Feskov O. M., Zhylkova Ye. S., Chumakova N. O., Feskova А. O., Yehunkova O. V.

REACTIVE OXYGEN SPECIES IN EJACULATE AFFECT EMBRYO DEVELOPMENT IN VITRO WHEN ASSISTED REPRODUCTIVE TECHNOLOGIES ARE USED

---

Show/Download

About the author:

Feskov O. M., Zhylkova Ye. S., Chumakova N. O., Feskova А. O., Yehunkova O. V.

Heading:

CLINICAL AND EXPERIMENTAL MEDICINE

Type of article:

Scientific article

Annotation:

Reactive oxygen species are a necessary condition for the maturation, existence and ability of spermatozoa to fer tilize the oocyte. The excess of reactive oxygen species in semen can initiate pathological changes in sperm, causing oxidative damage to cell membranes, proteins and DNA. Currently, several scientific studies indicate that high levels of reactive oxygen species in the ejaculate are one of the causes of low male reproductive function. On the other hand, the measurement of reactive oxygen species is hampered by the lack of standardization, including the choice of controls and sample selection. The published data about the effects of oxidative stress on standard fertility sperm parameters (sperm motility, concentration and morphology) and on the integrity of sperm DNA in the ejaculate are controversial. The aim of the present work was to investigate the effect of reactive oxygen species in ejaculate on standard mi croscopic sperm parameters and on the early embryo development in vitro in couples with male factor of infertility. The development of the embryos from 26 married couples with combined factors of infertility was analyzed. Microscopic analysis of ejaculate was performed according to WHO recommendations from 2021. The level of reac tive oxygen species (ROS) in ejaculate was analyzed using Oxisperm kits (Halotech DNA, Spain). Donor oocytes were fertilized by the method of intracytoplasmic sperm injection (ICSI). Embryos that reached the blastocyst stage were evaluated by morphological characteristics according to the criteria of D. Gardner. Preimplantation genetic testing of blastocysts for aneuploidy was performed using the next-generation sequencing method. Statistical hypotheses were tested using the t, chi-square and rs criteria at significance levels of 0.05 and 0.01. The negative influence of the excess of reactive oxygen species in semen on the early embryo development in vitro is proved. The ROS-level in ejaculate in infertile patients are significantly higher comparing with the ROS-value for men with normal microscopic sperm parameters (2.4±0.8 vs. 1.6±0.6 р<0.05). A significant negative correlation was found between sperm motility and paternal age (rs=-0.54, p=0.01795). Total blastocyst formation rate for embryos in vitro was significantly lower in infertile patients, comparing with the mentioned index for embryos obtained from donor gametes (50.2% vs. 76.0%, df=1, χ2fact.=15.3173, p=0.000157). The strong negative correlation between the ROS-level and both total blastocysts formation rate and euploid blastocyst formation rate was demonstrated in in fertile patients (rs=-0.66, p=0.00247 and rs=-0.65, p=0.04034, respectively).

Tags:

Bibliography:

1. Mazzilli R, Rucci C, Vaiarelli A, Cimadomo D, Ubaldi FM, Foresta C, et al. Male factor infertility and assisted reproductive technologies: indications, minimum access criteria and outcomes. J Endocrinol Invest. 2023;46(6):1079-1085. DOI: 10.1007/s40618-022-02000-4.
2. Mannucci A, Argento FR, Fini E, Coccia ME, Taddei N, Becatti M, et al. The Impact of Oxidative Stress in Male Infertility. Front Mol Biosci. 2022;8:799294. DOI: 10.3389/fmolb.2021.799294.
3. Wagner H, Cheng J, Ko Edmund Y. Role of reactive oxygen species in male infertility: An updated review of literature. Arab J Urol. 2018;16(1):35-43. DOI: 10.1016/j.aju.2017.11.001.
4. Parekattil SJ, Esteves SC, Agarwal A, editors. Male infertility. Switzerland: Springer; 2020. Chapter, Apoptosis and male infertility; p. 479-486.
5. Bisht S, Dada R. Oxidative stress: Major executioner in disease pathology, role in sperm DNA damage and preventative strategies. Frontiers in Bioscience (Schol Ed). 2017;9:420-447.
6. O’Flaherty C. Reactive Oxygen Species and Male Fertility. Antioxidants. 2020;9(4):287. DOI: 10.3390/antiox9040287.
7. Liu J, Zhu K, Xu S, Tu W, Lin X, Su Y, et al. Double-edged sword: effects of human sperm reactive oxygen species on embryo development in IVF cycles. Reprod Biol Endocrinol. 2023 4;21(1):1. DOI: 10.1186/s12958-022-01053-7.
8. Kuroda S, Takeshima T, Takeshima K, Usui K, Yasuda K, Sanjo H, et al. Early and late paternal effects of reactive oxygen species in semen on embryo development after intracytoplasmic sperm injection. Systems Biology in Reproductive Medicine. 2020;66(2):122-128. DOI: 10.1080/19396368.2020. 1720865.
9. Burruel V, Klooster K, Barker C. Abnormal Early Cleavage Events Predict Early Embryo Demise: Sperm Oxidative Stress and Early Abnor mal Cleavage. Sci Rep. 2014;4:6598. DOI: 10.1038/srep06598.
10. Yunghua F, Jianying H, Jie W, Lin Q, Wen O, Ximeng A, et al. Effect of sperm swim − up vs density gradient centrifugation on embryo aneuploidy detected by non-invasive chromosomal screening in conventional IVF. European Journal of Obstetrics & Gynecology and Re productive Biology. 2025;312:114545. DOI: 10.1016/j.ejogrb.2025.114545.
11. Chung E, Atmoko W, Saleh R, Shah R, Agarwal A. Sixth edition of the World Health Organization laboratory manual of semen analysis:Up dates and essential take away for busy clinicians. Arab Journal of Urology. 2024;22(2):71-74. DOI: 10.1080/20905998.2023.2298048.
12. Castleton PE, Deluao JC, Sharkey DJ, McPherson NO. Measuring Reactive Oxygen Species in Semen for Male Preconception Care: A Scientist Perspective. Antioxidants (Basel). 2022;11(2):264. DOI: 10.3390/antiox110 20264.
13. Gruber I, Klein M. Embryo culture media for human IVF: which possibilities exist? J Turk Ger Gynecol Assoc. 2011;12(2):110-117. DOI: 10.5152/jtgga.2011.25.
14. Gardner DK, Lane M, Stevens J. Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril. 2000;73:1155-1158. DOI: 10.1016/S0015-0282(00)00518-5.
15. Doroftei B, Ilie O, Anton N, Armeanu T, Ilea C. A Mini-Review Regarding the Clinical Outcomes of In Vitro Fertilization (IVF) Follow ing Pre-Implantation Genetic Testing (PGT)-Next Generation Sequencing (NGS) Approach. Diagnostics (Basel). 2022;12(8):1911. DOI: 10.3390/diagnostics 12081911.
16. Petrie A, Sabin C. Medical Statistics at a Glance. Winnipeg: Blackwell Science; 2000. 157 р.
17. Piccolomini MM, Bonetti TC, Motta E, Serafini PC, Alegretti JR. How general semen quality influences the blastocyst formation rate: Anal ysis of 4205 IVF cycles. JBRA Assist Reprod. 2018;22(2):89-94. DOI: 10.5935/1518-0557.20180022.
18. Durairajanayagam D, Singh D, Agarwal A, Henkel R. Causes and consequences of sperm mitochondrial dysfunction. Andrologia. 2021;53:e13666.
19. Kumaresan A, Das Gupta M, Datta TK, Morrell JM. Sperm DNA Integrity and Male Fertility in Farm animals: a review. Front Vet Sci. 2020;7:321.

Publication of the article:

«Bulletin of problems biology and medicine», 2026 Issue 1, 180, 380-388 pages, index UDC 573.6:577.21-076

DOI:

10.29254/2077-4214-2026-1-180-380-388