Abstract

Background: The sperm DNA fragmentation index (DFI) is an important indicator reflecting the integrity of genetic material and may be affected by the cryopreservation–thawing process. Objective: To evaluate the effect of cryopreservation–thawing on DFI as well as the performance of an artificial intelligence (AI)-integrated automated sperm analysis system. Materials and Methods: A cross-sectional descriptive study was conducted on 30 male patients attending the Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy Hospital. Sperm DNA fragmentation before cryopreservation and after thawing was assessed using the sperm chromatin dispersion (SCD) method and analyzed by both the AI-integrated LensHooke X12 PRO system and manual microscopic evaluation. 48 TẠP CHÍ Y DƯỢC HUẾ - HUE JOURNAL OF MEDICINE AND PHARMACY ISSN 3030-4318; eISSN: 3030-4326 Results: The cryopreservation-thawing process showed a statistically significant increase in the DNA fragmentation index (DFI); however, the mean increase was low (3.15 ± 3.01%). After thawing, the proportion of spermatozoa with big halo significantly decreased, whereas the proportions of medium halo, small halo, no halo, and degraded spermatozoa significantly increased (p < 0.05). In samples with abnormal motility and morphology, DFI increased markedly (p < 0.001). Overall, the DFI results obtained from manual assessment and the AI-integrated automated analysis system demonstrated good agreement. Conclusion: Cryopreservation–thawing may increase sperm DNA fragmentation, particularly in lower- quality samples. The AI-integrated automated analysis system provided results comparable to those of manual evaluation while enabling the analysis of a larger number of spermatozoa and reducing observer- dependent bias, thereby supporting improved standardization of DFI assessment.