Best washing method for oligoasthenospermic patients by evaluating DNA integrity by using comet assay study

Background: Sperm is more than just a mere transporter of paternal genetic information, although serving purposes other than conception. DNA integrity (DI) is regarded as a crucial aspect of semen quality and has great significance in predicting male fertility. Washing sperm is a form of preparation of sperm which is essential before processing intra-uterine-insemination since it removes chemicals from semen that might cause adversarial reactions in the uterus. During the process of washing, sperm gets separated from seminal fluid, which gradually improves the fertilizing capability of sperm, and numerous existing methods have faced complications with the determination of effective integrity in washing methods. The objective of the present study was to find the DI of the sperms through three washing methods that, include the density gradient (DG) method, swim up (SU) method, and magnetically activated cell sorting (MACS) method after the cryo-preservation (CP) for 3 months. Methodology: Semen samples were obtained from 40 oligoasthenospermic patients with progressive motility <32%, who underwent assisted reproductive technology procedure in Sumathi fertility center in Madurai between October 2021 and December 2021. Sperm washing is performed through three different mentioned techniques. The grades of the comet reflected five different classes from class 0 to class 4. These data were analyzed using one-way ANOVA test, and multiple comparisons were performed between these groups using Tukey's test. Results: The results showed a high level of normal sperm (68%) in the DG method than the SU (47%) and MACS (42%) sperm preparation techniques that specified by significant elevation (P < 0.001) of class 0 comet tails. Conclusion: It was observed that the DG method has the best and success rates for intracytoplasmic sperm injection and in vitro fertilization was relatively high.