The Effect of Edaravone Supplementation Prior to Cryopreservation on Sperm Parameters, DNA Integrity, Apoptosis, Lipid Peroxidation, and Mitochondrial Membrane Potential in Infertile Men with Asthenoteratozoospermia

Abstract

Male infertility is a worldwide problem, and many couples have suffered from it. Although cryopreservation is widely used for the long-term preservation of human sperm, sperm survival, and function post-thawing procedure may be strongly impaired and thus decrease the reproductive performance. This study examined whether adding Edaravone before cryopreservation could improve the post-thaw parameters (especially sperm motility) of cryopreserved spermatozoa in patients with asthenoteratozoospermia (AT). Semen samples were collected by masturbation from 25 men with AT and assessed following WHO standards. Samples were divided into three aliquots. The first aliquot remained untreated and freshly assessed (fresh group). The second aliquot was untreated, mixed with a cryopreservation medium, and cryopreserved (freeze group). The third aliquot was treated with ten µM Edaravone, mixed with cryopreservation medium, and cryopreserved (freeze + Edaravone group). The groups were assessed for motility, morphology, viability, apoptosis, lipid peroxidation, Mitochondrial Membrane Potential (MMP), DNA Fragmentation Index (DFI), glutathione (GSH), ATP, and ROS production. The freeze + Edaravone group significantly improved total sperm motility (P < 0.0001), progressive and non-progressive sperm motility (P < 0.01), viability (P < 0.0001), morphology (P < 0.001), DFI (P < 0.0001) and live sperm cells (P < 0.0001) post-thawing compared to freeze group. Additionally, the freeze + Edaravone group significantly decreased necrotic sperm cells (P < 0.0001), lipid peroxidation (P < 0.0001), and intracellular ROS production (P < 0.0001) post-thawing procedure. Furthermore, the freeze + Edaravone group significantly enhanced MMP jc-1 orange and green (P < 0.0001 and P < 0.01, respectively), ATP production (P < 0.0001), and GSH levels (P < 0.0001) post-thawing procedure compared to freeze group. Our findings proved that the freeze + Edaravone group of men with AT improved sperm survival and functions post-thawing procedure.

Keywords: Asthenoteratozoospermia; Cryopreservation; Edaravone; Sperm.