In vitro application of Ceratonia siliqua improved sperm parameters and chromatin quality after vitrifacation in normozoospermic aged men

Abstract

Background: Vitrification is the main technique in the assisted reproductive technique (ART) labs. Sperm vitrification exposes sperm to damage. The aim of the present study was to evaluate the in vitro effect of Ceratonia siliqua (C. siliqua) application on sperm parameters and chromatin quality in normozoospermic aged men. Semen samples (n = 40) were collected from normozoospermic men over 45 years old. Each specimen was divided into four aliquots to form the subsequent groups: fresh (group I), vitrification without treatment (group II), vitrification with the medium supplemented by 20 μg/ml C. siliqua (group III), and vitrification with the thawing medium supplemented by 20 μg/ml C. siliqua (group IV). Sperm progressive motility, normal morphology and viability were assessed. Also, sperm chromatin quality was evaluated by aniline blue (AB), toluidine blue (TB), and sperm chromatin dispersion (SCD) staining. Results: Vitrification caused a significant decrease in sperm progressive motility, normal morphology and viability as well as chromatin quality compared to fresh samples (p < 0.05). Supplementation of vitrification/thawing medium with C. siliqua significantly improved sperm progressive motility, normal morphology, viability, and chromatin quality compared to vitrification without any supplementation (p < 0.05). Conclusions: The study showed that C. siliqua can improve the detrimental effect of vitrification on sperm parameters and chromatin quality of normozoospermic aged men.