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Comparing reactive oxygen species and DNA fragmentation in semen samples of unexplained infertile and healthy fertile men



Male factor infertility has increased to more than 40% during the last decade. About 30% of these couples are diagnosed with unexplained infertility. In fact, reactive oxygen species (ROS), especially superoxide anion (O2 ·) and hydrogen peroxide (H2O2), play a crucial role in regulation of physiological and pathological processes in spermatozoa. Moreover, since the diagnosis of unexplained infertility just through semen analysis is a matter of much controversy; we aimed to evaluate the levels of ROS and sperm DNA fragmentation in the semen samples of unexplained infertile and fertile control couples.


The semen samples of 28 unexplained infertile couples and 30 fertile control couples were analyzed according to WHO criteria. The intracellular levels of H2O2 and O2 · were detected by flow cytometry with 2′,7′-Dichlorodihydrofluorescin diacetate and Dihydroethidium, respectively, and DNA fragmentation was evaluated by sperm chromatin dispersion test.


In unexplained infertile group, sperm motility and normal morphology were significantly lower than the control. The levels of sperm H2O2, O2 ·, and DNA fragmentation were significantly higher in unexplained infertile men compared to fertile. Moreover, a positive correlation was found between the level of H2O2 and sperm DNA fragmentation in the unexplained infertile group. Besides, reduced sperm motility in the unexplained infertile group was significantly correlated with elevated levels of ROS.


The higher levels of intracellular ROS and DNA fragmentation in the semen samples of unexplained infertile couples and their causes might be considered as an important factor related to diagnosis and treatment of the unexplained infertile couples.

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We wish to thank Miss Elaheh Aboutorabi for her skilful technical assistance.


This study was financed by Reproductive Health Research Centre (grant no. 910439-19441).

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Correspondence to Saeid Amanpour.

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Zandieh, Vatannejad, Doosti, Zabihzadeh, Haddadi, Bajelan, Rashidi have no conflict of interest. Amanpour has received research grants from Reproductive Health Research Centre.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

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Zahra Zandieh and Akram Vatannejad are equal first author.

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Zandieh, Z., Vatannejad, A., Doosti, M. et al. Comparing reactive oxygen species and DNA fragmentation in semen samples of unexplained infertile and healthy fertile men. Ir J Med Sci 187, 657–662 (2018).

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  • DNA damage
  • Human spermatozoa
  • Reactive oxygen species
  • Unexplained infertility