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Cited by 56Sperm and Testicular FunctionMale Reproductive Health StudiesReproductive Biology and Fertility

Teratozoospermia: Its association with sperm DNA defects, apoptotic alterations, and oxidative stress

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Ammar, Oumaima, Mehdi, Meriem, Muratori, Monica·2020·Andrology
DOI 10.1111/andr.12778
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Abstract

This study was carried out to evaluate the level of nuclear sperm DNA damage in men with isolated polymorphic teratozoospermia and examining its relationship with apoptosis and oxidative stress. A total of 89 subjects were divided into two groups: men with isolated teratozoospermia (n = 69) and men with normal semen parameters (n = 20) as controls. Sperm DNA breaks were determined by using acridine orange staining. The proportion of viable spermatozoa with mitochondrial transmembrane depolarization was detected by fluorescence microscopy through the use of MitoPTJC‐1 staining method. Bivariate Annexin V/6‐CFDA analysis was then set out in order to measure the percentage of both viable and dead spermatozoa with phosphatidylserine (PS) externalization. Seminal antioxidant profile (reduced glutathione (GSHr); oxidized glutathione (GSSG); glutathione S‐transferase (GST)) and total protein sulfhydryl (P‐SH) concentrations were measured spectrophotometrically. Men with isolated teratozoospermia, when compared to controls, showed significantly increased level of single sperm DNA breaks and higher proportions of spermatozoa with phosphatidylserine externalization and mitochondrial depolarization. Among the differently studied oxidative stress seminal parameters, the rates of seminal GSHr, GST, and P‐SH were significantly decreased in the teratozoospermic group. However, the seminal rates of GSSG and GST have decreased, but only GST did not show a significant difference. Interestingly, significant correlations were found between the studied apoptotic markers and the rate of atypical sperm forms with the incidences of head abnormalities. Furthermore, positive inter‐correlations were found between sperm DNA defects, impaired seminal antioxidant status, and the apoptotic sperm markers. Such data provide clear evidence that the apoptotic alterations are closely correlated to abnormal sperm morphology and DNA damage. Moreover, decreased seminal antioxidant profile may be a crucial factor involved in the mechanism of sperm cell death‐mediated DNA breaks in teratozoospermic semen.

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  • Pathophysiological studies have shown sometimes controversial associations between sperm morphology and sperm competence (aptitude to undergo the acrosomal reaction, DNA integrity, chromatin condensation, etc.).

    “Sperm morphology assessment has also been examined in numerous pathophysiological studies that have shown sometimes controversial associations between morphology and sperm competence (aptitude to undergo the acrosomal reaction, DNA integrity, chromatin condensation, etc.) [5, 6, 7, 8].”

    From Recommendations for Human Sperm Morphology Assessment in 2025: An Expert Review From the French BLEFCO Group (2026) · cites this paper as [8]

    The Discussion (page unavailable) reports an association between abnormal sperm morphology and DNA damage, while also noting studies that found no significant association. The Introduction links teratozoospermia with chromatin decondensation. Our copy does not establish an association between morphology and the ability to undergo the acrosomal reaction. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

  • Some studies have shown a correlation between sperm morphological defects and nuclear genetic defects, apoptotic alterations, and reactive oxidative stress, which may negatively impact fertility potential.

    “Some studies have shown a correlation between sperm morphological defects and nuclear genetic defects, apoptotic alterations, and reactive oxidative stress, which may negatively impact fertility potential (13-15).”

    From Isolated teratozoospermia: revisiting its relevance in male infertility: a narrative review (2024) · cites this paper as [14]

  • Abnormal sperm morphology in general has shown a correlation with changes in genetic materials, increased rate of apoptosis, overproduction of oxidative stress, and a decrease in antioxidant activities.

    “While some morphological defects are associated with specific functional deficiencies, such as those with tail defects that result in abnormal motility, abnormal sperm morphology, in general, has shown a correlation with changes in genetic materials, increased rate of apoptosis, overproduction of oxidative stress as well as a decrease in antioxidant activities (13-15).”

    From Isolated teratozoospermia: revisiting its relevance in male infertility: a narrative review (2024) · cites this paper as [14]

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Research works printed but not linked: 75

  1. Brahem S, Mehdi M, Elghezal H, Saad A. Detection of DNA frag- mentation and meiotic segregation in human with isolated terato- zoospermia. J Assist Reprod Genet. 2011;28:41-48.
  2. Mehdi M, Gmidène A, Brahem S, Guerin JF, Elghezal H, Saad A. Aneuploidy rate in spermatozoa of selected men with severe tera- tozoospermia. Andrologia. 2012;1:139-143.
  3. Ammar O, Tesnim A, Zohra H, et al. Investigation on the origin of sperm morphological defects: oxidative attacks, chromatin imma- turity, and DNA fragmentation. Environ Sci Pollut Res. 2018;25:1-12.
  4. Ammar O, Haouas Z, Hamouda B, et al. Relationship between sperm DNA damage with sperm parameters, oxidative markers in terato- zoospermic men. Eur J Obstet Gynecol Reprod Biol. 2018;233:70-75.
  5. WHO. World Health Organization Laboratory Manual for the Examination and Processing of Human. Semen. WHO Press; 2010.
  6. Kruger TF, Menkveld R, Stander FSH, et al. Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertil Steril. 1986;46:1118-1123.
  7. Auger J, Eustash F. Standardisation de la classification mor- phologique des spermatozoïdes humains selon la méthode de David modifiée. Andrologie. 2000;10(4):358-373.
  8. Ajina T, Ammar O, Haouas Z, et al. Assessment of human sperm DNA integrity using two cytochemical tests: Acridine orange test and toluidine blue assay. Andrologia. 2017;49(10):e12765.
  9. Atig F, Kerkeni A, Saad A, Ajina M. Effects of reduced semi- nal enzymatic antioxidants on sperm DNA fragmentation and semen quality of Tunisian infertile men. J Assist Reprod Genet. 2017;34:373-381.
  10. Hichri R, Amor H, Khammari M, et al. Apoptotic sperm biomarkers and the correlation between conventional sperm parameters and clinical characteristics. Andrologia. 2018;50:1-9.
  11. Hammadeh ME, Zavos PM, Rosenbaum P, Schmidt W. Comparison between the quality and function of sperm after semen processing with two different methods. Asian J Androl. 2001;3(2):125-130.
  12. Mortimer D, Menkveld R. Sperm morphology assessment -his- torical perspectives and current opinions andrology lab corner. J Androl. 2001;22(2):192-205.
  13. Lewis SE, John Aitken R, Conner SJ, et al. The impact of sperm DNA damage in assisted conception and beyond: Recent advances in di- agnosis and treatment. Reprod Biomed Online. 2013;27(4):325-337.
  14. Al-Dujaily SS, Al-Jnabi MH, Jasim SN. The influence of Leukocytospermia and Teratozoospermia in IUI outcome. Int J Adv Res. 2015;3:255-265.
  15. Sakkas D, Mariethoz E, Manicardi G, Bizzaro D, Bianchi PG, Bianchi U. Origin of DNA damage in ejaculated human spermatozoa. Rev Reprod. 1999;4(1):31-37.
  16. Aitken RJ, De Iuliis GN. Origins and consequences of DNA damage in male germ cells. Reprod Biomed Online. 2007;14(6):727-733.
  17. Aitken RJ, Bronson R, Smith TB, De Iuliis GN. The source and significance of DNA damage in human spermatozoa; a commen- tary on diagnostic strategies and straw man fallacies. Mol Hum. 2013;19(8):475-485.
  18. Aitken RJ, West KM. Analysis of the relationship between reac- tive oxygen species production and leucocyte infiltration in frac- tions of human semen separated on Percoll gradients. Int J Androl. 1990;13(6):433-451.
  19. Aitken RJ, Baker MA, Sawyer D. Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic dis- ease. Reprod Biomed Online. 2003;7(1):65-70.
  20. Said TM, Agarwal A, Sharma RK, Thomas AJ Jr, Sikka SC. Impact of sperm morphology on DNA damage caused by oxidative stress induced by β-nicotinamide adenine dinucleotide phosphate. Fertil Steril. 2005;83(1):95-103.
  21. Said TM, Grunewald S, Paasch U, et al. Advantage of combin- ing magnetic cell separation with sperm preparation techniques. Reprod Biomed Online. 2005;10(6):740-746.
  22. Ammar O, Mehdi M, Tekeya O, Neffati F, Haouas Z. Novel associ- ation between apoptotic sperm biomarkers with seminal biochem- ical parameters and acetylcholinesterase activity in patients with teratozoospermia. J Assist Reprod Genet. 2019;36:2367-2378.
  23. Aziz N, Said T, Paasch U, Agarwal A. The relationship between human sperm apoptosis, morphology and the sperm deformity index. Hum Reprod. 2007;22(5):1413-1419.
  24. Marchiani S, Tamburrino L, Olivito B, et al. Characterization and sorting of flow cytometric populations in human semen. Andrology. 2014;2:394-401.
  25. Paasch U, Agarwal A, Gupta AK, et al. Apoptosis signal transduction and the maturity status of human spermatozoa. Ann N Y Acad Sci. 2003;1010(1):486-488.
  26. Brugnon F, Ouchchane L, Verheyen G, et al. Fluorescence micros- copy and flow cytometry in measuring activated caspases in human spermatozoa. Int J Androl. 2009;32:265-273.
  27. David G, Bisson JP, Czyglick F, Jouannet P, Gernigon C. Anomalies morphologiques du spermatozoïde humain. 1) Propositions pour un système de classification. J Gynecol Obstet Biol Reprod. 1975;1:17-36.
  28. Kemal Duru N, Morshedi M, Oehninger S. Effects of hydrogen per- oxide on DNA and plasma membrane integrity of human spermato- zoa. Fertil Steril. 2000;74(6):1200-1207.
  29. Tejada RI, Mitchell JC, Norman A, Marik JJ, Friedman S. A test for the practical evaluation of male fertility by acridine orange (AO) flu- orescence. Fertil Steril. 1984;42(1):87-91.
  30. Srivastava SK, Beutler E. Accurate measurement of oxidized gluta- thione content of human, rabbit, and rat red blood cells and tissues. Anal Biochem. 1968;25(1):70-76.
  31. Akerboom TPM, Sies H. Assay of glutathione, glutathione dis- sulfide, and glutathione mixed dissulfides in biological samples. Methods Enzymol. 1981;77:373.
  32. Habig WH, Jakoby WB. Assays for Differentiation of Glutathione S-Transferases. Methods Enzymol. 1981;77:398-405.
  33. Kasperczyk A, Dobrakowski M, Czuba ZP, Kapka-Skrzypczak L, Kasperczyk S. Environmental exposure to zinc and copper in- fluences sperm quality in fertile males. Ann Agric Environ Med. 2016;23:138-143.
  34. Braham A, Ghedir H, Zidi I, et al. Nuclear sperm quality in total polymorphic teratozoospermia and its impact on intracytoplasmic sperm injection outcome. Andrologia. 2019;51:1-10.
  35. Marcon L, Boissonneault G. Transient DNA strand breaks during mouse and human spermiogenesis: new insights in stage specificity and link to chromatin remodeling. Biol Reprod. 2004;70(4):910-918.
  36. Aitken RJ, Smith TB, Jobling MS, Baker MA, De Iuliis GN. Oxidative stress and male reproductive health. Asian J Androl. 2014;16:31-38.
  37. Muratori M, Tamburrino L, Marchiani S, Kar S, Samanta L. Investigation on the origin of sperm DNA fragmentation: role of apoptosis, immatu- rity and oxidative stress. Mol Med. 2015;21:109-122.
  38. Perrin A, Louanjli N, Ziane Y, et al. Study of aneuploidy and DNA fragmentation in gametes of patients with severe teratozoosper- mia. Reprod Biomed Online. 2011;22:148-154.
  39. Skowronek F, Casanova G, Alciaturi J, et al. DNA sperm damage correlates with nuclear ultrastructural sperm defects interatozo- ospermic men. Andrologia. 2012;44(1):59-65.
  40. Cho C, Willis WD, Goulding EH, et al. Haploinsufficiency of prota- mine-1 or -2 causes infertility in mice. Nat Genet. 2001;28:82-86.
  41. Zini A, Phillips S, Courchesne A, et al. Sperm head morphology is related to high deoxyribonucleic acid stainability assessed by sperm chromatin structure assay. Fertil Steril. 2009;91:2495-2500.
  42. Chan PJ, Corselli JU, Patton WC, &, . A simple comet assay for archived sperm correlates DNA fragmentation to reduced hyperactivation and penetration of zona-free hamster oocytes. Fertil Steril. 2001;75:186-192.
  43. Donnelly ET, McClure N, Lewis SE. Assessment ofDNA integ- rity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation. Hum Reprod. 2001;16:1191-1199.
  44. Barroso G, Morshedi M, Oehninger S. Analysis of DNA frag- mentation, plasma membrane translocation of phosphatidylser- ine and oxidative stress in human spermatozoa. Hum Reprod. 2000;15(6):1338-1344.
  45. Oosterhuis GJE, Mulder AB, Kalsbeek-Batenburg E, Beschmann H. Measuring apoptosis in human spermatozoa: A biological assay for semen quality? Fertil Steril. 2000;13(2):353-359.
  46. Shen H-M, Dai J, Chia S-E, Lim A, Ong CN. Detection of apoptotic alterations in sperm in subfertile patients and their correlations with sperm quality. Hum Reprod. 2002;17(5):1266-1273.
  47. Barroso G, Taylor S, Morshedi M, Manzur F, Gaviño F, Oehninger S. Mitochondrial membrane potential integrity and plasma membrane trans- location of phosphatidylserine as early apoptotic markers: A comparison of two different sperm subpopulations. Fertil Steril. 2006;85(1):149-154.
  48. Koppers AJ, De Iuliis GN, Finnie JM, McLaughlin EA, Aitken RJ. Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa. J Clin Endocrinol Metab. 2008;93(8):3199-3207.
  49. Zhang HB, Lu SM, Wang MCY, L, Li X & Chen ZJ, . Early apoptotic changes in human spermatozoa and their relationships with con- ventional semen parameters and sperm DNA fragmentation. Asian J Androl. 2008;10(2):227-235.
  50. Mignotte B, Vayssiere JL. Mitochondrial control of apoptosis. Adv Cell Aging Gerontol. 2001;5:93-122.
  51. Suen DF, Norris KL, Youle RJ. Mitochondrial dynamics and apopto- sis. Genes Dev. 2008;22(12):1577-1590.
  52. Ochsendorf FR, Buhl R, Bästlein A, Beschmann H. Glutathione in spermatozoa and seminal plasma of infertile men. Hum Reprod. 1998;13:353-359.
  53. Chaudhari AR, Piyali D, Ramji S. Study of oxidative stress and re- duced glutathione levels in seminal plasma of human subjects with different fertility potential. Biomed Res. 2008;3:207-210.
  54. Nabil H, Moemen LA, Abu Elela MH. Studying the levels of malond- ialdehyde and antioxidant parameters innormal and abnormal human seminal plasma. Aust J Basic Appl Sci. 2008;2(3):773-778.
  55. Atig F, Raffa M, Ali HB, Abdelhamid K, Saad A, Ajina M. Altered an- tioxidant status and increased lipid per-oxidation in seminal plasma of Tunisian infertile men. Int J Biol Sci. 2011;8:139-149.
  56. Atig F, Raffa M, Ali HB, Kerkeni A, Saad A, Ajina M. Impact of seminal trace element and glutathione levels on semen quality of Tunisian infertile men. BMC Urol. 2012;12:2-9.
  57. Al-azzawie HF, Naeim M, Saleman ED. Evaluation of enzymatic and non-enzymatic antioxidant status in seminal plasma of Iraqi Infertile Men. Int J Adv Res. 2014;2:158-167.
  58. Ebisch IM, Thomas CM, Peters WH, Braat DD, Steegers-Theunissen RP. The importance of folate, zinc and antioxidants in the pathogenesis and prevention of subfertility. Hum Reprod Update. 2007;13(2):163-174.
  59. Raijmakers MTM, Roelofs HMJ, Steegers EAP, et al. Glutathione and glutathione S-transferases A1-1 and P1-1 in seminal plasma may play a role in protecting against oxidative damage to spermato- zoa. Fertil Steril. 2003;79(1):169-172.
  60. Lenzi A, Picardo M, Gandini L, et al. Glutathione treatment of dyspermia: Effect on the lipoperoxidation process. Hum Reprod. 1994;9(11):2044-2050.
  61. Sorensen MB, Stoltenberg M, Danscher G, Ernst E. Chelation of intracellular zinc ions affects human sperm cell motility. Mol Hum Reprod. 1999;5(4):338-341.
  62. Chen SS, Chang LS, Wei YH. Oxidative damage to proteins and de- crease of antioxidant capacity in patients with varicocele. Free Radic Biol Med. 2001;30(11):1328-1334.
  63. Smith CV, Jones DP, Guenthner TM, Lash LH, Lauterburg BH. Compartmentation of glutathione: Implications for the study of toxicity and disease. Toxicol Appl Pharmacol. 1996;140(1):1-12.
  64. Mohanty G, Swain N, Goswami C. Histone retention, protein carbo- nylation, and lipid peroxidation in spermatozoa: Possible role in re- current pregnancy loss. Syst Biol Reprod Med. 2016;62(3):201-212.
  65. Kasperczyk A, Dobrakowski M, Czuba ZP, Horak S, Kasperczyk S. Environmental exposure to lead induces oxidative stress and modulates the function of the antioxidant defense system and the immune system in the semen of males with normal semen profile. Toxicol Appl Pharmacol. 2015;284(3):339-344.
  66. Kasperczyk A, Dobrakowski M, Horak S, Zalejska-Fiolka & J Birkner E. The influence of macro and trace elements on sperm quality. J Trace Elem Med Biol. 2015;30:153-159.
  67. Greco E, Romano S, Iacobelli M, et al. ICSI in cases of sperm DNA damage: Beneficial effect of oral antioxidant treatment. Hum Reprod. 2005;20(9):2590-2594.
  68. Showell M, Brown J, Yazdani A, Stankiewicz T, Hart J. Antioxidants for male subfertility (Review). Cochrane Libr. 2012. https://doi. org/10.1002/14651 858.CD007 411.pub3. · DOI · Paper page
  69. Talevi R, Barbato V, Fiorentino I, I, Braun S, Longobardi S, Gualtieri R, . Protective effects of in vitro treatment with zinc, d-aspartate and coenzyme q10 on human sperm motility, lipid peroxidation and DNA fragmentation. Reprod Biol Endocrinol. 2013;11:81.
  70. Ammar O, Houas Z, Mehdi M. The association between iron, cal- cium, and oxidative stress in seminal plasma and sperm. Environ Sci Pollut Res. 2019;26(14):14097-14105.
  71. Said TM, Paasch U, Glander HJ, Agarwal A. Role of caspases in male infertility. Hum Reprod Update. 2004;10(1):39-51.
  72. Agarwal A, Said TM. Oxidative stress, DNA damage and apoptosis in male infertility: A clinical approach. BJU Int. 2005;95:503-507.
  73. Fadi B, Choucaira E, Rachkidia G, et al. High level of DNA fragmen- tation in sperm of Lebanese infertile men using Sperm Chromatin Dispersion test. Middle East Fertil Soc J. 2016;21:269-276.
  74. Fatemeh D, Nasim T, Farzaneh F, Fatemeh Y, Ali Reza T. Evaluation of sperm protamine deficiency and apoptosis in infertile men with idiopathic teratozoospermia. Clin Exp Reprod Med. 2017;44:2.
  75. Zini A, San Gabriel M, Baazeem A. Antioxidants and sperm DNA damage: A clinical perspective. J Assist Reprod Genet. 2009;26(8):427-432.

Cited by

Sorted by citations

Authors
Ammar, O., Mehdi, M., & Muratori, M.
Affiliations
  • Laboratory of Histology Embryology and Cytogenetics (LR 18 ES 40) Faculty of Medicine University of Monastir Monastir Tunisia
  • University of Monastir (TN)
  • University of Florence (IT)
ORCID
Venue
Andrology
Publisher
Wiley
Published
2020-03-12
Volume
8
Issue
5
Pages
1095-1106
ISSN
  • 2047-2919 (ISSN-L)
  • 2047-2927
DOI
10.1111/andr.12778
OpenAlex
W3007702120
Cited by
56
References
3
Citations by year
2026: 9 · 2025: 14 · 2024: 5 · 2023: 13 · 2022: 9 · 2021: 4 · 2020: 2
FWCI
6.17
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • Andrology (publishedVersion)
  • PubMed (publishedVersion)
  • Florence Research (University of Florence) (submittedVersion, cc-by-nd)
Funders
Ministry of Higher Education and Scientific Research
Topics
  • Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
  • Health Sciences › Health Professions › General Health Professions › Male Reproductive Health Studies
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Reproductive Biology and Fertility
Keywords
teratozoospermia, sperm DNA damage, sperm apoptosis, oxidative stress, glutathione, glutathione S-transferase, mitochondrial depolarization, phosphatidylserine exposure, sperm morphology, male infertility
MeSH terms
Teratozoospermia, Adult, Antioxidants, Humans, Male, Mitochondria, Nucleic Acid Denaturation, Phosphatidylserines, Prospective Studies, Spermatozoa, Sulfhydryl Compounds, Biomarkers, Apoptosis, Oxidative Stress, DNA Breaks, Semen Analysis

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