Effect Of Unsurgically Treated Ovarian Endometrioma On In Vitro Fertilization Outcome And Ferroptosis Status Of Follicular Granulosa Cells

BackgroundThe effects of ovarian endometrioma(OMA)on follicles,oocyte quality,embryo euploidy rate,endometrial receptivity and pregnancy outcome of in vitro fertilization(IVF)are still controversial,and ferroptosis is one of the potential mechanisms.The aim of this study is to analyze the effects of unsurgically treated OMA on IVF outcomes from the five aspects and explore the role of ferroptosis on follicles.Objectives1.To investigate whether OMA affects the cumulative pregnancy outcome and efficiency of multiple IVF in patients with decreased ovarian reserve(DOR);2.To investigate whether OMA affects the embryo euploidy rate and pregnancy outcome of PGT cycle in DOR patients;3.To investigate whether OMA increases the incidence of chronic endometritis(CE)and implantation window deviation in patients with implantation failure;4.To detect ferroptosis markers in follicular fluid of OMA patients and their relationship with oocyte quality.5.To investigate the effect of OMA on ferroptosis of follicular granulosa cells.Methods1.A retrospective analysis was performed on 1079 IVF cycles of 493 patients(90 in OMA group and 403 in non-OMA group)with DOR,and the pregnancy outcomes and time to live birth were compared between the two groups.2.A total of 123 PGT cycles of 100 DOR patients(20 patients in OMA group and 80 patients in non-OMA group,1:4 matched according to age,PGT type and pregnancy history)were retrospectively analyzed,and the euploidy rate of blastocyst and pregnancy outcome were compared between the two groups.3.Retrospective analysis:a total of 368 patients(40 patients in OMA group and 328 patients in non-OMA group)after more than one implantation failure who underwent CE detection were enrolled.The incidence of CE and pregnancy outcomes after treatment were compared between the two groups.Endometrial receptivity analysis(ERA)was performed on 30 patients(10 patients in OMA-ERA group and 20 patients in non-OMA-ERA group)with repeated implantation failure.The proportion of ERA type,implantation window deviation rate and pregnancy outcome were compared between the two groups.4.The levels of ferroptosis markers Fe2+,GSH(glutathione)and malondialdehyde(MDA)in the follicular fluid of the first follicle of 40 patients(matched by age and pregnancy history,20 patients in the OMA group and 20 patients in the non-OMA group)were detected.The differences of the three ferroptosis markers in the two groups were compared,and their correlation with oocyte and embryo quality was analyzed.5.Ferroptosis markers,mitochondrial morphology,GSSG,MDA,ROS,Beclin-1,SLC7A11 and GPX4 levels in follicular fluid of the three groups(OMA side,OMA contralateral and non-OMA group,8 cases in each group)were detected and compared.Results1.There was no significant difference in the implantation rate,live birth rate per oocyte retrieval cycle and per transfer cycle,cumulative live birth rate per patient and per patient with high-quality embryos between two groups,and there was no significant difference in the time to live birth(P>0.05).2.There were no significant differences between OMA group and non-OMA group in euploidy rate,implantation rate o,clinical pregnancy rate,miscarriage rate and live birth rate(P>0.05).3.The detection rate of CE was similar between OMA group and non-OMA group,and the cumulative pregnancy rate and live birth rate after treatment were similar in both groups.There were no significant differences between the OMA-ERA group and the non-OMA-ERA group in the rate of implantation window deviation and pregnancy rate of the subsequent embryo transfer(P>0.05).4.The oocyte retrieval rate of the first follicle and the total oocyte retrieval rate in OMA group were significantly lower than those in non-OMA group,the levels of Fe2+and MDA in follicular fluid in OMA group were significantly higher than those in nonOMA group,and the level of GSH in OMA group was lower than that in non-OMA group,and the differences were statistically significant(P<0.05).The level of GSH in follicular fluid without oocytes was significantly lower than that in follicular fluid with oocytes,and GSH was positively correlated with the rate of D3 high-quality embryo in OMA group(P<0.05).5.The levels of ferroptosis markers in follicular fluid,mitochondrial morphology,GSSG,MDA,ROS and pathway proteins Beclin-1,SLC7A11 and GPX4 in granulosa cells of the three groups(OMA side,OMA contralateral and non-OMA group,8 cases in each group)were detected and compared.ConclusionsUnsurgically treated OMA did not affect the cumulative pregnancy outcome of multiple IVF in DOR patients and did not affect the time to live birth.OMA did not affect the rate of blastocyst euploidy and pregnancy outcome in DOR patients.In patients with implantation failure,the incidence of CE and implant window deviation in OMA patients were similar to those in non-OMA women.Therefore,for patients with DOR with 3-5 cm OMA,it is not recommended to perform surgical treatment only to improve pregnancy outcome before IVF.Unilateral OMA may induce ferroptosis in the adjacent follicle granulosa cells,resulting in oxidative stress and thus affecting the local oocyte quality.