Niosomal nanocarriers loaded with artemisinin has potential to enhance the functional characteristics of cryo-preserved equine spermatozoa.

Cryopreservation is a crucial method for the preservation of genetic materials. Nevertheless, this technique can have adverse effects due to cryo-oxidative damage. The primary objective of this study was to investigate the cryo-protective effects of niosomal nanocarriers loaded with artemisinin (NN-Art) on the functional attributes of equine spermatozoa. The parameters examined included sperm motility and kinematics through computer-assisted sperm analysis (CASA), plasma membrane integrity via the hypoosmotic swelling test, viability using the eosin-nigrosin staining technique, morphology with Hancock's solution, and lipid peroxidation through the thiobarbituric acid reactive substances assay. Additionally, flow cytometry was applied to assess mitochondrial membrane potential, reactive oxygen species (ROS) levels, sperm chromatin structure, and apoptotic-like changes. The CASA indicated a greater enhancement in the total and progressive motility and as well straight line velocity following the addition of NN-Art into the cryo-diluent medium compared with the conventional form of artemisinin and control (P < 0.05). Moreover, the incorporation of cryo-diluent medium with NN-Art demonstrated a greater efficiency in the improvement of viability, proportion of spermatozoa abnormality, plasma membrane functionality, and lipid peroxidation (P < 0.05). The flow cytometry assessment demonstrated a significant enhancement in mitochondrial potential activity, a reduction in the level of ROS, and an increase in the proportion of live spermatozoa along with a decrease in late apoptotic stallion post-thawed spermatozoa treated with both Art and specifically NN-Art. The results of this study suggest that NN-Art exhibit enhanced antioxidant capabilities, which are crucial for improving the functional quality of equine spermatozoa by mitigating cryo-oxidative stress.