Abstract

Purpose: The quantitatively measured sperm DNA damage and disturbed chromatin condensation indexes (DNA fragmentation index DFI% and %high DNA staining index %HDS) and their relationships with sperm quality in normospermic men with unexplained infertility were investigated. The aim was also highlighting the impact of age on both DFI and CMA3 staining and on sperm quality, and their associations with male infertility. Methods: In this retrospective study, conventional semen tests, including sperm motility and morphological evaluations and DFI and disturbed chromatin condensation indexes (DFI, CMA3) were performed according to the World Health Organization (WHO) 2021 criteria. DFI and CMA3 were evaluated using sperm chromatin dispersion (SCD) and chromomycin A3 (CMA3) staining assays and then correlation and regression analysis were done. Results: By analyzing SCD and CMA3 results, notable differences were found in sperm parameters among different DFI and CMA3 groups (all p < 0.05). It was found that in the male fertility quality sperm concentration, progressive (PR)/non-progressive (NP) motility, immobility, and morphology were significantly associated with sperm DFI and CMA3, but not with age (p < 0.05). Conclusions: Sperm molecular index DFI and CMA3 negatively affect male fertility quality through semen parameters (sperm concentration, motility, and morphology). SCD and CMA3 indexes show a significant negative correlation with sperm quality in normospermic males, which highlights its role in the assessment of male fertility potential and molecular evaluation of infertility treatment. © 2025 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.