Sperm Preparation Techniques Affect Biological Parameters, Fertility Indices, and DNA Fragmentation in Patients with Oligo-Astheno-Teratozoospermia.

Abstract: In normozoospermia, the Albumin Gradient Filtration (AGF) and swim-up processing techniques are used to isolate spermatozoa with the highest biological parameters and specific sperm fertility indices (SSFI). In contrast, the highest number of sperms with the best motility rate, velocity, and SSFI, and the lowest DNA damage and nuclear fragmentation are isolated using the Density Gradient Centrifugation (DGC) preparation technique in Oligo-Astheno-Teratozoospermia (OAT) patients. The study aimed to evaluate the effect of DGC, AGF-1, AGF-2, and swim-up preparation techniques on biological parameters, including SSFI, chromatin integrity, and DNA fragmentation in patients with OAT and normozoospermia. One hundred men with normozoospermia and OAT participated in the study. Biological parameters (concentration, velocity, total motility, and normal morphology), SSFI [Progressively Motile Sperm Concentration-grade a (PMSCa), Progressively Motile Sperm Concentration-grade b (PMSCb), and Sperm Motility Index (SMI), Functional Sperm Concentration (FSC), Motile Sperm Concentration (MSC)], chromatin integrity, and DNA fragmentation were assessed before and after each processing methods using SQA-V Gold sperm analyzer, specific staining, TUNEL, and COMET assays. In OAT patients, the highest number of sperm with the best velocity, motility, and SSFI was demonstrated in the DGC method. However, the AGF-2 technique was the most efficient method in normozoospermic patients. Sperm DNA damage significantly decreased following preparation with DGC, AGF-2, and swim-up techniques. Overall, the findings highlight that DGC is the most beneficial method for OAT patients, whereas AGF-2 and swim-up techniques are more suitable for normozoospermic individuals. These comparisons emphasize the need to tailor techniques according to patient-specific sperm profiles.

Lay summary: This study investigated how different laboratory methods for preparing sperm would help improve fertility treatment outcomes for men with fertility problems. We assessed four commonly used techniques to see how well they could choose the healthiest sperm from 100 men, both with and without fertility issues. We aimed to improve sperm quality by selecting those that move faster, have fewer abnormalities, and contain less damaged DNA. We revealed that men with poor sperm quality benefited most from a method that separates sperm based on how heavy they are. In contrast, men with normal sperm quality responded better to a different technique that uses a protein-based filter. All the methods tested were helpful in reducing sperm DNA damage, but two of them were more effective in preserving sperm health based on men's conditions. Our findings suggest that using the right sperm preparation method for each individual can increase the chances of success in fertility treatments.