Abstract

Abstract Study question Do seminal oxidation-reduction potential (ORP) and sperm DNA fragmentation (SDF) predict ICSI cycle outcomes? Summary answer Both SDF and seminal ORP have strong prognostic value in predicting good fertilization (≥80%), blastocyst development (≥60%), clinical pregnancy and live birth. What is known already Seminal oxidative stress (OS) has been found to play an important role in various aetiologies of male infertility and is induced by the imbalance between reactive oxygen species (ROS) and antioxidants. OS is a major cause of sperm DNA fragmentation (SDF). It is well established that high levels of seminal OS and SDF are negatively associated with ART outcomes. Previous studies have associated high levels of SDF with an increase risk of miscarriage and a decrease in live birth rate. Study design, size, duration This prospective study included 144 couples who had a fresh autologous ICSI cycle between June 2018 and December 2020. There was no restriction to patients with severe male factor infertility. All cryopreservation and third party assisted reproductive techniques were excluded. Participants/materials, setting, methods This multi-centre study was performed at three ART Clinics in Cape Town, South Africa. All couples included in the study had fresh autologous ICSI cycles with freshly ejaculated semen. Seminal ORP testing was performed using the MiOXSYS system and SDF by means of a fluorescence microscopic TUNEL assay on freshly ejaculated semen samples that were used for ICSI. The ART outcomes evaluated were fertilization rate, blastocyst development rate, clinical pregnancy rate and live birth rate. Main results and the role of chance The study shows that seminal ORP significantly negatively correlates with fertilization rate (r=-0.267; P = 0.0012), blastocyst development rate (r=-0.389; P < 0.0001), pregnancy (r=-0.296; P = 0.0003) and live birth (r=-0.353; P < 0.0001). ROC curve analysis shows significant predictive power for ORP for fertilization (≥80%; AUC=0.652; P = 0.0012), blastocyst development rate (≥60%; AUC=0.762; P < 0.0001), pregnancy (AUC=0.677; P = 0.0002) and live birth (AUC=0.729; P < 0.0001). Comparable results were obtained for SDF. An average cut-off value for ORP of 0.55 mV/106 sperm/mL was calculated. The AUCs between the SDF and ORP results for all reproductive outcome parameters did not differ. For all reproductive outcome parameters, normal sperm morphology shows the lowest predictive power. ORP with male age as confounding factor had significant effects on odds ratios (OR) of fertilization (OR = 0.1446; CI: 0.048-0.430; P = 0.0001) and blastocyst development (OR = 0.2468; CI: 0.095-0.635; P = 0.0029), while it was not significant for pregnancy and life birth. Multivariate logistic regression with fertilization, blastocyst development, pregnancy and live birth as dependent parameters and high/low seminal ORP, primary and secondary infertility, unexplained infertility, polycystic ovaries, endometriosis, tubal factor myomectomy, ovarian insufficiency and advanced maternal age as independent variables showed overall model fits (P < 0.0002) with a significant (P < 0.0084) influence of ORP on all reproductive outcome parameters. Limitations, reasons for caution Since only ICSI patients were included in this study, this might have contributed to the fact the cut-off value of ORP is lower than the published cut-off. A further limitation is the use of a fluorescence microscopic TUNEL assay which drastically limited the number of sperm evaluated. Wider implications of the findings This is the first study showing predictive value of seminal ORP for reproductive outcome including pregnancy and live birth. The method is easy and much quicker than current methods for the determination of SDF. Hence, it can be used a point of care method. Trial registration number Not applicable

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