Role of Sperm–Hyaluronic Acid Binding in the Evaluation and Treatment of Subfertile Men with ROS-Affected Semen: Assessment of Sperm with Oxidative Damage and HA-Mediated ICSI Sperm Selection

  • Ciler Celik-OzenciEmail author
  • Gabor Huszar


Excessive levels of free radicals diminish the functional integrity of spermatozoa. Levels of reactive oxygen species (ROS) produced by spermatozoa were negatively correlated with the quality of sperm in the original semen. “Intrinsic,” “extrinsic,” and “iatrogenic” sources of ROS production were identified within semen: Intrinsic sources of ROS in semen are morphologically abnormal and arrested maturity spermatozoa and leukocytes. Poor sperm quality showing attributes of arrested sperm maturation is linked to increased ROS generation as a consequence of excess residual cytoplasm related to the arrest of cytoplasmic extrusion in terminal spermiogenesis. In the past 20 years, the Huszar lab has studied several key events of sperm maturation, including cytoplasmic extrusion and expression of the HspA2 chaperone protein. In experiments related to sperm function and fertilizing potential, Huszar et al. have established that, simultaneously with cytoplasmic extrusion during terminal spermiogenesis, there is a remodeling of the plasma membrane that facilitates the formation of the zona pellucida- and hyaluronic acid (HA)-binding sites. The studies with HA immobilized to glass slides or Petri dishes showed that sperm firmly bind to HA. However, not all sperm exhibited HA binding ability. These data supported the hypothesis that the ability of sperm–HA binding is related to sperm cellular maturity.


Sperm–hyaluronic acid binding Sperm function Oxidative stress Reactive oxygen ­species Male infertility Intracytoplasmic sperm selection DNA chain degradation 


  1. 1.
    Aitken RJ, Koopman P, Lewis SE. Seeds of concern. Nature. 2004;432:48–52.PubMedGoogle Scholar
  2. 2.
    De Laminardi E, Tsai C, Harakat A, et al. Involvement of reactive oxygen species in human sperm acrosome reaction induced by A 23187, lysophosphatidylcholine, and biological Xuid ultraWltrates. J Androl. 1998;19:585–94.Google Scholar
  3. 3.
    de Lamirande E, Gagnon C. Human sperm hyperactivation and capacitation as parts of an oxidative process. Free Radic Biol Med. 1993;14:157–66.PubMedGoogle Scholar
  4. 4.
    Aitken RJ, Paterson M, Fisher H, et al. Redox regulation of tyrosine phosphorylation in human spermatozoa and its role in the control of human sperm function. J Cell Sci. 1995;108:2017–25.PubMedGoogle Scholar
  5. 5.
    Aitken RJ, Gordon E, Harkiss D, et al. Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa. Biol Reprod. 1998;59:1037–46.PubMedGoogle Scholar
  6. 6.
    Gomez E, Irvine DS, Aitken RJ. Evaluation of a spectrophotometric assay for the measurement of malondialdehyde and 4-hydroxyalkenals in human spermatozoa: relationships with semen quality and sperm function. Int J Androl. 1998;21:81–94.PubMedGoogle Scholar
  7. 7.
    Aitken J, Krausz C, Buckingham D. Relationships between biochemical markers for residual sperm cytoplasm, reactive oxygen species generation, and the presence of leukocytes and precursor germ cells in human sperm suspensions. Mol Reprod Dev. 1994;39:268–79.PubMedGoogle Scholar
  8. 8.
    Huszar G, Vigue L. Correlation between the rate of lipid peroxidation and cellular maturity as measured by creatine kinase activity in human spermatozoa. J Androl. 1994;15:71–7.PubMedGoogle Scholar
  9. 9.
    Huszar G, Corrales M, Vigue L. Correlation between sperm creatine phosphokinase activity and sperm concentrations in normospermic and oligospermic men. Gamete Res. 1988;19:67–75.PubMedGoogle Scholar
  10. 10.
    Huszar G, Vigue L, Corrales M. Sperm creatine phosphokinase activity as a measure of sperm quality in normospermic, variablespermic, and oligospermic men. Biol Reprod. 1988;38:1061–6.PubMedGoogle Scholar
  11. 11.
    Huszar G, Vigue L. Spermatogenesis-related change in the synthesis of the creatine kinase B-type and M-type isoforms in human spermatozoa. Mol Reprod Dev. 1990;25:258–62.PubMedGoogle Scholar
  12. 12.
    Aziz N, Saleh RA, Sharma RK. et al; Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index. Fertil Steril. 2004;81:349–54.PubMedGoogle Scholar
  13. 13.
    Sikka SC. Relative impact of oxidative stress on male reproductive function. Curr Med Chem (Rev). 2001;8:851–62.Google Scholar
  14. 14.
    Huszar G, Vigue L. Incomplete development of human spermatozoa is associated with increased creatine phosphokinase concentration and abnormal head morphology. Mol Reprod Dev. 1993;34:292–8.PubMedGoogle Scholar
  15. 15.
    Huszar G, Stone K, Dix D, et al. Putative creatine kinase M-isoform in human sperm is identified as the 70-kilodalton heat shock protein HspA2. Biol Reprod. 2000;63:925–32.PubMedGoogle Scholar
  16. 16.
    Lalwani S, Sayme N, Vigue L, et al. Biochemical markers of early and late spermatogenesis: relationship between the lactate dehydrogenase-X and creatine kinase-M isoform concentrations in human spermatozoa. Mol Reprod Dev. 1996;43:495–502.PubMedGoogle Scholar
  17. 17.
    Huszar G, Vigue L, Corrales M. Sperm creatine kinase activity in fertile and infertile oligospermic men. J Androl. 1990;11:40–6.PubMedGoogle Scholar
  18. 18.
    Huszar G, Vigue L, Morshedi M. Sperm creatine phosphokinase M-isoform ratios and fertilizing potential of men: a blinded study of 84 couples treated with in vitro fertilization. Fertil Steril. 1992;57:882–8.PubMedGoogle Scholar
  19. 19.
    Huszar G, Vigue L, Oehninger S. Creatine kinase immunocytochemistry of human sperm-hemizona complexes: selective binding of sperm with mature creatine kinase-staining pattern. Fertil Steril. 1994;61:136–42.PubMedGoogle Scholar
  20. 20.
    Huszar G, Sbracia M, Vigue L, et al. Sperm plasma membrane remodeling during spermiogenetic maturation in men: relationship among plasma membrane beta 1,4-galactosyltransferase, cytoplasmic creatine phosphokinase, and creatine phosphokinase isoform ratios. Biol Reprod. 1997;56:1020–4.PubMedGoogle Scholar
  21. 21.
    Huszar G, Willetts M, Corrales M. Hyaluronic acid (Sperm Select) improves retention of sperm motility and velocity in normospermic and oligospermic specimens. Fertil Steril. 1990;54:1127–34.PubMedGoogle Scholar
  22. 22.
    Sbracia M, Grasso J, Sayme N, et al. Hyaluronic acid substantially increases the retention of motility in cryopreserved/thawed human spermatozoa. Hum Reprod. 1997;12:1949–54.PubMedGoogle Scholar
  23. 23.
    Huszar G, Ozenci CC, Cayli S, et al. Hyaluronic acid binding by human sperm indicates cellular maturity, viability, and unreacted acrosomal status. Fertil Steril. 2003;79:1616–24.PubMedGoogle Scholar
  24. 24.
    Huszar G, Jakab A, Sakkas D, et al. Fertility testing and ICSI sperm selection by hyaluronic acid binding: clinical and genetic aspects. RBM Online. 2007;14:650–63.PubMedGoogle Scholar
  25. 25.
    Sharma RK, Pasqualotto AE, Nelson DR, et al. Relationship between seminal white blood cell counts and oxidative stress in men treated at an infertility clinic. J Androl. 2001;22:575–83.PubMedGoogle Scholar
  26. 26.
    Iwasaki A, Gagnon C. Formation of reactive oxygen species in spermatozoa of infertile patients. Fertil Steril. 1992;57:409–16.PubMedGoogle Scholar
  27. 27.
    Shekarriz M, Thomas Jr AJ, Agarwal A. Incidence and level of seminal reactive oxygen species in normal men. Urology. 1995;45:103–7.PubMedGoogle Scholar
  28. 28.
    Shekarriz M, DeWire DM, Thomas Jr AJ, et al. A method of human semen centrifugation to minimize the iatrogenic sperm injuries caused by reactive oxygen species. Eur Urol. 1995;28:31–5.PubMedGoogle Scholar
  29. 29.
    Potts JM, Sharma R, Pasqualotto F, et al. Association of ureaplasma urealyticum with abnormal reactive oxygen species levels and absence of leukocytospermia. J Urol. 2000;163:1775–8.PubMedGoogle Scholar
  30. 30.
    Potts RJ, Notarianni LJ, Jefferies TM. Seminal plasma reduces exogenous oxidative damage to human sperm, determined by the measurement of DNA strand breaks and lipid peroxidation. Mutat Res. 2000;447:249–56.PubMedGoogle Scholar
  31. 31.
    Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci. 2000;60–61:481–92.PubMedGoogle Scholar
  32. 32.
    Irvine DS, Twigg JP, Gordon EL, et al. DNA integrity in human spermatozoa: relationships with semen quality. J Androl. 2000;21:33–44.PubMedGoogle Scholar
  33. 33.
    Ochsendorf FR, Thiele J, Fuchs J, et al. Chemiluminescence in semen of infertile men. Andrologia. 1994;26:289–93.PubMedGoogle Scholar
  34. 34.
    Agarwal A, Prabakaran S, Allamaneni S. What an andrologist/urologist should know about free radicals and why. Urology. 2006;67:2–8.PubMedGoogle Scholar
  35. 35.
    Jones R, Mann T, Sherins R. Peroxidative breakdown of phospholipids in human spermatozoa, spermicidal properties of fatty acid peroxides, and protective action of seminal plasma. Fert Steril. 1979;31:531–7.Google Scholar
  36. 36.
    de Lamirande E, Gagnon C. Reactive oxygen species and human spermatozoa. II Depletion of adenosine triphosphate plays an important role in the inhibition of sperm motility. J Androl. 1992;13:379–86.PubMedGoogle Scholar
  37. 37.
    Whittington K, Harrison SC, Williams KM, et al. Reactive oxygen species (ROS) production and the outcome of diagnostic tests of sperm function. Int J Androl. 1999;22:236–42.PubMedGoogle Scholar
  38. 38.
    Kao SH, Chao HT, Chen HW, et al. Increase of oxidative stress in human sperm with lower motility. Fertil Steril. 2008;89:1183–90.PubMedGoogle Scholar
  39. 39.
    Dadoune JP, Mayaux MJ, Guihard-Moscato ML. Correlation between defects in chromatin condensation of human spermatozoa stained by aniline blue and semen characteristics. Andrologia. 1988;20:211–7.PubMedGoogle Scholar
  40. 40.
    Foresta C, Zorzi M, Rossato M, et al. Sperm nuclear instability and staining with aniline blue: abnormal persistence of histones in spermatozoa in infertile men. Int J Androl. 1992;15:330–7.PubMedGoogle Scholar
  41. 41.
    Oliva R. Protamines and male infertility. Hum Reprod Update. 2006;12:417–35.PubMedGoogle Scholar
  42. 42.
    Kodama H, Yamaguchi R, Fukuda J, et al. Increased oxidative deoxyribonucleic acid damage in the spermatozoa of infertile male patients. Fertil Steril. 1997;68:519–24.PubMedGoogle Scholar
  43. 43.
    Saleh RA, Agarwal A, Kandirali E, et al. Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertil Steril. 2002;78:1215–24.PubMedGoogle Scholar
  44. 44.
    Oger I, Da Cruz C, Panteix G, et al. Evaluating human sperm DNA integrity: relationship between 8-hydroxydeoxyguanosine quantification and the sperm chromatin structure assay. Zygote. 2003;11:367–71.PubMedGoogle Scholar
  45. 45.
    Wang X, Sharma RK, Sikka SC, et al. Oxidative stress is associated with increased apoptosis leading to spermatozoa DNA damage in patients with male factor infertility. Fertil Steril. 2003;80:531–5.PubMedGoogle Scholar
  46. 46.
    Henkel R, Kierspel E, Stalf T, et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in nonleukocytospermic patients. Fertil Steril. 2005;83:635–42.PubMedGoogle Scholar
  47. 47.
    Cayli S, Jakab A, Ovari L, et al. Biochemical markers of sperm function: male fertility and sperm selection for ICSI. RBM Online. 2003;7:462–8.PubMedGoogle Scholar
  48. 48.
    Sati L, Ovari L, Bennett D, et al. Double probing of human spermatozoa for persistent histones, surplus cytoplasm, apoptosis and DNA fragmentation. RBM Online. 2008;16:570–9.PubMedGoogle Scholar
  49. 49.
    Allen JW, Dix DJ, Collins BW, et al. HSP70-2 is part of the synaptonemal complex in mouse and hamster spermatocytes. Chromosoma. 1996;104:414–21.PubMedGoogle Scholar
  50. 50.
    Eddy EM. Role of heat shock protein HSP70-2 in spermatogenesis. Rev Reprod. 1999;4:23–30.PubMedGoogle Scholar
  51. 51.
    Tsunekawa N, Matsumoto M, Tone S, et al. The Hsp70 homolog gene, Hsc70t, is expressed under translational control during mouse spermiogenesis. Mol Reprod Dev. 1999;52:383–91.PubMedGoogle Scholar
  52. 52.
    Cayli S, Sakkas D, Vigue L, et al. Cellular maturity and apoptosis in human sperm: creatine kinase, caspase-3 and Bcl-XL levels in mature and diminished maturity sperm. Mol Hum Reprod. 2004;10:365–72.PubMedGoogle Scholar
  53. 53.
    Aitken RJ, Baker MA, Sawyer D. Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease. RBM Online. 2003;7:65–70.PubMedGoogle Scholar
  54. 54.
    Alvarez JG. DNA fragmentation in human spermatozoa: significance in the diagnosis and treatment of infertility. Minerva Ginecol. 2003;55:233–9.PubMedGoogle Scholar
  55. 55.
    Govin J, Caron C, Escoffier E, et al. Post-meiotic shifts in HSPA2/HSP70.2 chaperone activity during mouse spermatogenesis. J Biol Chem. 2006;281:37888–92.PubMedGoogle Scholar
  56. 56.
    Prinosilova P, Kruger T, Sati L, et al. Selectivity of hyaluronic acid binding for spermatozoa with normal Tygerberg strict morphology. RBM Online. 2009;18:177–83.PubMedGoogle Scholar
  57. 57.
    Bernardini L, Borini A, Preti S, et al. Study of aneuploidy in normal and abnormal germ cells from semen of fertile and infertile men. Hum Reprod. 1998;13:3406–13.PubMedGoogle Scholar
  58. 58.
    Colombero LT, Hariprashad JJ, Tsai MC, et al. Incidence of sperm aneuploidy in relation to semen characteristics and assisted reproductive outcome. Fertil Steril. 1999;72:90–6.PubMedGoogle Scholar
  59. 59.
    Calogero AE, De Palma A, Grazioso C, et al. Aneuploidy rate in spermatozoa of selected men with abnormal semen parameters. Hum Reprod. 2001;16:1172–9.PubMedGoogle Scholar
  60. 60.
    Templado C, Hoang T, Greene C, et al. Aneuploid spermatozoa in infertile men: teratozoospermia. Mol Reprod Dev. 2002;61:200–4.PubMedGoogle Scholar
  61. 61.
    Aran B, Vidal F, Vendrell JM, et al. Outcome of intracytoplasmic sperm injection in relation to the meiotic pattern in patients with severe oligoasthenozoospermia. Fertil Steril. 2003;80:91–5.PubMedGoogle Scholar
  62. 62.
    Pang MG, Kim YJ, Lee SH, et al. The high incidence of meiotic errors increases with decreased sperm count in severe male factor infertilities. Hum Reprod. 2005;20:1688–94.PubMedGoogle Scholar
  63. 63.
    Lee JD, Kamiguchi Y, Yanagimachi R. Analysis of chromosome constitution of human spermatozoa with normal and aberrant head morphologies after injection into mouse oocytes. Hum Reprod. 1996;11:1942–6.PubMedGoogle Scholar
  64. 64.
    Yakin K, Kahraman S. Certain forms of morphological anomalies of spermatozoa may reflect chromosomal aneuploidies. Hum Reprod. 2001;16:1779–80.PubMedGoogle Scholar
  65. 65.
    Kovanci E, Kovacs T, Moretti E, et al. FISH assessment of aneuploidy frequencies in mature and immature human spermatozoa classified by the absence or presence of cytoplasmic retention. Hum Reprod. 2001;16:1209–17.PubMedGoogle Scholar
  66. 66.
    Egozcue S, Blanco J, Vidal F, et al. Diploid sperm and the origin of triploidy. Hum Reprod. 2002;17:5–7.PubMedGoogle Scholar
  67. 67.
    Celik-Ozenci C, Catalanotti J, Jakab A, et al. Human sperm maintain their shape following decondensation and denaturation for fluorescent in situ hybridization: shape analysis and objective morphometry. Biol Reprod. 2003;69:1347–55.PubMedGoogle Scholar
  68. 68.
    Celik-Ozenci C, Jakab A, Kovacs T, et al. Sperm selection for ICSI: shape properties do not predict the absence or presence of numerical chromosomal aberrations. Hum Reprod. 2004;19:2052–9.PubMedGoogle Scholar
  69. 69.
    Zavaczki Z, Celik-Ozenci C, Ovari L, et al. Dimensional assessment of X-bearing and Y-bearing haploid and disomic human sperm with the use of fluorescence in situ hybridization and objective morphometry. Fertil Steril. 2006;85:121–7.PubMedGoogle Scholar
  70. 70.
    Simpson JL, Lamb DJ. Genetic effects of intracytoplasmic sperm injection. Semin Reprod Med. 2001;19:239–49.PubMedGoogle Scholar
  71. 71.
    Van Steirteghem A, Bonduelle M, Liebaers I, et al. Children born after assisted reproductive technology. Am J Perinatol. 2002;19:59–65.PubMedGoogle Scholar
  72. 72.
    Barri PN. Multiple pregnancies: a plea for informed caution. Hum Reprod Update. 2005;11:1–2.PubMedGoogle Scholar
  73. 73.
    Bonduelle M, Wennerholm UB, Loft A, et al. A multi-centre cohort study of the physical health of 5-year-old children conceived after intracytoplasmic sperm injection, in vitro fertilization and natural conception. Hum Reprod. 2005;20:413–9.PubMedGoogle Scholar
  74. 74.
    Jakab A, Sakkas D, Delpiano E, et al. Intracytoplasmic sperm injection: a novel selection method for sperm with normal frequency of chromosomal aneuploidies. Fertil Steril. 2005;84:1665–73.PubMedGoogle Scholar
  75. 75.
    Bonduelle M, Van Assche E, Joris H, et al. Prenatal testing in ICSI pregnancies: incidence of chromosomal anomalies in 1586 karyotypes and relation to sperm parameters. Hum Reprod. 2002;17:2600–14.PubMedGoogle Scholar
  76. 76.
    Huszar G, Patrizio P, Vigue L, et al. Cytoplasmic extrusion and the switch from creatine kinase B to M isoform are completed by the commencement of epididymal transport in human and stallion spermatozoa. J Androl. 1998;19:11–20.PubMedGoogle Scholar
  77. 77.
    Seli E, Sakkas D. Spermatozoal nuclear determinants of reproductive outcome: implications for ART. Hum Reprod Update. 2005;11:337–49.PubMedGoogle Scholar
  78. 78.
    Borini A, Tarozzi N, Bizzaro D, et al. Sperm DNA fragmentation: paternal effect on early post-implantation embryo development in ART. Hum Reprod. 2006;21:2876–81.PubMedGoogle Scholar
  79. 79.
    Ochsendorf FR. Infections in the male genital tract and reactive oxygen species. Hum Reprod Update. 1999;5:399–420.PubMedGoogle Scholar
  80. 80.
    Li K, Shang X, Chen Y. High-performance liquid chromatographic detection of lipid peroxidation in human seminal plasma and its application to male infertility. Clin Chim Acta. 2004;346: 199–203.PubMedGoogle Scholar
  81. 81.
    Shang XJ, Li K, Ye ZQ, et al. Analysis of lipid peroxidative levels in seminal plasma of infertile men by high-performance liquid chromatography. Arch Androl. 2004;50:411–6.PubMedGoogle Scholar
  82. 82.
    Aitken RJ, Harkiss D, Buckingham D. Relationship between iron-catalysed lipid peroxidation potential and human sperm function. J Reprod Fertil. 1993;98:257–65.PubMedGoogle Scholar
  83. 83.
    Khosrowbeygi A, Zarghami N. Levels of oxidative stress biomarkers in seminal plasma and their relationship with seminal parameters. BMC Clin Pathol. 2007;7:6.PubMedGoogle Scholar
  84. 84.
    Aitken RJ, Wingate JK, De Iuliis GN, et al. Analysis of lipid peroxidation in human spermatozoa using BODIPY C11. Mol Hum Reprod. 2007;13:203–11.PubMedGoogle Scholar
  85. 85.
    Saleh RA, Agarwal A, Nada EA, et al. Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Fertil Steril. 2003;79:1597–605.PubMedGoogle Scholar
  86. 86.
    Simon L, Brunborg G, Stevenson M, Lutton D, McManus J, Lewis SE. Clinical significance of sperm DNA damage in assisted reproduction outcome. Human Reprod. 2010;25(7):1594–608.PubMedGoogle Scholar
  87. 87.
    Ozmen B, Koutlaki N, Youssry M, et al. DNA damage of human spermatozoa in assisted reproduction: origins, diagnosis, impacts and safety. RBM Online. 2007;14:384–95.PubMedGoogle Scholar
  88. 88.
    Nakamura H, Kimura T, Nakajima A, et al. Detection of oxidative stress in seminal plasma and fractionated sperm from subfertile male patients. Eur J Obstet Gynecol Reprod Biol. 2002;105: 155–60.PubMedGoogle Scholar
  89. 89.
    Loft S, Kold-Jensen T, Hjollund NH, et al. Oxidative DNA damage in human sperm influences time to pregnancy. Hum Reprod. 2003;18:1265–72.PubMedGoogle Scholar
  90. 90.
    Kobayashi H, Gil-Guzman E, Mahran AM, et al. Quality control of reactive oxygen species measurement by luminol-dependent chemiluminescence assay. J Androl. 2001;22:568–74.PubMedGoogle Scholar
  91. 91.
    Aitken RJ, Baker MA, O’Bryan M. Shedding light on chemiluminescence: the application of chemiluminescence in diagnostic andrology. J Androl. 2004;25:455–65.PubMedGoogle Scholar
  92. 92.
    Liochev SI, Fridovich I. Lucigenin (Bis-N-methylacridinium) as a mediator of superoxide anion production. Archives Biochem Biophys. 1997;337:115–20.Google Scholar
  93. 93.
    World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucous ınteraction. 4th ed. New York: Cambridge University; 1999.Google Scholar
  94. 94.
    Sharma RK, Pasqualotto FF, Nelson DR, et al. The reactive oxygen species-total antioxidant capacity score is a new measure of oxidative stress to predict male infertility. Hum Reprod. 1999;14:2801–7.PubMedGoogle Scholar
  95. 95.
    Said TM, Kattal N, Sharma RK, et al. Enhanced chemiluminescence assay vs colorimetric assay for measurement of the total antioxidant capacity of human seminal plasma. J Androl. 2003;24:676–80.PubMedGoogle Scholar
  96. 96.
    Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem. 2004;37:277–85.PubMedGoogle Scholar
  97. 97.
    Tremellen K. Oxidative stress and male infertility—a clinical perspective. Hum Reprod Update. 2008;14:243–58.PubMedGoogle Scholar
  98. 98.
    Smith GR, Kaune GH, Ch Parodi D, et al. Extent of sperm DNA damage in spermatozoa from men examined for infertility. Relationship with oxidative stress. Rev Med Chil. 2007;135: 279–86.PubMedGoogle Scholar
  99. 99.
    Yagci A, Murk W, Stronk J, et al. Spermatozoa bound to solid state hyaluronic acid show chromatin structure with high DNA chain ıntegrity: an Acridine Orange Fluorescence Study. J Androl. 2010;31:566–72.PubMedGoogle Scholar
  100. 100.
    Greco E, Scarselli F, Iacobelli M, et al. Efficient treatment of infertility due to sperm DNA damage by ICSI with testicular spermatozoa. Hum Reprod. 2005;20:226–30.PubMedGoogle Scholar
  101. 101.
    Greco E, Iacobelli M, Rienzi L, et al. Reduction of the incidence of sperm DNA fragmentation by oral antioxidant treatment. J Androl. 2005;26:349–53.PubMedGoogle Scholar
  102. 102.
    O’Connell M, McClure N, Lewis SE. Mitochondrial DNA deletions and nuclear DNA fragmentation in testicular and epididymal human sperm. Hum Reprod. 2002;17:1565–70.PubMedGoogle Scholar
  103. 103.
    Comhaire FH, El El Garem Y, Mahmoud A, et al. Combined conventional/antioxidant ‘Astaxanthin’ treatment for male infertility: a double blind, randomized trial. Asian J Androl. 2005;7:257–62.PubMedGoogle Scholar
  104. 104.
    Vicari E, Calogero AE. Effects of treatment with carnitines in infertile patients with prostato-vesiculo-epididymitis. Hum Reprod. 2001;16:2338–42.PubMedGoogle Scholar
  105. 105.
    Comhaire FH, Christophe AB, Zalata AA, et al. The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men. Prostaglandins Leukot Essent Fatty Acids. 2000;63:159–65.PubMedGoogle Scholar
  106. 106.
    Keskes-Ammar L, Feki-Chakroun N, Rebai T, et al. Sperm oxidative stress and the effect of an oral vitamin E and selenium supplement on semen quality in infertile men. Arch Androl. 2003;49:83–94.PubMedGoogle Scholar
  107. 107.
    Suleiman SA, Ali ME, Zaki ZM, et al. Lipid peroxidation and human sperm motility: protective role of vitamin E. J Androl. 1996;17:530–7.PubMedGoogle Scholar
  108. 108.
    Menezo YJ, Hazout A, Panteix G, et al. Antioxidants to reduce sperm DNA fragmentation: an unexpected adverse effect. RBM Online. 2007;14:418–21.PubMedGoogle Scholar
  109. 109.
    MacLeod J. The role of oxygen in the metabolism and motility of human spermatozoa. Am J Physiol. 1943;138:512–8.Google Scholar
  110. 110.
    Kobayashi T, Miyazaki T, Natori M, et al. Protective role of superoxide dismutase in human sperm motility: superoxide dismutase activity and lipid peroxide in human seminal plasma and spermatozoa. Hum Reprod. 1991;6:987–91.PubMedGoogle Scholar
  111. 111.
    Oeda T, Henkel R, Ohmori H, et al. Scavenging effect of N-acetyl-L-cysteine against reactive oxygen species in human semen: a possible therapeutic modality for male factor infertility? Andrologia. 1997;29:125–31.PubMedGoogle Scholar
  112. 112.
    Zheng RL, Zhang H. Effects of ferulic acid on fertile and ­asthenozoospermic infertile human sperm motility, viability, lipid peroxidation, and cyclic nucleotides. Free Radic Biol Med. 1997;22:581–6.PubMedGoogle Scholar
  113. 113.
    Donnelly ET, McClure N, Lewis SE. Glutathione and hypotaurine in vitro: effects on human sperm motility, DNA integrity and production of reactive oxygen species. Mutagenesis. 2000;15:61–8.PubMedGoogle Scholar
  114. 114.
    Rossi T, Mazzilli F, Delfino M, et al. Improved human sperm recovery using superoxide dismutase and catalase supplementation in semen cryopreservation procedure. Cell Tissue Bank. 2001;2:9–13.PubMedGoogle Scholar
  115. 115.
    Yenilmez E, Yildirmis S, Yulug E, et al. Ham’s F-10 medium and Ham’s F-10 medium plus vitamin E have protective effect against oxidative stress in human semen. Urology. 2006;67:384–7.PubMedGoogle Scholar
  116. 116.
    Lenzi A, Culasso F, Gandini L, et al. Placebo-controlled, double-blind, cross-over trial of glutathione therapy in male infertility. Hum Reprod. 1993;8:1657–62.PubMedGoogle Scholar
  117. 117.
    Lenzi A, Sgro P, Salacone P, et al. A placebo-controlled double-blind randomized trial of the use of combined l-carnitine and l-acetyl-carnitine treatment in men with asthenozoospermia. Fertil Steril. 2004;81:1578–84.PubMedGoogle Scholar
  118. 118.
    Scott R, MacPherson A, Yates RW, et al. The effect of oral selenium supplementation on human sperm motility. Br J Urol. 1998;82:76–80.PubMedGoogle Scholar
  119. 119.
    Balercia G, Regoli F, Armeni T, et al. Placebo-controlled double-blind randomized trial on the use of L-carnitine, L-acetylcarnitine, or combined L-carnitine and L-acetylcarnitine in men with idiopathic asthenozoospermia. Fertil Steril. 2005;84:662–71.PubMedGoogle Scholar
  120. 120.
    Agarwal A, Nallella KP, Allamaneni SS, et al. Role of antioxidants in treatment of male infertility: an overview of the literature. RBM Online. 2004;8:616–27.PubMedGoogle Scholar
  121. 121.
    Rolf C, Cooper TG, Yeung CH, et al. Antioxidant treatment of patients with asthenozoospermia or moderate oligoasthenozoospermia with high-dose vitamin C and vitamin E: a randomized, placebo-controlled, double-blind study. Hum Reprod. 1999;14:1028–33.PubMedGoogle Scholar
  122. 122.
    Tremellen K, Miari G, Froiland D, et al. A randomised control trial examining the effect of an antioxidant (Menevit) on pregnancy outcome during IVF-ICSI treatment. Aust N Z J Obstet Gynaecol. 2007;47:216–21.PubMedGoogle Scholar
  123. 123.
    Baker HW, Edgar D. Trials of antioxidants for male infertility. Aust N Z J Obstet Gynaecol. 2008;48:125–6.PubMedGoogle Scholar

Further Reading

  1. Gomez E, Buckingham DW, Brindle J, et al. Development of an image analysis system to monitor the retention of residual cytoplasm by human spermatozoa: correlation with biochemical markers of the cytoplasmic space, oxidative stress, and sperm function. J Androl. 1996;17:276–87.PubMedGoogle Scholar
  2. Pasqualotto FF, Sharma RK, Kobayashi H, et al. Oxidative stress in normospermic men undergoing infertility evaluation. J Androl. 2001;22:316–22.PubMedGoogle Scholar
  3. Garrido N, Meseguer M, Alvarez J, et al. Relationship among standard semen parameters, glutathione peroxidase/glutathione reductase activity, and mRNA expression and reduced glutathione content in ejaculated spermatozoa from fertile and infertile men. Fertil Steril. 2004;82:1059–66.PubMedGoogle Scholar
  4. Said TM, Agarwal A, Sharma RK, et al. Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility. Fertil Steril. 2004;82:871–7.PubMedGoogle Scholar
  5. Said TM, Agarwal A, Sharma RK, et al. Impact of sperm morphology on DNA damage caused by oxidative stress induced by beta-nicotinamide adenine dinucleotide phosphate. Fertil Steril. 2005;83: 95–103.PubMedGoogle Scholar
  6. Agarwal A, Sharma RK, Nallella KP, et al. Reactive oxygen species as an independent marker of male factor infertility. Fertil Steril. 2006;86:878–85.PubMedGoogle Scholar
  7. Sakkas D, Alvarez JG. Sperm DNA fragmentation: mechanism of origin, impact onreproductive outcome, and analysis. Fertil Steril. 2010; 93(4): 1027–1036.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Histology and Embryology, School of MedicineAkdeniz UniversityAntalyaTurkey
  2. 2.Sperm Physiology LaboratoryYale School of MedicineNew HavenUSA

Personalised recommendations