This study developed and characterized Ziziphus spina-christi leaf extract (ZSCLE)-loaded hyaluronic acid liposomes (ZSCLE-HS) and assessed their impact on improving assisted reproductive technology such as semen cryopreservation. The ZSCLE was prepared, and the active ingredients were identified and quantified using HPLC. The optimized ZSCLE-HS was assessed for entrapment efficiency (EE, %), particle size, TEM, FTIR, XRD analysis, and stability over a 4-month period at 4 °C. The ex vivo protective function of the optimized ZSCLE-HS on cryopreserved ram sperm was evaluated. The optimized ZSCLE-HS was spherical in shape with an average particle size of 426.16 ± 31.79 nm, a zeta potential of -41.03 ± 0.4 mV, a polydispersity index of 0.20 ± 0.1, and % EE of 87.08 ± 2.82. Ex-vivo trials demonstrated that ZSCLE-HS (50-100 μg) supplementation in freezing extenders significantly enhanced cryopreserved ram sperm quality by improving sperm function, kinematics, and antioxidant capacity, while mitigating apoptosis, lipid peroxidation, and acrosome damage, and preserving sperm ultrastructure. Molecular docking assays predicted strong binding affinities between naringenin and the sperm proteins GPx and NADH. Our ex-vivo findings suggest that ZSCLE-HS has the potential to alleviate oxidative stress, enhance ram sperm quality following cryopreservation, and ultimately improve the success rates of assisted reproductive technologies.