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The effect of temperature and storage time on DNA integrity after freeze-drying sperm from individuals with normozoospermia

  • Received : 2023.04.25
  • Accepted : 2023.08.28
  • Published : 2024.03.31

Abstract

Objective: This study evaluated the effects of temperature and storage time on the quality and DNA integrity of freeze-dried sperm from individuals with normozoospermia. Methods: Normal sperm samples from 15 men aged 24 to 40 years were studied. Each sample was divided into six groups: fresh, freezing (frozen in liquid nitrogen), freeze-dried then preserved at room temperature for 1 month (FD-1m-RT), freeze-dried then preserved at room temperature for 2 months (FD-2m-RT), freeze-dried then preserved at 4 ℃ for 1 month (FD-1m-4 ℃), and freeze-dried then preserved at 4 ℃ for 2 months (FD-2m-4 ℃). The morphology, progressive motility, vitality, and DNA integrity of the sperm were evaluated in all groups. Results: In all freeze-dried groups, sperm cells were immotile after rehydration. The freeze-dried groups also showed significantly less sperm vitality than the fresh and frozen groups. Significantly more morphological sperm abnormalities were found in the freeze-dried groups, but freeze-drying did not lead to a significantly higher DNA fragmentation index (DFI). The DFI was significantly higher in the FD-2m-RT group than in the other freeze-dried groups. Conclusion: The freeze-drying method preserved the integrity of sperm DNA. The temperature and duration of storage were also identified as factors that influenced the DFI. Accordingly, more research is needed on ways to improve sperm quality in the freeze-drying process.

Keywords

Acknowledgement

The authors would like to thank Iran University of Medical Science, Tehran, Iran, for their cooperation throughout the period of study.

References

  1. Amidi F, Pazhohan A, Shabani Nashtaei M, Khodarahmian M, Nekoonam S. The role of antioxidants in sperm freezing: a review. Cell Tissue Bank 2016;17:745-56. https://doi.org/10.1007/s10561-016-9566-5
  2. Johnson LA, Weitze KF, Fiser P, Maxwell WM. Storage of boar semen. Anim Reprod Sci 2000;62:143-72. https://doi.org/10.1016/S0378-4320(00)00157-3
  3. Bailey JL, Lessard C, Jacques J, Breque C, Dobrinski I, Zeng W, et al. Cryopreservation of boar semen and its future importance to the industry. Theriogenology 2008;70:1251-9. https://doi.org/10.1016/j.theriogenology.2008.06.014
  4. Lusignan MF, Li X, Herrero B, Delbes G, Chan PT. Effects of different cryopreservation methods on DNA integrity and sperm chromatin quality in men. Andrology 2018;6:829-35. https://doi.org/10.1111/andr.12529
  5. Abolghasemi H, Aghaiipour M, Nikougoftar M, Amirizadeh N, Mohammadi MT, Rahmani S, et al. Leukoreduction in packed cells filtered by home-made. Sci J Iran Blood Transfus Organ 2009;6:13-20.
  6. Gil L, Olaciregui M, Luno V, Malo C, Gonzalez N, Martinez F. Current status of freeze-drying technology to preserve domestic animals sperm. Reprod Domest Anim 2014;49 Suppl 4:72-81. https://doi.org/10.1111/rda.12396
  7. Wakayama T, Yanagimachi R. Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat Biotechnol 1998;16:639-41. https://doi.org/10.1038/nbt0798-639
  8. Arav A, Saragusty J. Directional freezing of sperm and associated derived technologies. Anim Reprod Sci 2016;169:6-13. https://doi.org/10.1016/j.anireprosci.2016.02.007
  9. Olaciregui M, Luno V, Gonzalez N, Domingo P, de Blas I, Gil L. Chelating agents in combination with rosmarinic acid for boar sperm freeze-drying. Reprod Biol 2017;17:193-8. https://doi.org/10.1016/j.repbio.2017.05.001
  10. Polge C, Smith AU, Parkes AS. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature 1949;164:666.
  11. Kimura Y, Yanagimachi R. Intracytoplasmic sperm injection in the mouse. Biol Reprod 1995;52:709-20. https://doi.org/10.1095/biolreprod52.4.709
  12. Asgari F, Gavahi A, Karimi M, Vatannejad A, Amjadi F, Aflatoonian R, et al. Risk of embryo aneuploidy is affected by the increase in sperm DNA damage in recurrent implantation failure patients under ICSI-CGH array cycles. Hum Fertil (Camb) 2022;25:872-80. https://doi.org/10.1080/14647273.2021.1920054
  13. Abdel Fattah SM, Mohmed HK, Mohamed MA. The potential protective effect of ferulic acid against gamma irradiation induced ovarian failure in rats. Egypt J Radiat Sci Appl 2019;32:1-12.
  14. Kashani MH, Ramezani M, Piravar Z. The effect of acrylamide on sperm oxidative stress, total antioxidant levels, tyrosine phosphorylation, and carboxymethyl-lysine expression: a laboratory study. Int J Reprod Biomed 2021;19:625-36.
  15. Cheraghi E, Sajadi SM, Soleimani Mehranjani M. The effect of quercetin on the quality of sperm parameters in frozen-thawed semen of patients with Asthenospermia. Andrologia 2021;53: e14167.
  16. Kawase Y, Wada NA, Jishage K. Evaluation of DNA fragmentation of freeze-dried mouse sperm using a modified sperm chromatin structure assay. Theriogenology 2009;72:1047-53. https://doi.org/10.1016/j.theriogenology.2009.06.021
  17. Shahmoradi E, Baheiraei N, Halvaei I. Trehalose attenuates detrimental effects of freeze-drying on human sperm parameters. Biopreserv Biobank 2022;20:31-7. https://doi.org/10.1089/bio.2020.0167
  18. Olaciregui M, Luno V, Gonzalez N, De Blas I, Gil L. Freeze-dried dog sperm: dynamics of DNA integrity. Cryobiology 2015;71:286-90. https://doi.org/10.1016/j.cryobiol.2015.08.001
  19. Bossi RL, Cabral M, Oliveira M, Lopes S, Hurtado R, Sampaio M, et al. Ultrastructural analysis of lyophilized human spermatozoa. JBRA Assist Reprod 2021;25:473-9. https://doi.org/10.5935/1518-0557.20210028
  20. Kaneko T, Nakagata N. Relation between storage temperature and fertilizing ability of freeze-dried mouse spermatozoa. Comp Med 2005;55:140-4.
  21. Olaciregui M, Gil L. Freeze-dried spermatozoa: a future tool? Reprod Domest Anim 2017;52 Suppl 2:248-54. https://doi.org/10.1111/rda.12838
  22. Gianaroli L, Magli MC, Stanghellini I, Crippa A, Crivello AM, Pescatori ES, et al. DNA integrity is maintained after freeze-drying of human spermatozoa. Fertil Steril 2012;97:1067-73. https://doi.org/10.1016/j.fertnstert.2012.02.014
  23. Anzalone DA, Palazzese L, Iuso D, Martino G, Loi P. Freeze-dried spermatozoa: an alternative biobanking option for endangered species. Anim Reprod Sci 2018;190:85-93. https://doi.org/10.1016/j.anireprosci.2018.01.010