Tehran University of Medical Sciences Journal of Family and Reproductive Health 1735-8949 19 4 2025 12 31 301 309 Mehdi Mokaddem Intelligent Automation and BioMed Genomics Laboratory, Faculty of Sciences and Techniques of Tangier, Abdelmalek Essaadi University, Tetouan, Morocco Zeineb Zian Intelligent Automation and BioMed Genomics Laboratory, Faculty of Sciences and Techniques of Tangier, Abdelmalek Essaadi University, Tetouan, Morocco Amr Kchikich Laboratory of Biotechnological Valorization of Microorganisms, Genomics, and Bioinformatics, Faculty of Sciences and Techniques, University Abdelmalek Essaadi, Tetouan, Morocco Noureddine Louanjli Laboratory of Medical Analysis, Andrology Department, LABOMAC, Casablanca, Morocco Naima Nourouti Intelligent Automation and BioMed Genomics Laboratory, Faculty of Sciences and Techniques of Tangier, Abdelmalek Essaadi University, Tetouan, Morocco Amina Barakat Intelligent Automation and BioMed Genomics Laboratory, Faculty of Sciences and Techniques of Tangier, Abdelmalek Essaadi University, Tetouan, Morocco Moudou Mbaye Laboratory of Medical Analysis, Andrology Department, LABOMAC, Casablanca, Morocco Mohcine Mechita Intelligent Automation and BioMed Genomics Laboratory, Faculty of Sciences and Techniques of Tangier, Abdelmalek Essaadi University, Tetouan, Morocco 2025 10 26 2025 12 30 Objective: This study aimed to evaluate the effects of hyaluronic acid (HA) supplementation on motility, vitality, and DNA integrity in Moroccan fresh and cryopreserved sperm samples. Materials and methods: A cohort of 60 male participants was divided into three groups, including normozoospermic, asthenozoospermic, and cryopreserved normozoospermic samples. Advanced microscopy and computer-assisted semen analysis (CASA) were used to analyze the sperm parameters before and after HA supplementation. DNA fragmentation was analyzed using TUNEL assay, while chromatin condensation was evaluated by aniline blue staining. Statistical analysis was performed using ANOVA and Duncan's test. Results: HA supplementation improved sperm motility and vitality (p < 0.05) in the normozoospermic group without altering DNA fragmentation or chromatin condensation. In the asthenozoospermic group, motility and vitality were significantly improved (p < 0.05). In the cryopreserved normozoospermic group, HA improved motility and vitality after thawing (p < 0.05). Conclusion: HA is a promising adjuvant for enhancing the quality of both fresh and cryopreserved sperm for ART without compromising DNA integrity. Studies need to be extended for further exploration of the mechanism of action and optimization of clinical protocols toward its use in routine practice, particularly for diverse populations and in the more difficult cases of infertility.   https://jfrh.tums.ac.ir/index.php/jfrh/article/view/3543 https://jfrh.tums.ac.ir/index.php/jfrh/article/download/3543/744