The Application of Ultrasonic Vibration in Human Sperm Cryopreservation as a Novel Method for the Modification of Physicochemical Characteristics of Freezing Media

Abstract

The application of ultrasonic vibration was performed to modify the water molecules as the main compositions of the freezing medium used for human sperm cryopreservation. Different time periods of ultrasonic vibration (ULV) at the frequency of 28 kHz were applied for the evaluation of physicochemical properties of the water molecules. The most significant bubble size, zeta potential, and pH were obtained for the water molecules exposed to ultrasonic vibrations for 18 minutes and this time period was selected for further experiments due to the optimum results. In the next stage, semen samples were diluted with freezing medium containing ULV-exposed water and then cryopreserved. All the semen parameters were significantly reduced in cryopreserved groups as compared with the fresh control group. The highest percentage of total and progressive motility, viability, membrane and DNA integrity, and mitochondrial membrane potential were observed in frozen ULV compared with the frozen control. The rate of apoptosis in frozen ULV was significantly lower than that of in the frozen control. Furthermore, the gene expression ratios of α- and β-tubulins were significantly increased during cryopreservation, while the expression ratio of the tubulin polymerization promoting protein (TPPP) gene was decreased. Similar results were also observed when the protein levels of the genes mentioned earlier were evaluated by the ELISA method. Therefore, the changes in physicochemical properties of the freezing medium of human sperm cryopreservation using ULV can improve the quality of frozen products.

Unraveling Subcellular and Ultrastructural Changes During Vitrification of Human Spermatozoa: Effect of a Mitochondria-Targeted Antioxidant and a Permeable Cryoprotectant

“…Microtubules are composed of α-tubulin and β-tubulin. During traditional slow-freezing spermatozoa cryopreservation, high abundance of αand β-tubulins and low abundance of tubulin polymerization promoting proteins (TPPPs) have been reported (Desrosiers et al, 2006;Gholami et al, 2018;Dariush et al, 2019). Gholami et al (2018) reported more abundance of tubulin α-1B chain, tubulin α-3C/D chain, tubulin β-2C chain, and tubulin β-2B chain and low abundance of TPPP2 after human spermatozoa cryopreservation.…”

Electromagnetic field in human sperm cryopreservation improves fertilizing potential of thawed sperm through physicochemical modification of water molecules in freezing medium

“…Despite several advantages of this strategy, structural and biochemical damages caused by this process to sperm are the main challenge and drawback [4, 5] which lead to reduce the fertilizing potential of thawed sperm. This phenomenon is mostly related to sudden disruption of water molecule structures in cryopreservation media that can induce the production of ROS which, in turn, attacks to the sperm membrane and damage the sperm organelles resulting in sperm death [6, 7]. Water can form 16 different ice crystal structures during the freezing process; among these 16 structures, the hexagonal structure causes the greatest damage to cells during the cryopreservation [8].…”

Beneficial effects of trehalose and gentiobiose on human sperm cryopreservation

“…Despite sperm cryopreservation being widely used for fertility preservation, one should not ignore its detrimental effects such as motility malfunctions, reduction of viability rate, and other sperm parameter defects [ 22 , 28 ]. Usage of compounds with antioxidant activity such as sugars during cryopreservation indicated positive effects on sperm parameters [ 29 , 30 ].…”