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 sperm 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, functional sperm concentration, motile sperm concentration), chromatin integrity, and DNA fragmentation were assessed before and after each processing method using SQA-V Gold sperm analyzer and 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.
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