A Comparison Between Two Assays for Measuring Seminal Oxidative Stress and their Relationship with Sperm DNA Fragmentation and Semen Parameters

Sheryl T Homa, Anna M Vassiliou, Jesse Stone, Aideen P Killeen, Andrew Dawkins, Jingyi Xie, Farley Gould, Jonathan W A Ramsay, Sheryl T Homa, Anna M Vassiliou, Jesse Stone, Aideen P Killeen, Andrew Dawkins, Jingyi Xie, Farley Gould, Jonathan W A Ramsay

Abstract

Oxidative stress (OS) is a significant cause of DNA fragmentation and is associated with poor embryo development and recurrent miscarriage. The aim of this study was to compare two different methods for assessing seminal OS and their ability to predict sperm DNA fragmentation and abnormal semen parameters. Semen samples were collected from 520 men attending for routine diagnostic testing following informed consent. Oxidative stress was assessed using either a chemiluminescence assay to measure reactive oxygen species (ROS) or an electrochemical assay to measure oxidation reduction potential (sORP). Sperm DNA fragmentation (DFI) and sperm with immature chromatin (HDS) were assessed using sperm chromatin structure assay (SCSA). Semen analysis was performed according to WHO 2010 guidelines. Reactive oxygen species sORP and DFI are negatively correlated with sperm motility (p = 0.0012, 0.0002, <0.0001 respectively) and vitality (p < 0.0001, 0.019, <0.0001 respectively). The correlation was stronger for sORP than ROS. Reactive oxygen species (p < 0.0001), sORP (p < 0.0001), DFI (p < 0.0089) and HDS (p < 0.0001) were significantly elevated in samples with abnormal semen parameters, compared to those with normal parameters. Samples with polymorphonuclear leukocytes (PMN) have excessive ROS levels compared to those without (p < 0.0001), but sORP and DFI in this group are not significantly increased. DNA fragmentation was significantly elevated in samples with OS measured by ROS (p = 0.0052) or sORP (p = 0.004). The results demonstrate the multi-dimensional nature of oxidative stress and that neither assay can be used alone in the diagnosis of OS, especially in cases of leukocytospermia.

Keywords: DNA fragmentation; DNA oxidation; chromatin; male infertility; oxidative stress; reactive oxygen species; spermatogenesis.

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Box and whisker plots for seminal oxidative stress levels in different patient groups showing the median and interquartile ranges. Normal—normal semen parameters; Abnormal—abnormal parameters with a) chemiluminescence (ROS) or (b) oxidation reduction potential (sORP) Lower whisker = 10th percentile; upper whisker = 90th percentile. Dots indicate values outside the range. Data are shown on a logarithmic scale.
Figure 2
Figure 2
Box and whisker plots for sperm genetic integrity in different patient groups showing the median and interquartile ranges. Normal—normal semen parameters; Abnormal—abnormal parameters with a) DNA fragmentation (DFI) (b) immature chromatin (HDS). Lower whisker = 10th percentile; upper whisker = 90th percentile. Dots indicate values outside the range. Data are shown on a logarithmic scale.

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