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Protective Effects of Sesamol-Loaded Nanostructured Lipid Carriers on Human Spermatozoa During the Cryopreservation
Vahid Shokri-Asl1, Hamed Hajipour1, Maryam Pashaiasl1
1Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Introduction:
Sperm cryopreservation induces intracellular oxidative stress. Sesamol (SE), as an antioxidant, mitigates cryodamage caused by spermatozoa cryopreservation. This study was designed to evaluate the protective effects of SE-loaded nanostructured lipid carriers (SE-loaded NLCs) on frozen sperm quality.
Methods:
SE-loaded NLCs were synthesized using hot homogenization, followed by characterization for particle size, morphology, and encapsulation efficiency (EE). Normozoospermic semen samples were collected from 20 men and prepared according to the WHO 2021 guidelines. The samples were allocated into experimental groups of fresh, frozen-thawed control, SE-treated, blank NLCs, and SE-loaded NLCs. Following semen cryopreservation and thawing protocols, sperm quality was evaluated regarding motility, vitality, morphology, membrane and acrosome integrity, mitochondrial membrane potential (MMP), and DNA fragmentation using established staining and microscopy techniques. Statistical analysis was also performed at the significance level of 0.05.
Results:
NLCs were successfully prepared with a mean particle size of 154 nm, zeta potential of -17.6 mV, and EE of 72 ± 2.64%. Fluorescence microscopy confirmed the efficient uptake of NLCs by spermatozoa within 60 minutes. The optimal SE dose was determined as 50 µM for cryoprotection. Sperm treated with SE-loaded NLCs showed improvements (p < 0.05) in motility, viability, membrane and acrosome integrity, MMP, and reduced (p < 0.05) DNA fragmentation than other groups.
Conclusion:
Application of SE-loaded NLCs showed accelerated protective effects compared with free SE, highlighting the associated potential in the enhancement of sperm quality during cryopreservation. SE-loaded NLCs effectively enhanced sperm quality indices during cryopreservation. Thus, SE-loaded NLCs represent a promising strategy to mitigate cryoinjury and improve fertility preservation outcomes.
