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Correlation between sperm motility and sperm chromatin/DNA damage before and after cryopreservation and the effect of folic acid and nicotinic acid on post-thaw sperm quality in normozoospermic men

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Abstract

Cryopreservation exposes sperm to physical and chemical stresses causing cell damages and impairs sperm functions. The aim of this study was to evaluate the association between motility and sperm chromatin/DNA damage before and after cryopreservation and investigate the effects of folic acid and nicotinic acid on post-thaw sperm quality. Thirty semen samples were obtained from 30 normozoospermic men, aged between 25 and 45 years old. Each sample were divided into five aliquots to form the following groups: fresh, cryopreserved with sperm-freeze only (control), with nicotinic acid (10 mM), with folic acid (50 nM), and with a combination of folic acid (50 nM) + nicotinic acid (10 mM). Sperm viability and motility in each group were assessed by eosin-nigrosine staining and computer-aided sperm analysis respectively. Sperm chromatin quality was studied by aniline blue, toluidine blue, acridine orange staining methods and sperm chromatin dispersion test. Cryopreservation led to a significant reduction in sperm quality in comparison to fresh sample groups (p < 0.05). Sperm chromatin damage was negatively correlated with the percentage of progressively motile cells. Supplementation of the cryopreservation medium with folic acid or nicotinic acid induced a significant improvement in sperm parameters and chromatin quality, compared to control groups (p < 0.05). Meanwhile, the combination of folic acid + nicotinic acid showed a significant protective effect in post thaw sperm. In conclusion, cryopreservation generated oxidative stress, inducingsperm cryodamage, reducing progressive motility and sperm quality, as an indicator of significant chromatin/DNA damage. Folic acid and nicotinic acid exhibited a potential cryoprotective effect by enhancing sperm quality.

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References

  • Agarwal A, Durairajanayagam D, Du Plessis SS (2014) Utility of antioxidants during assisted reproductive techniques: an evidence-based review. Reprod Biol Endocrinol 12:112

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Amidi F, Pazhohan A, Nashtaei MS, Khodarahmian M, Nekoonam S (2016) The role of antioxidants in sperm freezing: a review. Cell Tissue Bank 17:745–756

    Article  CAS  PubMed  Google Scholar 

  • Anghel A, Zamfirescu S, Dragomir C, Nadolu D, Elena S, Florica B (2010) The effects of antioxidants on the cytological parameters of cryopreserved buck semen. Rom Biotechnol Lett 15:26–32

    CAS  Google Scholar 

  • Banihani S, Agarwal A, Sharma R, Bayachou M (2014) Cryoprotective effect of L-carnitine on motility, vitality and DNA oxidation of human spermatozoa. Andrologia 46:637–641

    Article  CAS  PubMed  Google Scholar 

  • Bateni Z, Azadi L, Tavalaee M, Kiani-Esfahani A, Fazilati M, Nasr-Esfahani MH (2014) Addition of Tempol in semen cryopreservation medium improves the post-thaw sperm function. Syst Biol Reprod Med 60:245–250

    Article  CAS  PubMed  Google Scholar 

  • Bühler S et al (2017) Effects of energy supply and nicotinic acid supplementation on phagocytosis and ROS production of blood immune cells of periparturient primi-and pluriparous dairy cows. Res Vet Sci 116:62–71. https://doi.org/10.1016/j.rvsc.2017.09.012

    Article  CAS  PubMed  Google Scholar 

  • Cooper TG et al (2010) World Health Organization reference values for human semen characteristics. Hum Reprod Update 16:231–245

    Article  PubMed  Google Scholar 

  • Crott JW, Mashiyama ST, Ames BN, Fenech M (2001) The effect of folic acid deficiency and MTHFR C677T polymorphism on chromosome damage in human lymphocytes in vitro. Cancer Epidemiol Prevent Biomark 10:1089–1096

    CAS  Google Scholar 

  • Duthie SJ (2011) Folate and cancer: how DNA damage, repair and methylation impact on colon carcinogenesis. J Inherit Metab Dis 34:101–109

    Article  CAS  PubMed  Google Scholar 

  • Duthie SJ, Hawdon A (1998) DNA instability (strand breakage, uracil misincorporation, and defective repair) is increased by folic acid depletion in human lymphocytes in vitro. FASEB J 12:1491–1497

    Article  CAS  PubMed  Google Scholar 

  • Erenpreiss J, Bars J, Lipatnikova V, Erenpreisa J (2001) Comparative study of cytochemical tests for sperm chromatin integrity. J Androl 22:45–53

    CAS  PubMed  Google Scholar 

  • Fleury C, Mignotte B, Vayssière J-L (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84:131–141

    Article  CAS  PubMed  Google Scholar 

  • Gadea J, Molla M, Selles E, Marco M, Garcia-Vazquez F, Gardon J (2011) Reduced glutathione content in human sperm is decreased after cryopreservation: effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology 62:40–46

    Article  CAS  PubMed  Google Scholar 

  • Gharagozloo P, Aitken RJ (2011) The role of sperm oxidative stress in male infertility and the significance of oral antioxidant therapy. Hum Reprod 26:1628–1640

    Article  PubMed  Google Scholar 

  • Ghasemi N, Dashti G-R, Amoozgar F, Vaez S-A (2014) Effect of cholesterol, iron and vitamin E on protamine deficiency and DNA fragmentation of male rabbit sperm. J Isfahan Med Sch 31(259):1769–1778

    Google Scholar 

  • Gianaroli L, Magli MC, Stanghellini I, Crippa A, Crivello AM, Pescatori ES, Ferraretti AP (2012) DNA integrity is maintained after freeze-drying of human spermatozoa. Fertil Steril 97(1067–1073):e1061

    Google Scholar 

  • Giwercman A, Richthoff J, Hjøllund H, Bonde JP, Jepson K, Frohm B, Spano M (2003) Correlation between sperm motility and sperm chromatin structure assay parameters. Fertil Steril 80:1404–1412

    Article  PubMed  Google Scholar 

  • Greenberg JA, Bell SJ, Guan Y, Yu Y-H (2011) Folic acid supplementation and pregnancy: more than just neural tube defect prevention. Rev Obstet Gynecol 4:52–59

    PubMed  PubMed Central  Google Scholar 

  • Iranpour FG, Fazelian K, Dashti GR (2017) Thymoquinone as a natural spermostatic substance in reproductive medicine: an experimental study. Int J Reprod Biomed 15:641

    Article  CAS  Google Scholar 

  • Irvine DS, Twigg JP, Gordon EL, Fulton N, Milne PA, Aitken RJ (2000) DNA integrity in human spermatozoa: relationships with semen quality. J Androl 21:33–44

    CAS  PubMed  Google Scholar 

  • Joshi R, Adhikari S, Patro B, Chattopadhyay S, Mukherjee T (2001) Free radical scavenging behavior of folic acid: evidence for possible antioxidant activity. Free Radic Biol Med 30:1390–1399

    Article  CAS  PubMed  Google Scholar 

  • Kamen B (1997) Folate and antifolate pharmacology. Semin Oncol 24:18–39

    Google Scholar 

  • Keshtgar S, Iravanpour F, Gharesi-Fard B, Kazerooni M (2016) Combined effect of Trolox and EDTA on frozen-thawed sperm quality. Iran J Med Sci 41:230

    PubMed  Google Scholar 

  • Khamsuk K, Sinawat S, Seejorn K, Pongsritasana T, Sukkasame S (2014) The effect of folic acid on post-thaw quality of human spermatozoa. Srinagarind Med J 29:141–145

    Google Scholar 

  • Kim Y-J et al (2015) Effect of nicotinic acid on sperm characteristic and oocyte development after in vitro fertilization using cryopreserved boar semen. J Embryo Transf 30:7–15

    Article  Google Scholar 

  • Kotdawala AP et al (2012) Addition of zinc to human ejaculate prior to cryopreservation prevents freeze–thaw-induced DNA damage and preserves sperm function. J Assist Reprod Genet 29:1447–1453

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee Y-J et al (2014) Effect of nicotinic acid on fresh semen characteristics in miniature pigs. J Embryo Transf 29:385–391

    Article  Google Scholar 

  • Lee Y-J et al (2016) Effects of L-carnitine and nicotinic acid on sperm characteristics in miniature pigs. Reprod Dev Biol 40:1–5

    Article  CAS  Google Scholar 

  • Murphy LE et al (2011) Folate and vitamin B12 in idiopathic male infertility Asian. J Androl 13:856

    CAS  Google Scholar 

  • Najafi A et al (2016) Supplementation of freezing and thawing media with brain-derived neurotrophic factor protects human sperm from freeze–thaw-induced damage. Fertil Steril 106(1658–1665):e1654

    Google Scholar 

  • Nakano E, Higgins JA, Powers HJ (2001) Folate protects against oxidative modification of human LDL. Br J Nutr 86:637–639

    Article  CAS  PubMed  Google Scholar 

  • Oberoi B, Kumar S, Talwar P (2014) Study of human sperm motility post-cryopreservation. Med J Armed Forces India 70:349–353

    Article  PubMed  PubMed Central  Google Scholar 

  • Oehninger S, Duru NK, Srisombut C, Morshedi M (2000) Assessment of sperm cryodamage and strategies to improve outcome. Mol Cell Endocrinol 169:3–10

    Article  CAS  PubMed  Google Scholar 

  • Peris SI, Morrier A, Dufour M, Bailey JL (2004) Cryopreservation of ram semen facilitates sperm DNA damage: the relationship between sperm andrological parameters and the sperm chromatin structure assay. J Androl 25:224–233

    Article  CAS  PubMed  Google Scholar 

  • Pietrzik K, Bailey L, Shane B (2010) Folic acid and L-5-methyltetrahydrofolate. Clin Pharmacokinet 49:535–548

    Article  CAS  PubMed  Google Scholar 

  • Pour H, Tahbasbi A, Naserain A-A (2013) The influence of vitamin E on semen characteristics of ghezel rams in during cooling and frozen process. Eur J Med Res 2:94–99

    CAS  Google Scholar 

  • Rahiminia T, Hosseini A, Anvari M, Ghasemi-Esmailabad S, Talebi AR (2017) Modern human sperm freezing: effect on DNA, chromatin and acrosome integrity Taiwanese. J Obstet Gynecol 56:472–476

    Google Scholar 

  • Taghizabet N, Mangoli E, Anbari F, Masoodi SA, Talebi AR, Mazrooei M (2016) The effect of heracleum persicum (Golpar) oil and alcoholic extracts on sperm parameters and chromatin quality in mice. Int J Reprod Biomed 14:365

    Article  CAS  Google Scholar 

  • Tejada RI, Mitchell JC, Norman A, Marik JJ, Friedman S (1984) A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril 42:87–91

    Article  CAS  PubMed  Google Scholar 

  • Toghiani S, Dashti GR, Roudbari NH, Roozbehani S (2016) Effect of ascorbic acid and menthone on the caspase 3 in the sperm cells of acyclovir treated rats. Acta Medica 32:1213

    Google Scholar 

  • Verdin E, Hirschey MD, Finley LW, Haigis MC (2010) Sirtuin regulation of mitochondria: energy production, apoptosis, and signaling. Trends Biochem Sci 35:669–675

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Verza S Jr, Feijo CM, Esteves SC (2009) Resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing. Int Braz J Urol 35:581–591

    Article  PubMed  Google Scholar 

  • Zribi N et al (2012) Effect of freezing–thawing process and quercetin on human sperm survival and DNA integrity. Cryobiology 65:326–331

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was funded by Isfahan University of Medical Sciences, Isfahan, Iran. The authors would like to thank Mrs. Fatemeh Amozegar and Mrs. Maryam Aliakbary for their technical support. The authors would like to thank the participating and express their gratitude to Isfahan University of medical sciences for its financial support.

Funding

This study was funded by Isfahan University of Medical Sciences, Isfahan, Iran (Grant No. 396378).

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Authors and Affiliations

  1. Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Fahimeh Zamani Rarani, Farhad Golshan-Iranpour & Gholam Reza Dashti

  2. Saint Maryam Fertility and Infertility Center, Shahid Beheshti Hospital, Isfahan, Iran

    Farhad Golshan-Iranpour & Gholam Reza Dashti

Authors
  1. Fahimeh Zamani Rarani
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  2. Farhad Golshan-Iranpour
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  3. Gholam Reza Dashti
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Correspondence to Gholam Reza Dashti.

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The procedures in this studies involving human participants was performed 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.

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Rarani, F.Z., Golshan-Iranpour, F. & Dashti, G.R. Correlation between sperm motility and sperm chromatin/DNA damage before and after cryopreservation and the effect of folic acid and nicotinic acid on post-thaw sperm quality in normozoospermic men. Cell Tissue Bank 20, 367–378 (2019). https://doi.org/10.1007/s10561-019-09775-6

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