Protective effect of crocetin on bovine spermatozoa against oxidative stress during in vitro fertilization

Abstract

Oxidative stress is one of the major factors that contribute to poor semen quality and low rates of in vitro fertilization. Crocetin, a main constituent of saffron (Crocus sativus L.) possesses potent antioxidant activity, by scavenging reactive oxygen species (ROS) and/or enhancing the activity of intracellular antioxidant enzymes. The aim of this study was to investigate, for the first time, the effect of crocetin on the quality characteristics of bull spermatozoa and fertilization rate. For this reason, frozen/thawed bovine spermatozoa were incubated with crocetin (1, 2.5, and 5 μm), for 120 or 240 min, in the presence of a negative control, and evaluated in terms of motility, viability, acrosomal status, DNA fragmentation index, intracellular ROS, and lipid peroxidation. In order to evaluate the impact of crocetin on cleavage and blastocyst rate, the compound was added in the IVF medium at the previously identified optimal concentration (2.5 μm). The results indicate that incubation of spermatozoa with 2.5 μm of crocetin resulted in a statistically significant lower production of superoxide anion and hydrogen peroxide, lower lipid peroxidation, and in better maintenance of motility parameters, viability, and acrosomal integrity, with a very small number of cells with DNA fragmentation, compared to the other groups (p < 0.05). The presence of crocetin (2.5 μm) in the fertilization medium also resulted in a significant increase in acrosome-reacted spermatozoa and blastocyst production, compared to the control group (p < 0.01). These data indicate that crocetin (2.5 μm) positively affects bovine sperm quality characteristics during a 240-min incubation and improves its fertilizing ability, directly and/or indirectly, by regulating ROS concentration and lipid peroxidation.

Antioxidant effects of the essential oil ofSyzygium aromaticumon bovine epididymal spermatozoa

“…Sperm concentration was determined using a Neubauer counting chamber, and the study was only conducted when sperm cells had more than 60% motility. Finally, the sperm suspension was centrifuged (300 g for 10 min) and suspended in sperm Tyrode's albumin lactate pyruvate (sperm‐TALP) medium (100 mmol/L NaCl, 3.1 mmol/L KCl, 25 mmol/L NaHCO 3 , 0.29 mmol/L NaH 2 PO 4 , 21.6 mmol/L Na lactate, 2 mmol/L CaCl 2 , 1.5 mmol/L MgCl 2 , 10 mmol/L Hepes, 0.6% bovine serum, 1 mmol/L sodium pyruvate and 1% antibiotic–antimycotic solution in water for embryo transfer, according to Sapanidou et al, ).…”

“…However, for in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI), spermatozoa are exposed to external factors, such as oxygen tension, light interference, heat interference and medium constituents, which may accentuate ROS production and cause oxidative stress (Agarwal, Said, Bedaiwy, Banerjee, & Alvarez, ). Therefore, the addition of antioxidants in vitro may be effective for maintaining sperm viability to achieve greater fertilisation success (Sapanidou et al, ).…”

“…Several natural substances have been studied for antioxidant treatment of bovine spermatozoa (Santos, Borges, Queiroz Neta, Bertini, & Pereira, ), both as isolated substances (Sapanidou et al, ; Tvrdá, Kováčik, Tušimová, Massányi, & Lukáč, ; Tvrdá et al, ) and as essential oils and extracts (Sapanidou et al, ). Despite the positive results already reported, there is still no specific antioxidant for spermatozoa being used in assisted reproduction techniques and the results still need to be improved (Santos et al, ).…”

Anethole improves blastocysts rates together with antioxidant capacity when added during bovine embryo culture rather than in the in vitro maturation medium

“…Oxidative stress during IVM (Combelles et al, 2009;Sovernigo et al, 2017), IVF (Sapanidou et al, 2016), and IVP (Simões et al, 2013;Sá et al, 2019), is one of the factors limiting IVP success. Therefore, antioxidants are key compounds to improve in vitro embryo production (Torres et al, 2018(Torres et al, , 2019.…”

Cellular Basis and Optimization of Cryopreservation Survival of Bull Sperm = Base celular y optimización de la supervivencia de la criopreservación de esperma de toro

“…Crocin is a glucosyl ester of crocetin, a water-soluble carotenoid extracted from the perennial herb saffron (Crocus sativus). These carotenoids act as antioxidants, with a high ability to scavenge the free radicals (Bors et al, 1982;Rios et al, 1996;Bakhtiary et al, 2014;Sapanidou et al, 2016;Esposito et al, 2019), especially superoxide anion (Galano et al, 2010).…”