RESUMEN
Isoflurane, an inhalational anesthetic from the halogenated group, has been increasingly used in the medical and scientific fields. Due to its characteristics, it is capable of inducing anesthesia quickly and quietly; however, the adverse effects resulting from its use have not yet been fully elucidated, especially with regard to reproductive aspects. Considering its common use in research laboratories, whether for performing surgical procedures or for prior exposure to euthanasia, knowledge about its interference in sperm parameters of experimental models characterizes an important study goal. The aim of the present study was to determine the interference of acute exposure to isoflurane on the sperm quality of mice, both immediately previous to euthanasia and in later evaluation, twenty days after a single anesthetic exposure. Our results demonstrate that acute anesthetic exposure reduces sperm motility and is responsible for the formation of damaged sperm cells that are prone to apoptosis, which may affect the outcome of reproductive experiments even 20 days after exposure.
RESUMEN
Around the world, many couples have turned to in vitro fertilization as a viable solution to fertility issues. Low-density lipoprotein (LDL) is a protein best known for transporting fat molecules throughout the body, but it has also been shown to protect sperm cells during cryopreservation due to its micellar structure. In the present study, we aimed to evaluate different protocols for the preparation of nanoparticles from egg yolk plasma (EYP) containing LDL to improve the viability of cryopreserved canine semen. EYP was subjected to three distinct treatments: ultrasonification in an ultrasound bath at 40 kHz for 30 min (LDL-B); ultrasonification via an ultrasound probe at 50% amplitude for 30 min (LDL-P); and high-pressure homogenization at 10,000 PSI for six cycles (LDL-H). Sperm quality was assessed after thawing using computer-assisted sperm analysis and flow cytometry. The results revealed that compared to the EYP control, the LDL-P formulation presented significantly higher efficiency (p < 0.05) in maintaining total and progressive sperm motility, sperm membrane integrity and fluidity, and levels of intracellular reactive oxygen species. The LDL-P nanoparticles had an average size of approximately 250 nm, a PDI value of 0.3, and -1.15 mV of zeta potential, which are very important because it is an indicator of the stability of a colloidal dispersion. Therefore, we conclude that ultrasonication of EYP using a probe is an efficient method for the preparation of LDL nanoparticles that would enhance the cryoprotection of semen during freezing.