Methods Objectives Results Results Conclusions References Viability and DNA fragmentation improvement by adding [6]–Gingerol to freezing medium Shadi Zekri¹٭, Dr. Marjan Sabbaghiyan², Dr. Abdolhossein Shahverdi³ 1. Biology Department, Research and Science Branch of Islamic Azad University, Tehran, Iran Objectives Results Results The function of [6] – Gingerol as an Antioxidant resulted in DNA integrity and improved sperm viability. Evaluation of sperm DNA fragmentation Means (±SEM) with different superscripts differ (a, b, c) P<0.05 Green fluorescence represented sperm with normal double stranded DNA. Red fluorescence represented sperm with single stranded DNA. At present, cryopreservation of sperm is a valuable method for maintaining reproductive performance in men with abnormal sperm such as azoospermia, oligospermia or men with cancer. Despite the successes achieved in this method, during this process, sperm injuries happen that reduce the percentage of sperm viability [1]. Sperm freezing causes to enhance the production of free radicals that can easily penetrate the membranes and per oxidative damages to DNA, proteins and lipids in the cell, and also increases the level of peroxidation of lipids. This decreases membrane fluidity, causes the loss of sperm function and fragmentation of DNA. Oxidative stress induced by freezing and thawing can induce apoptosis and impairment in the mitochondrial membrane potential [2,3]. Antioxidants are factors that reduce the oxidative stress by breaking down the oxidative chain reactions. The collaboration of these agents is an effective way to improve the vitally and function of sperm during the process of freezing and thawing [4]. Our aim was to investigate the effects of [6] -Gingerol as an antioxidant during the sperm freezing and thawing process. groups: fresh, freezing and freezing with [6] –Gingerol. Our results revealed that freezing with [6]–Gingerol increase sperm viability in comparison with control group (64.05 ± 1.32 and 56.45 ± 1.26). Sperm freezing leads to an increasing of free radicals,[5] thus adding [6]-Gingerol as an antioxidant reduced the level of free radicals and increased the viability of sperm after freezing. Evaluation of sperm viability Means (±SEM) with different superscripts differ (a, b, c) P<0.05 Sperm viability assessment : Eosin-Nigrosin Test A: Live sperm, B: Dead sperm Whereas this antioxidant had little effect on motility and progressive motility in freezing with [6] – Gingerol group. Finally DNA fragmentation was higher in treated group (37.89 ± 1.30) as compared to control group (42.9 ± 0.95). DNA fragmentation during cryopreservation has been linked to the production of ROS during the freezing and thawing process [6]. A Methods B Conclusions Samples were collected from 20 normospermic men referred to the Infertility Institute & Research Center and classified into two groups: freezing (control) group and freezing with a medium supplemented with 10 μM of [6] –Gingerol (treated). Then the sample were frozen with fast technique and thawing. Then the Sperm motility, progressive motility and viability were measured based on WHO criteria with computer- assisted-semen analysis (CASA), Eosin-Nigrosin staining. Also, DNA fragmentation were determined with Acridin orange test (AOT). The mentioned parameters were evaluated in 3 We concluded that adding of the [6] -Gingerol as an antioxidant to the freezing and thawing medium can improve the sperm cryopreservation condition. References Oberoi B, Kumar S, Talwar P. Study of human sperm motility post cryopreservation. Medical journal armed forces India. 2014; 70(4):349-53. Bansal AK, Bilaspuri G. Impacts of oxidative stress and antioxidants on semen functions. Veterinary Medicine International. 2011:1-7. Naeini ZK, Bafrani HH, Nikzad H. Evaluation of ebselen supplementation on cryopreservation medium in human semen. Iranian journal of reproductive medicine. 2014; 12(4):249. Nadjarzadeh A, Sadeghi M, Amirjannati N, Vafa M, Motevalian S, Gohari M, et al. Coenzyme Q10 improves seminal oxidative defense but does not affect on semen parameters in idiopathic oligoasthenoteratozoospermia: a randomized double-blind, placebo controlled trial. Journal of endocrinological investigation. 2011; 34(8):e224-e8. Di Santo M, Tarozzi N, and Nadalini M, Borini A. Human sperm cryopreservation: update on techniques, effect on DNA integrity, and implications for ART. Advances in urology. 2012: 1-12. Khodayari Naeini Z, Hassani Bafrani H, Nikzad H. Evaluation of ebselen supplementation on cryopreservation medium in human semen. Iran J Reprod Med 2014;12( 4.) : 249-256 .