In vitro Alpha Lipoic Acid supplementation in freeze-dried human sperm: the impact on DNA fragmentation index

Rezia Octarina, M. P. B. Dyah Pramesti, Agustinus Agustinus

Abstract

Freeze-drying technique is an alternative method on sperm preservation. However, this process can results in DNA damage. Adding antioxidants before freezing can reduce the detrimental effects of ROS on spermatozoa. Alpha lipoic acid (ALA) is one of the potent antioxidant that can be used to protect the sperm DNA. This study was aimed to determine the effect of ALA supplementation before freeze-drying on the DNA fragmentation index (DFI) of human spermatozoa. Nine semen samples were collected and evaluated for DFI (O1) then divided into 3 groups; freeze-drying without ALA (O2), freeze-drying with ALA 1,25 mg (O3) and freeze-drying with ALA 2,5 mg (O4).All three groups were stored at 4oC for 1 week. Sperm DFI were evaluated before and after freeze-drying. The result of this study showed a significant increase in sperm DFI in all three groups after freeze-drying (p<0,05) when compared to DFI before freeze-drying (14,67 ±3,295). However there was no significant difference between O2 (26,90 ±9,180) and O4 (29,29 ±5,524) group where the supplementation of 2,5 mg ALA did not have significant effect on protecting the DNA of freeze-dried human spermatozoa. The highest DFI was in the O3 group (44,64 ±11,717). Therefore, ALA supplementation before freeze-drying does not have a significant effect on protecting the DNA of freeze-dried human spermatozoa.

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References

Buanayuda, G. W., Lunardhi, H., & Mansur, I. G. (2019). Effect of in-vitro alpha lipoic acid addition on spermatozoa motility in sperm preparation process. Fol Med Indones, 55(4), 246-50. http://dx.doi.org/10.20473/fmi.v55i4.17309

Canepa, P., Dal Lago, A., De Leo, C., Gallo, M., Rizzo, C., Licata, E., Anserini, P., Rago, R., & Scaruffi, P. (2018). Combined treatment with myo-inositol, alpha-lipoic acid, folic acid and vitamins significantly improves sperm parameters of sub-fertile men: A multi-centric study. Eur Rev Med Pharmacol Sci, 22, 7078-85.

Donnelly, E. T., McClure, N., & Lewis, S. E. M. (1999). The effect of ascorbate and α-tocopherol supplementation in vitro on DNA integrity and hydrogen peroxide-induced DNA damage in human spermatozoa. Mutagenesis, 14(5), 505-11.

El Barky, A. R., Hussein, S. A., & Mohamed, T. M. (2017). The potent antioxidant alpha lipoic acid. J Plant Chem and Ecophysiol, 2(1), 1-5.

Gianaroli, L., Magli, M. C., Stanghellini, I., Crippa, A., Crivello, A. M., Pescatori, E. S., & Ferraretti, A. P. (2012). DNA integrity is maintained after freeze-drying of human spermatozoa. Fertility and Sterility, 97(5), 1067-1073.e1. https://doi.org/10.1016/j.fertnstert.2012.02.014

Gil, L., Olaciregui, M., Luño, V., Malo, C., González, N., & Martínez, F. (2014). Current Status of Freeze-Drying Technology to Preserve Domestic Animals Sperm. Reproduction in Domestic Animals, 49, 72–81. https://doi.org/10.1111/rda.12396

Haghighian, H. K., Haidari, F., Mohammadi-asl, J., & Dadfar, M. (2015). Randomized, triple-blind, placebo-controlled clinical trial examining the effects of alpha-lipoic acid supplement on the spermatogram and seminal oxidative stress in infertile men. Fertility and Sterility, 104(2), 318–324. https://doi.org/10.1016/j.fertnstert.2015.05.014

Hu, J.-H., Sun, X.-Z., Li, Q.-W., Zhang, T., Hu, X.-C., Hu, J.-H., & Wang, L.-Q. (2013). The effect of Laminaria japonic polysaccharide on sperm characteristics and biochemical parameters in cryopreserved boar sperm. Animal Reproduction Science, 139(1–4), 95–100. https://doi.org/10.1016/j.anireprosci.2013.03.015

Ibrahim, S. F., Osman, K., Das, S., Othman, A. M., Majid, N. A., & Rahman, M. P. A. (2008). A study of the antioxidant effect of alpha lipoic acids on sperm quality. Clinics, 63(4), 545–550. https://doi.org/10.1590/S1807-59322008000400022

Kusakabe, H. (2011). Chromosomal integrity and DNA damage in freeze-dried spermatozoa. Reproductive Medicine and Biology, 10(4), 199–210. https://doi.org/10.1007/s12522-011-0092-7

Lestari, S. W., Lestari, S. H., & Pujianto, D. A. (2018). Sperm quality after swim up and density gradient centrifugation sperm preparation with supplementation of alpha lipoic acid (ALA): A preliminary study. AIP Conference Proceedings 1933, 030015-1-6. https://doi.org/10.1063/1.5023962

Luño, V., Gil, L., Olaciregui, M., González, N., Jerez, R. A., & de Blas, I. (2014). Rosmarinic acid improves function and in vitro fertilising ability of boar sperm after cryopreservation. Cryobiology, 69(1), 157–162. https://doi.org/10.1016/j.cryobiol.2014.07.002

Olaciregui, M., & Gil, L. (2017). Freeze-dried spermatozoa: A future tool? Reproduction in Domestic Animals, 52, 248–254. https://doi.org/10.1111/rda.12838

Patrizio, P., Natan, Y., Barak, Y., Levi Setti, P., & Arav, A. (2016). A simple new method for the freeze-drying and storage of human sperm. Fertility and Sterility, 106(3), e307. https://doi.org/10.1016/j.fertnstert.2016.07.871

Peitao, W., Cuihua, S., Daqing, S., Li, Q., Liu, Z., & Xiuyan, F. (2008). The Effect of Freeze Drying on the Deoxyribonucleic Acid Chain of Human Sperm. 2008 2nd International Conference on Bioinformatics and Biomedical Engineering, 919–923. https://doi.org/10.1109/ICBBE.2008.226

Pramesti, M. P. B. D., Lunardhi, H., & I’tishom, R. (2010). Human sperm preservation using freeze-drying an experimental laboratory study. Jurnal Kesehatan Reproduksi, 1(1), 14-22.

Saili, T., Prasetyaningtyas, W. E., Setiadi, M. A., Agungpriyono, S., Agungpriyono, S., & Boediono, A. (2006). Status DNA spermatozoa domba setelah proses pengeringbekuan. JITV, 11(3), 215-21.

Shen, T., Jiang, Z.-L., Li, C.-J., Hu, X.-C., & Li, Q.-W. (2016). Effect of alpha-lipoic acid on boar spermatozoa quality during freezing–thawing. Zygote, 24(2), 259–265. https://doi.org/10.1017/S0967199415000155

Tibullo, D., Li Volti, G., Giallongo, C., Grasso, S., Tomassoni, D., Anfuso, C. D., Lupo, G., Amenta, F., Avola, R., & Bramanti, V. (2017). Biochemical and clinical relevance of alpha lipoic acid: Antioxidant and anti-inflammatory activity, molecular pathways and therapeutic potential. Inflammation Research, 66(11), 947–959. https://doi.org/10.1007/s00011-017-1079-6

World Health Organization (Ed.). (2010). WHO laboratory manual for the examination and processing of human semen (5th ed). World Health Organization.

Yeni, D., Fidan, A. F., Ciğerci, I. H., Konuk, M., Avdatek, F., & Gündoğan, M. (2012). Effect of α-lipoic acid on sperm quality, reproductive tract measures in thinner exposed rats: α-lipoic acid sperm quality thinner induced rats. Andrologia, 44, 74–80. https://doi.org/10.1111/j.1439-0272.2010.01140.x

Zhu, W., Li, J., & Xiao, L. (2016). Changes on membrane integrity and ultrastructure of human sperm after freeze-drying. Journal of Reproduction & Contraception, 27(2), 76-81. https://doi.org/10.7669/j.issn.1001-7844.2016.02.0076


DOI: http://dx.doi.org/10.30651/jqm.v5i1.4374