Archives of Gynecology and Obstetrics

, Volume 300, Issue 1, pp 207–215 | Cite as

The impact of sperm DNA fragmentation on ICSI outcome in cases of donated oocytes

  • Sevastiani AntonouliEmail author
  • Achilleas Papatheodorou
  • Yannis Panagiotidis
  • Stamatios Petousis
  • Nikos Prapas
  • Stefania Annarita Nottola
  • Maria Grazia Palmerini
  • Guido Macchiarelli
  • Yannis Prapas
Gynecologic Endocrinology and Reproductive Medicine



The aim of this study is to evaluate the sperm DNA fragmentation index (DFI) in oocyte donation cycles and correlate it with the sperm parameters, the male characteristics, the embryo quality and the outcome of intracytoplasmic sperm injection (ICSI).


A total of 150 couples participating in an oocyte donation program were included in the study. Sperm samples were assessed by conventional sperm analysis. DFI was evaluated using the Halosperm kit, a sperm chromatin dispersion test (SCD).


The relations between DNA damage and epidemiological male factors (age, height, weight), standard semen parameters (concentration, total and forward motility, and morphology), and embryological and clinical parameters (fertilization rate, total blastocyst number, number of good quality blastocyst, clinical pregnancy) were analyzed. DFI was positively correlated with advanced male age (r = 0.23, p < 0.05) and negatively correlated with total sperm and forward motility (r = − 0.29, r = − 0.27, respectively; p < 0.05). DFI was not significantly correlated with pregnancy outcome in oocyte donation cycles (r = − 0.05, p > 0.05). When good quality blastocysts were chosen, a trend toward the development of good quality embryos was detected in the presence of a low DFI (r = − 0.20, p = 0.08).


DFI does not significantly affect the outcome of ICSI in oocyte donation cycles. Even in cases of advanced paternal age that a high DFI resulted sperm DNA fragmentation seems not to adversely affect the final outcome.


DNA fragmentation SCD test Infertility ICSI Oocyte donation 



Preliminary data regarding this study were presented as poster at the 10th European Congress of Andrology, October 2018 in Budapest, Hungary.

Author contribution

SA: Data collection or management, data analysis, manuscript writing. AP: Protocol/project development, data analysis, manuscript writing/editing. YPa: Protocol/project development, data analysis. SP: Data management, data analysis. NP: Data management, data analysis, manuscript editing. SAN: Data analysis, manuscript editing. MGP: Data collection or management. GM: Data analysis, manuscript editing. YPr: Protocol/project development, data management, data analysis, manuscript writing/editing.


This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

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. The Institutional Review Board of Iakentro Advanced Medical Center/ethics committee of Thessaloniki, Greece approved the study at 2/9/2013, Ref. Number 9/2013.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. 1.
    Schulte RT, Ohl DA, Sigman M, Smith GD (2010) Sperm DNA damage in male infertility: etiologies, assays, and outcomes. J Assist Reprod Genet 27:3–12. CrossRefGoogle Scholar
  2. 2.
    Sakkas D, Alvarez JG (2010) Sperm DNA fragmentation: mechanisms of origin, impact on reproductive outcome, and analysis. Fertil Steril 93:1027–1036. CrossRefGoogle Scholar
  3. 3.
    Avendaño C, Franchi A, Duran H, Oehninger S (2010) DNA fragmentation of normal spermatozoa negatively impacts embryo quality and intracytoplasmic sperm injection outcome. Fertil Steril 94:549–557. CrossRefGoogle Scholar
  4. 4.
    Li Z, Wang L, Cai J, Huang H (2006) Correlation of sperm DNA damage with IVF and ICSI outcomes: a systematic review and meta-analysis. J Assist Reprod Genet 23:367–376. CrossRefGoogle Scholar
  5. 5.
    Aitken RJ, Gordon E, Harkiss D, Twigg JP, Milne P, Jennings Z, Irvine DS (1998) Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod 59:1037–1046CrossRefGoogle Scholar
  6. 6.
    Taherzadeh S, Khalili MA, Agha-Rahimi A, Anbari F, Ghazali S, Macchiarelli G (2017) Vitrification increased vacuolization of human spermatozoa: application of MSOME technology. J Reprod Infertil 18:225–230Google Scholar
  7. 7.
    Nottola SA, Albani E, Coticchio G, Palmerini MG, Lorenzo C, Scaravelli G, Borini A, Levi-Setti PE, Macchiarelli G (2016) Freeze/thaw stress induces organelle remodeling and membrane recycling in cryopreserved human mature oocytes. J Assist Reprod Genet 33:1559–1570. CrossRefGoogle Scholar
  8. 8.
    Palmerini MG, Antinori M, Maione M, Cerusico F, Versaci C, Nottola SA, Macchiarelli G, Khalili MA, Antinori S (2014) Ultrastructure of immature and mature human oocytes after cryotop vitrification. J Reprod Dev 60:411–420. CrossRefGoogle Scholar
  9. 9.
    Agha-Rahimi A, Khalili MA, Nottola SA, Miglietta S, Moradi A (2016) Cryoprotectant-free vitrification of human spermatozoa in new artificial seminal fluid. Andrology 4:1037–1044. CrossRefGoogle Scholar
  10. 10.
    Chohan KR, Griffin JT, Lafromboise M, Jonge CJ, Carrell DT (2006) Comparison of chromatin assays for DNA fragmentation evaluation in human sperm. J Androl 27:53–59. CrossRefGoogle Scholar
  11. 11.
    Fernández JL, Muriel L, Goyanes V, Segrelles E, Gosálvez J, Enciso M, LaFromboise M, De Jonge C (2005) Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test. Fertil Steril 84:833–842. CrossRefGoogle Scholar
  12. 12.
    Ribas-Maynou J, García-Peiró A, Fernández-Encinas A, Abad C, Amengual MJ, Prada E, Navarro J, Benet J (2013) Comprehensive analysis of sperm DNA fragmentation by five different assays: TUNEL assay, SCSA, SCD test and alkaline and neutral Comet assay. Andrology 1:715–722. CrossRefGoogle Scholar
  13. 13.
    Fernández JL, Muriel L, Rivero MT, Goyanes V, Vazquez R, Alvarez JG (2003) The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation. J Androl 24:59–66. CrossRefGoogle Scholar
  14. 14.
    Cissen M, Wely MV, Scholten I, Mansell S, Bruin JP, Mol BW, Braat D, Repping S, Hamer G (2016) Measuring sperm DNA fragmentation and clinical outcomes of medically assisted reproduction: a systematic review and meta-analysis. PLoS ONE 11:e0165125. CrossRefGoogle Scholar
  15. 15.
    Kirkman-Brown JC, De Jonge C (2017) Sperm DNA fragmentation in miscarriage—a promising diagnostic, or a test too far? Reprod Biomed Online 34:3–4. CrossRefGoogle Scholar
  16. 16.
    Osman A, Alsomait H, Seshadri S, El-Toukhy T, Khalaf Y (2015) The effect of sperm DNA fragmentation on live birth rate after IVF or ICSI: a systematic review and meta-analysis. Reprod Biomed Online 30:120–127. CrossRefGoogle Scholar
  17. 17.
    Ioannou D, Tempest HG (2018) Does genome organization matter in spermatozoa? A refined hypothesis to awaken the silent vessel. Syst Biol Reprod Med 64:518–534. CrossRefGoogle Scholar
  18. 18.
    Practice Committee of the American Society for Reproductive Medicine (2013) The clinical utility of sperm DNA integrity testing: a guideline. Fertil Steril 99:673–677. CrossRefGoogle Scholar
  19. 19.
    Ménézo Y, Dale B, Cohen M (2010) DNA damage and repair in human oocytes and embryos: a review. Zygote 18:357–365. CrossRefGoogle Scholar
  20. 20.
    Vazharova R, Kremensky I (2016) Individual capacity for DNA repair and maintenance of genomic integrity: a fertile ground for studies in the field of assisted reproduction. Biotechnol Biotechnol Equip 30:419–433CrossRefGoogle Scholar
  21. 21.
    Marchetti F, Essers J, Kanaar R, Wyrobek AJ (2007) Disruption of maternal DNA repair increases sperm-derived chromosomal aberrations. Proc Natl Acad Sci USA 104:17725–17729. CrossRefGoogle Scholar
  22. 22.
    Gat I, Li N, Yasovich N, Antes R, Kuznyetsov V, Zohni K, Weizman NF, Librach C (2018) Sperm DNA fragmentation index does not correlate with blastocyst euploidy rate in egg donor cycles. Gynecol Endocrinol 34:212–216. CrossRefGoogle Scholar
  23. 23.
    Esbert M, Pacheco A, Vidal F, Florensa M, Riqueros M, Ballesteros A, Garrido N, Calderón G (2011) Impact of sperm DNA fragmentation on the outcome of IVF with own or donated oocytes. Reprod Biomed Online 23:704–710. CrossRefGoogle Scholar
  24. 24.
    Prapas N, Prapas Y, Panagiotidis Y, Prapa S, Vanderzwalmen P, Schoysman R, Makedos G (2005) GnRH agonist versus GnRH antagonist in oocyte donation cycles: a prospective randomized study. Hum Reprod 20:1516–1520. CrossRefGoogle Scholar
  25. 25.
    Prapas N, Tavaniotou A, Panagiotidis Y, Prapa S, Kasapi E, Goudakou M, Papatheodorou A, Prapas Y (2009) GnRH antagonists and endometrial receptivity in oocyte recipients: a prospective randomized trial. Reprod Biomed Online 18:276-281.
  26. 26.
    López G, Lafuente R, Checa MA, Carreras R, Brassesco M (2013) Diagnostic value of sperm DNA fragmentation and sperm high-magnification for predicting outcome of assisted reproduction treatment. Asian J Androl 15:790–794. CrossRefGoogle Scholar
  27. 27.
    Petousis S, Prapas Y, Papatheodorou A, Margioula-Siarkou C, Papatzikas G, Panagiotidis Y, Karkanaki A, Ravanos K, Prapas N (2018) Fluorescence in situ hybridisation sperm examination is significantly impaired in all categories of male infertility. Andrologia 50:e12847. CrossRefGoogle Scholar
  28. 28.
    Gorczyca W, Traganos F, Jesionowska H, Darzynkiewicz Z (1993) Presence of DNA strand breaks and increased sensitivity of DNA in situ to denaturation in abnormal human sperm cells: analogy to apoptosis of somatic cells. Exp Cell Res 207:202–205CrossRefGoogle Scholar
  29. 29.
    Rybar R, Kopecka V, Prinosilova P, Markova P, Rubes J (2011) Male obesity and age in relationship to semen parameters and sperm chromatin integrity. Andrologia 43:286–291. CrossRefGoogle Scholar
  30. 30.
    Tunc O, Bakos HW, Tremellen K (2011) Impact of body mass index on seminal oxidative stress. Andrologia 43:121–128. CrossRefGoogle Scholar
  31. 31.
    Chavarro JE, Toth TL, Wright DL, Meeker JD, Hauser R (2010) Body mass index in relation to semen quality, sperm DNA integrity, and serum reproductive hormone levels among men attending an infertility clinic. Fertil Steril 93:2222–2231. CrossRefGoogle Scholar
  32. 32.
    Fariello RM, Pariz JR, Spaine DM, Cedenho AP, Bertolla RP, Fraietta R (2012) Association between obesity and alteration of sperm DNA integrity and mitochondrial activity. B J U Int 110:863–867. CrossRefGoogle Scholar
  33. 33.
    Tandara M, Bajic A, Tandara L, Sunj M, Jurisic Z, Jukic M (2013) Correlation between proportions of sperm with DNA fragmentation assessed by Halosperm test and values of standard quality parameters of semen and possible impact on embryo quality. Zdrav Vestn 82:298–307Google Scholar
  34. 34.
    Vagnini L, Baruffi RLR, Mauri AL, Petersen CG, Massaro FC, Pontes A, Oliveira JB, Franco JG Jr (2007) The effects of male age on sperm DNA damage in an infertile population. Reprod Biomed Online 15:514–519CrossRefGoogle Scholar
  35. 35.
    Schmid TE, Eskenazi B, Baumgartner A, Marchetti F, Young S, Weldon R, Anderson D, Wyrobek AJ (2006) The effects of male age on sperm DNA damage in healthy non-smokers. Hum Reprod 22:180–187. CrossRefGoogle Scholar
  36. 36.
    Frattarelli JL, Miller KA, Miller BT, Elkind-Hirsch K, Scott RT (2008) Male age negatively impacts embryo development and reproductive outcome in donor oocyte assisted reproductive technology cycles. Fertil Steril 90:97–103. CrossRefGoogle Scholar
  37. 37.
    Kaarouch I, Bouamoud N, Madkour A, Louanjli N, Saadani B, Assou S, Aboulmaouahib S, Amzazi S, Copin H, Benkhalifa M, Sefrioui O (2018) Paternal age: negative impact on sperm genome decays and IVF outcomes after 40 years. Mol Reprod Dev. Google Scholar
  38. 38.
    Irvine DS, Twigg JP, Gordon EL, Fulton N, Milne PA, Aitken R (2000) DNA integrity in human spermatozoa: relationships with semen quality. J Androl 21:33–44Google Scholar
  39. 39.
    Sheikh N, Amiri I, Farimani M, Najafi R, Hadeie J (2008) Correlation between sperm parameters and sperm DNA fragmentation in fertile and infertile men. Int J Reprod Biomed (Yazd) 6:13–18Google Scholar
  40. 40.
    Avendaño C, Oehninger S (2011) DNA fragmentation in morphologically normal spermatozoa: how much should we be concerned in the ICSI era? J Androl 32:356–363. CrossRefGoogle Scholar
  41. 41.
    Belloc S, Benkhalifa M, Cohen-Bacrie M, Dalleac A, Chahine H, Amar E, Zini A (2014) Which isolated sperm abnormality is most related to sperm DNA damage in men presenting for infertility evaluation. J Assist Reprod Genet 31:527–532. CrossRefGoogle Scholar
  42. 42.
    Rafighdoost H, Farsi MM, Javadi M, Khafri S (2013) Relationship between sperm parameters and DNA fragmentation using a halosperm kit. Anat Sci J 10:79–86Google Scholar
  43. 43.
    Enciso M, Iglesias M, Galán I, Sarasa J, Gosálvez A, Gosálvez J (2011) The ability of sperm selection techniques to remove single-or double-strand DNA damage. Asian J Androl 13:764–768. CrossRefGoogle Scholar
  44. 44.
    Zini A, Sigman M (2009) Are tests of sperm DNA damage clinically useful? Pros and cons. J Androl 30:219–229. CrossRefGoogle Scholar
  45. 45.
    Yılmaz S, Zergeroğlu AD, Yılmaz E, Sofuoglu K, Delikara N, Kutlu P (2010) Effects of sperm DNA fragmentation on semen parameters and ICSI outcome determined by an improved SCD test, Halosperm. Int J Fertil Steril 4:73–78Google Scholar
  46. 46.
    Larson-Cook KL, Brannian JD, Hansen KA, Kasperson KM, Aamold ET, Evenson DP (2003) Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay. Fertil Steril 80:895–902. CrossRefGoogle Scholar
  47. 47.
    Morris ID, Ilott S, Dixon L, Brison DR (2002) The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (Comet assay) and its relationship to fertilization and embryo development. Hum Reprod 17:990–998CrossRefGoogle Scholar
  48. 48.
    Høst E, Lindenberg S, Smidt-Jensen S (2000) The role of DNA strand breaks in human spermatozoa used for IVF and ICSI. Acta Obstet Gynecol Scand 79:559–563. CrossRefGoogle Scholar
  49. 49.
    Muriel L, Garrido N, Fernández JL, Remohí J, Pellicer A, de los Santos MJ, Meseguer M (2006) Value of the sperm deoxyribonucleic acid fragmentation level, as measured by the sperm chromatin dispersion test, in the outcome of in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril 85:371–383. CrossRefGoogle Scholar
  50. 50.
    Zhylkova I, Feskov O, Feskova I, Fedota O, Feskov V (2014) Sperm DNA fragmentation as a factor of male low reproductive function in IVF practice. Int J Biol 6:75–81Google Scholar
  51. 51.
    Twigg JP, Irvine DS, Aitken RJ (1998) Oxidative damage to DNA in human spermatozoa does not preclude pronucleus formation at intracytoplasmic sperm injection. Hum Reprod 13:1864–1871CrossRefGoogle Scholar
  52. 52.
    Simon L, Murphy K, Shamsi MB, Liu L, Emery B, Aston KI, Hotaling J, Carrell DT (2014) Paternal influence of sperm DNA integrity on early embryonic development. Hum Reprod 29:2402–2412. CrossRefGoogle Scholar
  53. 53.
    Virro MR, Larson-Cook KL, Evenson DP (2004) Sperm chromatin structure assay (SCSA®) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril 81:1289–1295CrossRefGoogle Scholar
  54. 54.
    Fatehi AN, Bevers MM, Schoevers E, Roelen BAJ, Colenbrander B, Gadella BM (2006) DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after the first cleavages. J Androl 27:176–188. CrossRefGoogle Scholar
  55. 55.
    Sakkas D, Urner F, Bianchi PG, Bizzaro D, Wagner I, Jaquenoud N, Manicardi G, Campana A (1996) Sperm chromatin anomalies can influence decondensation after intracytoplasmic sperm injection. Hum Reprod 11:837–843CrossRefGoogle Scholar
  56. 56.
    Bungum M, Humaidan P, Spano M, Jepson K, Bungum L, Giwercman A (2004) The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI. Hum Reprod 19:1401–1408. CrossRefGoogle Scholar
  57. 57.
    Henkel R, Hajimohammad M, Stalf T, Hoogendijk C, Mehnert C, Menkveld R, Gips H, Schill WB, Kruger TF (2004) Influence of deoxyribonucleic acid damage on fertilization and pregnancy. Fertil Steril 81:965–972. CrossRefGoogle Scholar
  58. 58.
    Gandini L, Lombardo F, Paoli D, Caruso F, Eleuteri P, Leter G, Ciriminna R, Culasso F, Dondero F, Lenzi A, Spanò M (2004) Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage. Hum Reprod 19:1409–1417. CrossRefGoogle Scholar
  59. 59.
    Borini A, Tarozzi N, Nadalini M (2017) Sperm DNA fragmentation testing in male infertility work-up: are we ready? Transl Androl Urol 6:S580–S582 CrossRefGoogle Scholar
  60. 60.
    Simon L, Lutton D, McManus J, Lewis SE (2011) Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success. Fertil Steril 95:652–657. CrossRefGoogle Scholar
  61. 61.
    Muratori M, Tarozzi N, Cambi M, Boni L, Iorio AL, Passaro C, Luppino B, Nadalini M, Marchiani S, Tamburrino L, Forti G, Maggi M, Baldi E, Borini A (2016) Variation of DNA fragmentation levels during density gradient sperm selection for assisted reproduction techniques: a possible new male predictive parameter of pregnancy? Medicine (Baltimore) 95:e3624. CrossRefGoogle Scholar
  62. 62.
    Jayaraman V, Upadhya D, Narayan PK, Adiga SK (2012) Sperm processing by swim-up and density gradient is effective in elimination of sperm with DNA damage. J Assist Reprod Genet 29:557–563. CrossRefGoogle Scholar
  63. 63.
    Malvezzi H, Sharma R, Agarwal A, Abuzenadah AM, Abu-Elmagd M (2014) Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial. Reprod Biol Endocrinol 12:121. CrossRefGoogle Scholar
  64. 64.
    Shafik A, Shafik AA, Shafik I, El Sibai O (2006) Molecular andrology as related to sperm DNA fragmentation/sperm chromatin biotechnology. Arch Androl 52:299–310. CrossRefGoogle Scholar
  65. 65.
    Lin MH, Kuo-Kuang Lee R, Li SH, Lu CH, Sun FJ, Hwu YM (2008) Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril 90:352–359. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Sevastiani Antonouli
    • 1
    Email author
  • Achilleas Papatheodorou
    • 2
  • Yannis Panagiotidis
    • 2
  • Stamatios Petousis
    • 2
  • Nikos Prapas
    • 2
  • Stefania Annarita Nottola
    • 3
  • Maria Grazia Palmerini
    • 1
  • Guido Macchiarelli
    • 1
  • Yannis Prapas
    • 2
  1. 1.Department of Life, Health and Environmental SciencesUniversity of L’AquilaL’AquilaItaly
  2. 2.Iakentro Fertility Center, IVF LaboratoryThessalonikiGreece
  3. 3.Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, Laboratory of Electron Microscopy “Pietro M. Motta”University La SapienzaRomeItaly

Personalised recommendations