The effect of sperm cryoprezervation in different semen concentrations on sperm vivity and DNA fragmentation

In about 20% of infertile couples, the male factor is the main cause. If the combination of male and female factors is included, this rate is reached to 30% -40%. Nowadays, routine semen analysis is used to evaluate male infertility, but in 15% of infertile men, despite the results of normal semen analysis, the definitive diagnosis of infertility cannot be established by routine semen analysis. Therefore, the need for new markers to differentiate between fertile and infertile men and predict the outcome of pregnancy has increased and attention has been focused on sperm DNA integrity. It has been proven that sperm DNA damage is more common in infertile men than fertile men and that sperm DNA damage negatively affects fertility potential in these patients. Impaired semen parameters such as low number, motility, and abnormal morphology are often associated with high sperm DNA damage, but in patients with normal semen parameters In addition, there are concerns about the potential consequences of the use of sperm with damaged DNA in intrastoplasmic sperm injection (ICSI). There are different levels of damage to DNA due to various internal and external causes. The main damages in DNA are; degradation of chromatin structure, oxidation of DNA bases, mismatch and suppression of tubulin polymerization, chemical change of bases, anomalies in chromatin structure, breakage of DNA chain, DNA-DNA and DNA-protein crossings, DNA and DNA mutations. Sperm cryopreservation is a frequently used method in assisted reproductive therapy. After thawing, it is the target that sperms are damaged at least. In our study; The aim of this study is to investigate whether there is a change in sperm DNA condensation after sperm freezing thawing process, to determine whether this change is correlated with decreasing motility, and also to investigate whether semen raw state or post-wash ice cream has an effect on motility, morphology and DNA condensation. In our study, 33 male patients aged between 18-50 years who applied to Biruni University Hospital Urology Outpatient Clinic were planned to work with waste sample after routine semen analysis. In order to investigate the effect of sperm cryopreservation at different semen concentrations on sperm viability and sperm DNA fragmentation, first semen analysis was performed according to World Health Organization (WHO) criteria for all patients to be included in the study and sperm concentration, motility and morphology were evaluated. For the determination of sperm viability, eosin staining was performed and evaluated under light microscope. In order to investigate the effect on sperm DNA fragmentation, smear was prepared by fixing 1/3 acetic acid in methanol mixture for 30 minutes at room temperature or -40 degrees (refrigerated) and stained with acridine ratio and supported by Biruni University with immunofluorescence microscope.