Correlations between Follicular fluid PlGF and IVF/ICSI outcomes among polycystic ovary syndrome women and normo-ovulatory women using different controlled hyperstimulation protocols
Authors: Kadoura Sally , Alhalabi Marwan , Nattouf Abdul Hakim 
Published: April 2022

Abstract
Background
Gonadotropin-releasing hormone (GnRH) analogues are commonly used in clinical practice to prevent premature luteinizing hormone (LH) surge during In-Vitro Fertilization/ Intra-Cytoplasmic Sperm Injection (IVF/ICSI) cycles. However, based on our recent work, the follicular fluid levels of the Placental growth factor (FF PlGF), the novel angiogenic factor, differ significantly between GnRH agonist and GnRH antagonist protocols. Thus, we conducted this study to compare the IVF/ICSI outcomes and their correlations with FF PlGF levels in polycystic ovary syndrome (PCOS) women and normo-ovulatory women during different controlled hyperstimulation protocols.
Methods
The current study is a re-analysis of our previous work. The data were adopted from two prospective trials that were conducted on women who were referred to the Assisted Reproductive Unit of Orient Hospital, Damascus, Syrian Arab Republic, from December 2019 to August 2021. A total of 75 PCOS women (Rotterdam criteria) (GnRH agonist group, PCOSA, n = 53; GnRH antagonist group, PCOSAnta, n = 22) and 83 normo-ovulatory women (GnRH agonist group, ControlA, n = 50; GnRH antagonist group, ControlAnta, n = 33) were included. Follicular fluid samples were collected on the retrieval day, and the FF levels of Placental growth factor (PlGF) and Anti-Müllerian hormone (AMH) were measured using ELISA Kits. Before being subjected to ICSI, the mature oocytes from both groups were morphologically assessed under an inverted microscope at 400x magnification. In addition, the embryological and clinical IVF/ICSI outcomes were detected. Spearman rank correlation coefficients were computed to assess the correlation among the studied parameters. The area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the accuracy of follicular fluid PlGF levels in predicting pregnancy rates.
Results
There were not any significant differences between PCOSA and ControlA groups or PCOSAnta and ControlAnta groups at baseline characteristics. Although PCOS women were stimulated using lower doses of gonadotropins (PCOSA = 1868.40 ± 668.29 IUs vs ControlA = 2523.00 ± 1034.11 IUs; P value < 0.001), (PCOSAnta = 1779.55 ± 702.87 IUs vs ControlAnta = 2468.18 ± 879.53 IUs; P value = 0.003), the number of retrieved oocytes, MII oocytes, MI oocytes, and immature oocytes were significantly higher in the PCOS group compared to the Control group during the GnRH agonist protocol, but not GnRH antagonist one. Nevertheless, OSI values were significantly higher in PCOS groups independently of the protocol used (PCOSA = 11.83 ± 6.98 Oocyte/IU vs ControlA = 7.48 ± 4.75 Oocyte/IU; P value < 0.001), (PCOSAnta = 11.46 ± 8.99 Oocyte/IU vs ControlAnta = 7.37 ± 4.87 Oocyte/IU; P value = 0.042). On the other hand, there were no significant differences between PCOS and controls in maturation rate, fertilization rate, highquality embryos rate, cleavage rate, implantation rate or oocytes morphology independently on the protocol used. During both protocols, the FF AMH levels were significantly higher in the PCOS groups compared to the Control ones, while the FF PlGF levels were similar between them. Regarding correlations between FF PlGF and IVF/ICSI outcomes, FF PlGF levels were negatively correlated with age and total gonadotropins dose and positively correlated with OSI in the PCOSAnta, ControlA, and ControlAnta groups, but not in the PCOSA group. Moreover, FF PlGF levels positively correlated with the number of MII oocytes in the PCOSAnta group and the number of retrieved oocytes in the ControlA group. Nevertheless, no significant differences were noted in FF PlGF levels between pregnant and non-pregnant women in any of the studied groups, which also was confirmed by the Receiver Operating Characteristic (ROC) Curve analysis.
Conclusions
Although PCOS exaggerates ovarian response to stimulation irrespective of the protocol used, it does not have a detrimental impact on oocytes morphology, quality, or competence. In addition, FF PlGF levels could be a marker to the ovarian response other than a predictor of pregnancy achievement during IVF/ICSI cycles independent of the PCOS pathology.
Study registration:
The data were adopted from two prospective clinical trials that were registered on the clinicaltrials.gov site by registration numbers NCT04727671 and NCT04724343.