cross-sectional study. MATERIALS AND METHODS: We included 81 infertile patients submitted to IVF. Control group was composed of 47 infertile patients without endometriosis and the endometriosis group encompassed 37 infertile women with minimal/mild EDT. IVF treatment was done using a minimal stimulation protocol (gonadotrophin and GnRH antagonist). We measured VEGF and Inhibin A after oocyte pickup from all cycles with specific commercial kits. We considered statistic significance when P < 0.05, using t test or qui-square. RESULTS: The groups were comparable in terms of age, body mass index and infertility characteristics. Follicular fluid VEGF was similar between the groups (1123±121 pg/ml versus 1194±134 pg/ml, P=0.699). However, patients with EDT presented a higher follicular level of inhibin A (2595±191 pg/ml) as compared to those without EDT (1915±212 pg/ml, P=0.02). In order to certify that our results were not obtained only by chance we performed a regression analysis considering follicular content of inhibin A as a dependent variable and controlling for: 1) minimal stimulation protocol and 2) presence of EDT. We confirmed that only the presence of EDT was associated with an increased follicular fluid concentration of inhibin A (P=0.026). CONCLUSIONS: We clearly showed for the first time, using a robust statistical analysis, that endometriosis was associated with an increased follicular fluid concentration of inhibin A. In addition, we refute the hypothesis that this disease was associated with any change of VEGF in the follicular fluid. The exact mechanism was not well determined, but could be related to several ovarian-dysfunction presented by those infertile patients with endometriosis.

The measurements of RAGE, VEGF, and AGEs in the plasma and follicular fluid of reproductive women: the influence of aging

To examine the advanced glycation end products (AGEs), the soluble isoform of the receptor for AGEs (sRAGE), and vascular endothelial growth factor (VEGF) concentrations in plasma and follicular fluid (FF) from reproductive-age women. Clinical preliminary study based on the regulations of the ethical committee at National Center for Child Health and Development (NCCHD). Women's health clinical office at NCCHD and Aska clinic. Reproductive-age women, young group (<35 yrs) and old group (>or=35 yrs), who agreed to let us use plasma or FF samples for the measurements of AGEs, sRAGE, and VEGF. Measurements of AGEs, sRAGE, and VEGF in plasma and FF by ELISA to examine the difference by aging and reproductive dysfunction. The plasma concentration of sRAGE was significantly higher in the young group; VEGF in FF was significantly higher in the old group. sRAGE in FF showed a tendency of positive correlation with the number of oocytes. The plasma sRAGE concentration was significantly correlated positively with FF sRAGE and inversely with FF VEGF at the time of egg collection. The measurement results suggest a possibility that RAGE-VEGF regulation may be related to reproductive dysfunction in aging women, and that plasma sRAGE might be a biologic marker of reproductive condition.

Vascular endothelial growth factor and its soluble receptor in patients with polycystic ovary syndrome undergoing IVF

We aimed to examine the behaviour of the angiogenetic factor vascular endothelial growth factor (VEGF) and its soluble receptor (sVEGFR-1) in polycystic ovary patients undergoing In vitro fertilisation (IVF) compared with respect to normally ovulating controls. Levels of VEGF and sVEGFR-1 were compared in follicular fluid and serum, both on the day of human choriogonadotropin (hCG) administration and on the day of oocyte retrieval (OR), in controls and polycystic ovarian syndrome (PCOS) patients undergoing IVF cycles. The bioactivity of VEGF (VEGF/sVEGFR-1 ratio) in the two groups was calculated.Thirty PCOS patients and 20 controls referring to the IVF Centre of the University of Pisa (Italy) were enrolled. In each patient, blood samples were collected on the day of hCG and on the day of OR administration, and follicular fluid samples. VEGF and sVEGFR-1 were measured by Enzyme Linked Immuno Sorbant Assay (ELISA).Serum VEGF bioactivity markedly increased in both groups after hCG administration. Serum and follicular fluid VEGF bioactivity was greater in PCOS patients than in controls on the day of OR. The increase in VEGF bioactivity in PCOS patients undergoing IVF was not only because of increasing levels of VEGF but also to decreasing levels of its soluble receptor. We believe that additional studies will clarify their role in the pathogenesis of ovarian hyperstimulation syndrome, which most often occurs in patients with PCOS.

Vascularity and expression of angiogenic factors in bovine dominant follicles of the first follicular wave1

To determine the relationships among vascularity, expression of angiogenic factors, and selected intrafollicular factors in dominant and nondominant follicles of the first follicular wave, ovaries were obtained on d 3 of the estrous cycle from mature cross-bred beef heifers (n = 8) after a synchronized estrus. Follicular fluid (FF) was collected from all follicles > or = 3 mm for determination of estradiol-17beta (E), progesterone (P4), vascular endothelial growth factor (VEGF), and IGFBP concentrations. The ovaries were then perfusion-fixed and used for histochemical detection of lectin BS-1 (a marker of endothelial cells and thus vascularization) binding, and immunolocalization of VEGF, endothelial nitric oxide synthase (eNOS), and proliferating cell nuclear antigen, followed by image analysis of selected follicles. Follicles were classified, based on E and P4 concentrations in FF, as dominant, estrogen-active (EA; E:P4 > or = 1) or nondominant, estrogen-inactive (EI; E:P4 <1). Concentrations of E and VEGF in FF, the area of positive staining for lectin BS-1, VEGF, and eNOS, and the labeling index (an index of the percentage of cells proliferating) in granulosa and theca layers were greater (P < 0.05) in the EA than in the EI follicles, but concentrations of P4 and IGFBP in FF were less (P < 0.05) in EA than in EI follicles. In addition, vascularity was positively correlated (P < 0.05) with VEGF and eNOS protein expression, and tended (P < 0.1) to be positively correlated with the E:P4 ratio in FF but tended (P < 0.1) to be negatively correlated with IGFBP and P4 concentrations in FF. These data highlight the importance of vascularity, angiogenic factors, and IGFBP in the health of the dominant follicle in heifers, and indicate that the FF concentrations of E, VEGF, IGFBP, and P4, and the E:P4 ratio can be used as markers of dominant follicles.

O-63: Angiogenesis factors in human ovarian follicular development: The presence of vascular endothelial growth factor and placental growth factor in follicular fluid

It has been demonstrated that angiogenesis plays an important role in normal cyclical ovarian function. In particular, follicular growth is dependent upon the proliferation of new capillary vessels, and, it has been proposed that the process of selection of a dominant follicle is likely to be associated with angiogenesis. Vascular endothelial growth factor (VEGF) and its homolog the placental growth factor (PlGF) are vascular cell-specific mitogen factors with a key function in angiogenesis. Their role in human follicular growth and oocyte maturation however, is less well-understood and the presence of PlGF in human follicles has not been previously demonstrated. This study sought to determine the content of both VEGF and PlGF within the follicular fluid (FF) of large and small follicles of women undergoing in vitro fertilization (IVF) and to relate their content to patient age A prospective laboratory-based study at an academic center. Women between the ages of 25 - 43 years old undergoing IVF were enrolled under IRB approval during the time of oocyte retrieval. Follicles were measured by transvaginal ultrasound in 2 dimensions prior to aspiration. FF from lead (>14mm) and small (< 12mm) follicles was collected by transvaginal aspiration. Cellular components and immunoglobulins were removed by centrifugation and protein A/G chromatography. VEGF and PlGF concentration in FF were then analyzed via enzyme-linked immunosorbent assay (ELISA) on lead and small follicles from young (</= 34 years) and old (>/= 40 years) samples and confirmed by western blot with specific antibodies. Results were analyzed with students’t test and statistical significance was set at a p value of < 0.05. The results of the Western Blot analysis showed the presence of both VEGF and PlGF within the FF. The mean concentration of PlGF in lead follicles was 42.41 pg/ml and the mean concentration in small follicles was 55.91 pg/ml (p= 0.10). The mean concentration of VEGF in lead follicles was 5,745.76 pg/ml whereas the mean concentration in small follicles was 3,485.70 pg/ml (p<0.01). The concentration of VEGF and PlGF did not vary with respect to age. This study demonstrated a significantly increased concentration of VEGF in the follicular fluid of lead follicles compared with smaller follicles irrespective of age, lending proof to the idea that angiogenesis is an important part of lead human follicle development. The trend of our results for PlGF supports findings suggesting that PlGF may act as a natural VEGF antagonist. Additionally, our western blot and ELISA results are the first demonstration of the presence of PlGF in human FF. Overall, this study demonstrates that angiogenesis factors are important in follicular development in women undergoing IVF.

Gonadotropin-releasing hormone antagonists increase follicular fluid insulin-like growth factor-I and vascular endothelial growth factor during ovarian stimulation cycles

The aim of the present study was to investigate the effect of gonadotropin-releasing hormone (GnRH) antagonists (GnRH-ant) on follicular fluid (FF) insulin-like growth factor-I (IGF-I) and FF vascular endothelial growth factor (VEGF) levels. Sixty women undergoing assisted reproduction were randomized and assigned to two different GnRH analog regimens: GnRH agonist (GnRH-a) and GnRH-ant. FF VEGF and FF IGF-I concentrations were significantly increased in the patients treated with GnRH-ant (p < 0.001). In the same patients we observed a statistically significant reduction in serum luteinizing hormone (LH) and estradiol (E2) levels (p < 0.001 and p < 0.05, respectively), FF E2 and FF androstenedione levels (p < 0.05 and p < 0.001, respectively), as well as a reduction in the number of pregnancies although this was not statistically significant. In the GnRH-ant group, FF VEGF levels were positively correlated with FF IGF-I levels, and both were negatively correlated with serum LH levels. The increase in FF IGF-I and FF VEGF levels in women treated with GnRH-ant could be explained by a deleterious follicular environment in response to profound suppression of LH and E2 levels.

Vascular endothelial growth factor and basic fibroblast growth factor in polycystic ovary syndrome during controlled ovarian hyperstimulation

Background. Polycystic ovary syndrome (PCOS) is a common endocrine disorder that causes anovulation and consequent subfertility. It is well established that increased ovarian mass, supported by new blood vessel proliferation in stroma and theca, is a key feature of PCOS. Recent studies suggest a role for angiogenetic factors in this phenomenon.Aim. To evaluate of levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in serum and follicular fluid of PCOS patients during a controlled ovarian hyperstimulation.Methods. In 52 patients undergoing in vitro fertilization treatments, 26 PCOS patients and 26 controls, serum VEGF and bFGF levels were assessed before starting administration of follicle-stimulating hormone (FSH) (day 0), on the day of administration of human chorionic gonadotropin (hCG) and on the day of oocyte retrieval. Follicular fluid levels of the two growth factors were detected on the day of oocyte retrieval.Results. PCOS patients showed higher serum VEGF levels than controls before starting FSH administration, on the day of hCG administration and on the day of oocyte retrieval. Serum VEGF levels showed a rise after hCG administration only in the PCOS patients. In addition, serum bFGF levels were higher in PCOS patients than in controls on the day of hCG administration and the day of oocyte retrieval. Furthermore, on the day of hCG administration, serum bFGF levels were directly correlated to the amount of FSH previously administered (p < 0.0001). In follicular fluid, higher VEGF and bFGF levels were found in PCOS patients than in controls. Furthermore, follicular-fluid bFGF concentrations were inversely correlated with the percentage of mature oocytes collected (p < 0.05).Conclusions. The present study revealed elevated levels of VEGF and bFGF in serum and follicular fluid in PCOS patients compared with controls. bFGF seems to be an FSH-dependent growth factor and its levels in follicular fluid are inversely correlated with the percentage of mature oocytes collected.

Relationship Between Etiology of Sterility of Patients Undergoing Controlled Ovarian Hyperstimulation (COH) and Serum, Follicular Fluid Concentration of Leukaemia Inhibitory Factor (LIF), Vascular Endothelial Growth Factor (VEGF), Nitic Oxide (NO) and Their Influence on ICSI Outcome

The aim of this study was to determine the concentration of LIF, VEGF and NO concentration in serum and follicular fluid of tubar, endometriosis and male factor infertility patients who undergoing COH for intracytoplasmatic sperm injection (ICSI) and to find out (i) the difference between the FF and serum concentration, (ii) the relationship between the etiology of sterility and these growth factors concentrations, (iii) the influence of these investigated growth factor on ICSI outcome. Prospective study. 75 women undergoing ICSI therapy were included in this study. Tubar sterility (G.1=17), endometriosis (G.2 = 15) and male factor infertility (G.3. 43). After pituitary down regulation using Gn-RH a, the women underwent COH either with rFSH or HMG/FSH. Serum and FF were collected at the time of oocytes retrieval and were frozen at -80°C until the measurement of LIF; VEGF and NO concentration. LIF and VEGF concentration was determined by using enzyme-linked immunosorbent assay (ELISA). Whereas, the NO was measured using the Griess reaction as an indirect assessment of NO activity. LIF concentrations in FF of patients with tubal sterility, endometriosis, male factor sterility were (43.20±12.10, 43.6±18.9 and 50.0±23.70 pg/ml respectively) and the corresponding values in serum were (1.60±0.90; 1.50±0.96 and 2.30±2.4 pg/ml). VEGF concentrations in FF of the first, second and third groups were (3247.8±931.9; 3546.3±893.40 and 3358.1±894.3 pg/ml) and in serum as follows (375.3±180.5; 354.3±160.5 and 374.8±182.4). NO concentrations were at the same level in FF and serum in the first group (42.3±9.50 vs. 39.190± 15.52, p=0, 26; 41.5±8.99vs.36.3±13.6 p= 0.18) in the second group and 40.1±12.5 vs. 35.96±18.9 p= 0.059 μmol/l in the third group. LIF concentration in FF was 43.2±12.1 and in 1.6±0.9 pg/ml in serum (p<0.001). VEGF in FF was 3247.8±931.9 and in serum 375.3±180.5 pg/m (p<0.001). NO concentrations in FF and serum were similar (42.3±9.5 vs. 39.19.52μmol/l; p=0.26). The main number of retrieved, fertilized and transferred oocytes in the first group was (8.2±5.3; 4.7±3.7 and 2.0±0.8) the corresponding value in the second group was (5.0±3.6; 3.7±3.1 and 1.9±0.9) and in the third group (5.9±4.1; 3.4±2.7 and 1.7±0.9). No significance difference was shown between the groups in respect of the age of the patient (p=0.31), the main number of retrieved (p=0.132), fertilized (p=0.29) and transferred oocytes (p=0.73). Moreover, the fertilization rate, in all three investigated groups (64.0±2.9%, 81.0±3.1% and 60.0±4.5%, p=0.42), cleavage (60. 0±3.1%; 69.0±2.9% and 65.0±2.8%; p=0.73) and ongoing pregnancy rates (17.6%; 46.7%; 20.9% p=0.19) were similar. The concentration of NO and LIF in FF correlate negatively with fertilization (-0.012; -0.218) and cleavage rate (-0.10; -0.067) of the oocytes. The concentration of LIF, VEGF and NO in the investigated groups was similar. LIF and VEGF concentration was significantly higher in FF in comparison to serum, which indicated that these growth factors synthesised in the ovary and play an important role in ovarian physiology. Whereas, NO concentration was at the same levels in FF and serum. Besides, a negative correlation was shown between NO concentration in FF, serum and fertilization and cleavage rates of the fertilized oocytes.