Evaluation of Post-Thaw Swim-Up Selection Combined With Glutathione Addition for Improvement of Sperm Chromatin Maturity in Normozoospermic Samples

Overview

Journal Cureus

Specialty Biomedical Engineering

Date 2023 Jul 14

PMID 37448405

Authors

Raghad Abd Albari Shimal

Amal Abdulwahid Mohammed

Essraa Mohsen Al-Essawe

Affiliations

Soon will be listed here.

Abstract

Background: Cryopreservation of human semen alters the spermatozoal structure, resulting in a reduction in sperm function parameters. Various antioxidants may be able to slow or prevent this type of injury. Glutathione (GSH) has numerous antioxidant properties; supplementing the semen with GSH before freezing may assist in the restoration of post-thaw sperm functionality.

Objective: To investigate the effect of adding 5mM of glutathione before freezing on human sperm cryosurvival.

Materials And Methods: Semen samples were collected from 30 patients (22 normozoospermic and 8 asthenozoospermic) after 3-5 days of sexual abstinence. Following liquefaction, macro- and microscopic examinations were performed. The samples were then divided into two equal aliquots: the first aliquot received 5 mM of glutathione before freezing, and the second aliquot was considered the control (without glutathione). The samples were frozen using the rapid cryopreservation method and then preserved for 7-10 days in a liquid nitrogen tank before being thawed. Thawed samples from each group were examined microscopically according to the WHO 2010 guidelines. Aniline blue was used to assess the maturity of the DNA. Then, the direct-swim-up technique was applied to thawed samples from both groups to select the best sperm quality.

Results: The cryopreservation process negatively impacts all sperm characteristics in both groups. Sperm DNA integrity decreased. Glutathione addition before freezing decreased sperm chromatin immaturity (SCI) percent compared to the control (22.98 ± 0.83, 20.79 ± 0.56). The post-thaw performance of the swim-up technique resulted in a select sperm population with high progressive motility (p = 0.04) and an improvement in DNA integrity in the treated group versus the control group after thawing.

Conclusion: The DNA integrity in normozoospermic samples was improved by adding 5 mM glutathione before freezing. Performing the swim-up technique after thawing had no effect on sperm quality in asthenozoospermic patients.

References

Le M, Nguyen T, Nguyen T, Nguyen V, Nguyen T, Nguyen V . Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects. Eur J Obstet Gynecol Reprod Biol. 2019; 234:14-20. DOI: 10.1016/j.ejogrb.2019.01.001. View

Zini A, Al-Hathal N . Antioxidant therapy in male infertility: fact or fiction?. Asian J Androl. 2011; 13(3):374-81. PMC: 3739339. DOI: 10.1038/aja.2010.182. View

Luberda Z . The role of glutathione in mammalian gametes. Reprod Biol. 2005; 5(1):5-17. View

Sabeti P, Pourmasumi S, Rahiminia T, Akyash F, Talebi A . Etiologies of sperm oxidative stress. Int J Reprod Biomed. 2016; 14(4):231-40. PMC: 4918773. View

Le M, Nguyen T, Nguyen T, Nguyen T, Nguyen T, Nguyen Q . Does conventional freezing affect sperm DNA fragmentation?. Clin Exp Reprod Med. 2019; 46(2):67-75. PMC: 6572667. DOI: 10.5653/cerm.2019.46.2.67. View

Eskiocak S, Gozen A, Yapar S, Tavas F, Kilic A, Eskiocak M . Glutathione and free sulphydryl content of seminal plasma in healthy medical students during and after exam stress. Hum Reprod. 2005; 20(9):2595-600. DOI: 10.1093/humrep/dei062. View

He F, Wang F, Yang Y, Yuan Z, Sun C, Zou H . The effect of growth hormone on the metabolome of follicular fluid in patients with diminished ovarian reserve. Reprod Biol Endocrinol. 2023; 21(1):21. PMC: 9969693. DOI: 10.1186/s12958-023-01073-x. View

Santonastaso M, Mottola F, Iovine C, Colacurci N, Rocco L . Protective Effects of Curcumin on the Outcome of Cryopreservation in Human Sperm. Reprod Sci. 2021; 28(10):2895-2905. PMC: 8523395. DOI: 10.1007/s43032-021-00572-9. View

Raijmakers M, Roelofs H, Steegers E, Mulder T, Knapen M, Wong W . Glutathione and glutathione S-transferases A1-1 and P1-1 in seminal plasma may play a role in protecting against oxidative damage to spermatozoa. Fertil Steril. 2003; 79(1):169-72. DOI: 10.1016/s0015-0282(02)04404-7. View

10.

Huang C, Tang Y, Hu J, Zhou W, Huang Z, Luo X . Update on techniques for cryopreservation of human spermatozoa. Asian J Androl. 2022; 24(6):563-569. PMC: 9809481. DOI: 10.4103/aja20229. View

11.

Chatterjee S, Gagnon C . Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev. 2001; 59(4):451-8. DOI: 10.1002/mrd.1052. View

12.

Opuwari C, Henkel R . An Update on Oxidative Damage to Spermatozoa and Oocytes. Biomed Res Int. 2016; 2016:9540142. PMC: 4749785. DOI: 10.1155/2016/9540142. View

13.

Estrada E, Rivera Del Alamo M, Rodriguez-Gil J, Yeste M . The addition of reduced glutathione to cryopreservation media induces changes in the structure of motile subpopulations of frozen-thawed boar sperm. Cryobiology. 2017; 78:56-64. DOI: 10.1016/j.cryobiol.2017.07.002. View

14.

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(4):745-756. DOI: 10.1007/s10561-016-9566-5. View

15.

Di Santo M, Tarozzi N, Nadalini M, Borini A . Human Sperm Cryopreservation: Update on Techniques, Effect on DNA Integrity, and Implications for ART. Adv Urol. 2011; 2012:854837. PMC: 3238352. DOI: 10.1155/2012/854837. View

16.

Rahiminia T, Hosseini A, Anvari M, Ghasemi-Esmailabad S, Talebi A . Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity. Taiwan J Obstet Gynecol. 2017; 56(4):472-476. DOI: 10.1016/j.tjog.2017.02.004. View

17.

Minaei M, Barbarestani M, Nekoonam S, Abdolvahabi M, Takzare N, Asadi M . Effect of Trolox addition to cryopreservation media on human sperm motility. Iran J Reprod Med. 2014; 10(2):99-104. PMC: 4163270. View

18.

Winkle T, Rosenbusch B, Gagsteiger F, Paiss T, Zoller N . The correlation between male age, sperm quality and sperm DNA fragmentation in 320 men attending a fertility center. J Assist Reprod Genet. 2008; 26(1):41-6. PMC: 2649333. DOI: 10.1007/s10815-008-9277-3. View

19.

Mangoli E, Talebi A, Anvari M, Taheri F, Vatanparast M, Rahiminia T . Vitamin C attenuates negative effects of vitrification on sperm parameters, chromatin quality, apoptosis and acrosome reaction in neat and prepared normozoospermic samples. Taiwan J Obstet Gynecol. 2018; 57(2):200-204. DOI: 10.1016/j.tjog.2018.02.006. View

20.

SHERMAN J . Synopsis of the use of frozen human semen since 1964: state of the art of human semen banking. Fertil Steril. 1973; 24(5):397-412. DOI: 10.1016/s0015-0282(16)39678-9. View