https://doi.org/10.4081/aiua.2024.12525

Pentoxifylline treatment as a safe method for selecting viable testicular spermatozoa before cryopreservation of a small numbers of spermatozoa in azoospermia individuals

Vol. 96 No. 2 (2024)
Submitted: March 29, 2024
Accepted: April 13, 2024
Published: June 27, 2024
Non-obstructive azoospermia, pentoxifylline;, testicular sperm, single sperm cryopreservation
Abstract Views: 741
PDF: 292
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Authors

Keivan Lorian
International Campus, Shahid Sadoughi University of Medical Science; Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran, Islamic Republic of.
Serajoddin Vahidi
Andrology Research center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran, Islamic Republic of.
Fatemeh Dehghanpour
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran, Islamic Republic of.
Fatemeh Anbari
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran, Islamic Republic of.
Azam Agha-Rahimi
63rahimi@gmail.com
https://orcid.org/0000-0002-7579-6192
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran, Islamic Republic of.

Background: Single sperm cryopreservation (SSC) is a specific technique especially used in individuals with small numbers of sperm who suffered from non-obstructive azoospermia (NOA). Testicular specimens possess poor motility and low population of viable spermatozoa. Therefore, sperm selection methods such as applying pentoxifylline (PTX) may improve motility in these cases. The main aim of this study was to evaluate the protective effects of PTX on testicular spermatozoa before and after performing SSC.
Methods: Thirty testicular samples were obtained from men with azoospermia. This study was conducted in two phases. Phase 1 evaluated the effect of PTX for sperm selection before SSC. Twenty testicular samples were divided to two experimental groups: SSC without (I) and with PTX treatment (II). For PTX treatment spermatozoa were incubated with PTX at 37°C for 30 min and only motile spermatozoa were selected for SSC. In phase 2, ten testicular samples were cryopreserved with SSC and warming procedure was carried out in droplet with and without PTX. Motility and viability rates, morphology by motile sperm organelle morphology examination (MSOME), DNA fragmentation by sperm chromatin dispersion test (SCD) and mitochondrial membrane potential (MMP) were evaluated.
Results: In phase 1, post warm motility rate was higher in PTX exposed group compared to the unexposed group (25.6 ± 8.13 vs. 0.85 ± 2.1) (p > 0.00). Recovery rate, viability and morphology were not significantly different between groups. DNA integrity and MMP were also similar between both groups. In phase 2 although motility increased in PTX group compared to without PTX group (29.30 ± 12.73 vs. 1.90 ± 2.64) (p > 0.00), the viability rate was not different (70.40 ± 12.12 vs. 65.30 ± 11.87). All above mentioned parameters were similar between the two SSC groups.
Conclusions: Supplementation of testicular spermatozoa with PTX before cryopreservation increases motility and did not have adverse effects on viability, morphology, DNA integrity and MMP. PTX could be used as sperm selection method before single sperm cryopreservation, but PTX could not maintain motile the most of viable testicular sperms.

Dimensions

Altmetric

PlumX Metrics

Downloads

Download data is not yet available.

Citations

Crossref
Scopus
Google Scholar
Europe PMC
Cetinkaya M, Onem K, Zorba OU, et al. Evaluation of microdissection testicular sperm extraction results in patients with nonobstructive azoospermia: independent predictive factors and best cutoff values for sperm retrieval. Urology Journal. 2015; 12:2436-43.
Cocuzza M, Alvarenga C, Pagani R. The epidemiology and etiology of azoospermia. Clinics. 2013; 68:15-26. DOI: https://doi.org/10.6061/clinics/2013(Sup01)03
Modarresi T, Hosseinifar H, Hampa AD, et al. Predictive factors of successful microdissection testicular sperm extraction in patients with presumed sertoli cell-only syndrome. Int J Fertil Steril. 2015; 9:107-12.
AbdelHafez F, Bedaiwy M, El-Nashar SA, et al. Techniques for cryopreservation of individual or small numbers of human spermatozoa: a systematic review. Hum Reprod Update. 2009; 15:153-64. DOI: https://doi.org/10.1093/humupd/dmn061
Liu S, Li F. Cryopreservation of single-sperm: where are we today? Reprod Biol Endocrinol. 2020; 18:41. DOI: https://doi.org/10.1186/s12958-020-00607-x
Cohen J, Garrisi GJ, Congedo-Ferrara TA, et al. Cryopreservation of single human spermatozoa. Hum Reprod. 1997; 12:994-1001. DOI: https://doi.org/10.1093/humrep/12.5.994
Endo Y, Fujii Y, Motoyama H. Clinical and neonatal outcomes of individually vitrified human sperm with Cryotop and Cell Sleeper. Cryobiology. 2022; 108:78-81. DOI: https://doi.org/10.1016/j.cryobiol.2022.07.003
Karacan M, Alwaeely F, Erkan S, et al. Outcome of intracytoplasmic sperm injection cycles with fresh testicular spermatozoa obtained on the day of or the day before oocyte collection and with cryopreserved testicular sperm in patients with azoospermia. Fertil Steril. 2013; 100:975-80. DOI: https://doi.org/10.1016/j.fertnstert.2013.06.031
Verheyen G, Joris H, Crits K, et al. Comparison of different hypoosmotic swelling solutions to select viable immotile spermatozoa for potential use in intracytoplasmic sperm injection. Hum Reprod Update. 1997; 3:195-203. DOI: https://doi.org/10.1093/humupd/3.3.195
Gholizadeh L, Khalili MA, Maleki B, et al. Quality of testicular spermatozoa improves with changes in composition of culture medium. Basic Clin Androl. 2023; 33:22. DOI: https://doi.org/10.1186/s12610-023-00198-8
Luo Y, Peng F, Li L, et al. Sperm Activation with Pentoxifylline Is Beneficial For Non-Obstructive Azoospermia Patients Using Testicular Sperm before Intracytoplasmic Sperm Injection. Int J Women’s Health Care 2022; 7:73. DOI: https://doi.org/10.33140/IJWHC.07.02.02
Mahaldashtian M, Khalili MA, Nottola SA, et al. Does in vitro application of pentoxifylline have beneficial effects in assisted male reproduction? Andrologia. 2021; 53:e13722. DOI: https://doi.org/10.1111/and.13722
Esteves SC, Varghese AC. Laboratory handling of epididymal and testicular spermatozoa: what can be done to improve sperm injections outcome. J Hum Reprod Sci.. 2012; 5:233. DOI: https://doi.org/10.4103/0974-1208.106333
Bagheripour N, Khalili MA, Nabi A, et al. A new cryotop vial device system provides an aseptic cryoprotectant-free and centrifugefree cryopreservation of human spermatozoa (a closed system). Cryobiology. 2023; 111:70-75. DOI: https://doi.org/10.1016/j.cryobiol.2023.03.001
Maleki B, Khalili MA, Gholizadeh L, et al. Single sperm vitrification with permeable cryoprotectant-free medium is more effective in patients with severe oligozoospermia and azoospermia. Cryobiology. 2022; 104:15-22. DOI: https://doi.org/10.1016/j.cryobiol.2021.11.176
Mahaldashtian M, Khalili MA, Mangoli E, et al. Pentoxifylline treatment had no detrimental effect on sperm DNA integrity and clinical characteristics in cases with non-obstructive azoospermia. Zygote. 2023; 31:8-13. DOI: https://doi.org/10.1017/S0967199422000247
Sallam HN, Farrag A, Agameya A-F, et al. The use of the modified hypo-osmotic swelling test for the selection of immotile testicular spermatozoa in patients treated with ICSI: a randomized controlled study. Hum Reprod. 2005; 20:3435-40. DOI: https://doi.org/10.1093/humrep/dei249
Cassuto NG, Bouret D, Plouchart JM, et al. A new real-time morphology classification for human spermatozoa: a link for fertilization and improved embryo quality. Fertil Steril. 2009; 92:1616-25. DOI: https://doi.org/10.1016/j.fertnstert.2008.08.088
Nordhoff V. How to select immotile but viable spermatozoa on the day of intracytoplasmic sperm injection? An embryologist's view. Andrology. 2015; 3:156-62. DOI: https://doi.org/10.1111/andr.286
Xian Y, Jiang M, Liu B, et al. A cryoprotectant supplemented with pentoxifylline can improve the effect of freezing on the motility of human testicular sperm. Zygote. 2022; 30:92-7. DOI: https://doi.org/10.1017/S0967199421000368
Endo Y, Fujii Y, Shintani K, et al. Single spermatozoon freezing using Cryotop. Journal of Mammalian Ova Research. 2011; 28:47-52. DOI: https://doi.org/10.1274/jmor.28.47
Sun J, Chen W, Zhou L, et al. Successful delivery derived from cryopreserved rare human spermatozoa with novel cryopiece. Andrology. 2017; 5:832-7. DOI: https://doi.org/10.1111/andr.12380
Nabi A, Khalili MA, Talebi AR, et al. In-vitro application of pentoxifylline preserved ultrastructure of spermatozoa after vitrification in asthenozoospermic patients. Urology Journal. 2017; 14:4038-43.
Mahaldashtian M, Khalili MA, Vatanparast M, et al. The effect of pentoxifylline and calcium ionophore treatment on sperm cell biology in oligoasthenoteratozoospermia samples. Zygote. 2023; 31:85-90. DOI: https://doi.org/10.1017/S0967199422000582
Asokan Y, Honguntikar SD, Uppangala S, et al. In situ viability detection assays induce heat-shock protein 70 expression in spermatozoa without affecting the chromatin integrity. Andrologia. 2015; 47:958-65. DOI: https://doi.org/10.1111/and.12364
Nabi A, Khalili MA, Fesahat F, et al. Pentoxifylline increase sperm motility in devitrified spermatozoa from asthenozoospermic patient without damage chromatin and DNA integrity. Cryobiology. 2017; 76:59-64. DOI: https://doi.org/10.1016/j.cryobiol.2017.04.008
Boguenet M, Bouet P-E, Spiers A, et al. Mitochondria: their role in spermatozoa and in male infertility. Hum Reprod Update. 2021; 27:697-719. DOI: https://doi.org/10.1093/humupd/dmab001

How to Cite

Lorian, K., Vahidi, S., Dehghanpour, F., Anbari, F., & Agha-Rahimi, A. (2024). Pentoxifylline treatment as a safe method for selecting viable testicular spermatozoa before cryopreservation of a small numbers of spermatozoa in azoospermia individuals. Archivio Italiano Di Urologia E Andrologia, 96(2). https://doi.org/10.4081/aiua.2024.12525
Copyright (c) 2024 the Author(s) Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

PAGEPress has chosen to apply the Creative Commons Attribution NonCommercial 4.0 International License (CC BY-NC 4.0) to all manuscripts to be published.