RESUMEN
This study aims to improve the freezing-thawing process of human sperm using a static magnetic field. The study included 25 normozoospermic human samples. After an initial evaluation of sperm parameters, samples were prepared by the direct swim-up method. Before freezing, sperm motility, viability, morphology, acrosome reaction and DNA fragmentation rate were assessed. The samples were divided into 4 groups: 0, 1, 5 and 10 mT, and each group was frozen by the rapid freezing method. After thawing, the parameters were re-evaluated and compared between groups. Sperm motility decreased significantly during cryopreservation in all groups. The static magnetic field did not protect against decreased progressive motility after freezing, but the total sperm motility was significantly higher in the 10 mT group compared to the other groups. Sperm viability was higher in the 10 mT group than in the other groups. There was no significant difference in the rate of normal sperm morphology after freezing. The rate of spermatozoa with intact acrosome decreased after freeze-thawing, and the static magnetic field did not protect against the acrosome reaction. The rate of DNA integrity was significantly higher in the 10 mT group compared to the other groups. A static magnetic field with an intensity of 10 mT improved sperm viability and DNA integrity compared to other groups. However, it did not provide significant protection against decreased sperm motility or acrosome reaction.
Asunto(s)
Reacción Acrosómica , Supervivencia Celular , Criopreservación , Fragmentación del ADN , Campos Magnéticos , Preservación de Semen , Motilidad Espermática , Espermatozoides , Humanos , Masculino , Criopreservación/métodos , Espermatozoides/fisiología , Preservación de Semen/métodos , Congelación , AdultoRESUMEN
The role of cytoplasmic fragmentation in human embryo development and reproductive potential is widely recognized, albeit without standard definition nor agreed upon implication. While fragmentation is best understood to be a natural process across species, the origin of fragmentation remains incompletely understood and likely multifactorial. Several factors including embryo culture condition, gamete quality, aneuploidy, and abnormal cytokinesis seem to have important role in the etiology of cytoplasmic fragmentation. Fragmentation reduces the volume of cytoplasm and depletes embryo of essential organelles and regulatory proteins, compromising the developmental potential of the embryo. While it has been shown that degree of fragmentation and embryo implantation potential are inversely proportional, the degree, pattern, and distribution of fragmentation as it relates to pregnancy outcome is debated in the literature. This review highlights some of the challenges in analysis of fragmentation, while revealing trends in our evolving knowledge of how fragmentation may relate to functional development of the human embryos, implantation, and pregnancy outcome.
Asunto(s)
Citoplasma , Desarrollo Embrionario , Resultado del Embarazo , Humanos , Femenino , Embarazo , Desarrollo Embrionario/fisiología , Citoplasma/metabolismo , Citoplasma/fisiología , Implantación del Embrión/fisiologíaRESUMEN
Background: Due to myelin and axonal insults in multiple sclerosis individuals, motor coordination problems and endocrine imbalance may develop. Objective: This study aims to evaluate the role of chronic demyelination on the hypothalamic-pituitary-gonadal axis in the mouse model of multiple sclerosis. Materials and Methods: 20 adult C57/BL6 male mice were divided into 2 groups (n = 10/each) as follows: the control group (CONT) received a regular diet for 17 wk; and the experimental group (cuprizone [CPZ]) was fed with 0.2% CPZ for 12 wk and, then CPZ was withdrawn for 5 wk. Serum testosterone, histopathology of the brain and testis, and sperm analysis were evaluated. Results: The hypothalamic myelin content was significantly decreased in the arcuate nucleus following the 12 wk of CPZ consumption compared to the CONT group, and the statistical difference remained until 17 wk. Testosterone levels declined significantly in the CPZ group compared to the CONT group in the 12 th and 17 th wk. A significant decrease was observed in the height of the seminiferous epithelium and the interstitial tissue area, and the number of seminiferous epithelial cells in the CPZ group compared to the CONT group in the 12 th and 17 th wk. The sperm count, motility, and viability in the CPZ group significantly decreased compared to the CONT group in the 12 th and 17 th wk of the study. Conclusion: Chronic demyelination induced by CPZ intoxication, maybe through damage to the hypothalamus arcuate nucleus, leads to the hypothalamic-pituitary-gonadal axis disturbance and damage to the testis and spermatogenesis subsequently.
RESUMEN
The cryopreservation procedure decreases sperm quality, causing certain changes at structural and molecular levels affecting fertilizing ability. We aimed to investigate the impacts of human adipose-derived mesenchymal stem cells (HAd-MSCs) conditioned medium (CM) on the protection of human sperm from cryoinjury. Thirty normal semen specimens were evaluated in this study. Each specimen was separated into six groups and enhanced with varying concentrations of human Ad-MSCs-CM (0, 10, 30, 50, 70, and 100%). Sperm motility, viability, morphology, apoptosis, mitochondrial potential, and lipid peroxidation, and DNA fragmentation were evaluated before freezing and after thawing. The results showed that the total motility was preserved in 10% human Ad-MSCs-CM group. Also, DNA fragmentation was significantly lower in 10% compared to 0% human Ad-MSCs-CM (63.62 ± 17.72% vs.76.46 ± 4.87%, respectively, P < 0.004). Human Ad-MSCs-CM in groups of 10, 30, 50, and 70% reduced lipid peroxidation. The normal sperm morphology rate, mitochondrial membrane potential, and apoptosis showed no significant differences across various groups. It seems that human Ad-MSCs-CM can protect the sperm parameters during the cryopreservation by decreasing cryoinjury.
Asunto(s)
Criopreservación , Células Madre Mesenquimatosas , Preservación de Semen , Motilidad Espermática , Espermatozoides , Humanos , Criopreservación/métodos , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Medios de Cultivo Condicionados/metabolismo , Medios de Cultivo Condicionados/farmacología , Espermatozoides/efectos de los fármacos , Espermatozoides/metabolismo , Motilidad Espermática/efectos de los fármacos , Preservación de Semen/métodos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Tejido Adiposo/citología , Tejido Adiposo/metabolismo , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Fragmentación del ADN/efectos de los fármacos , Apoptosis/efectos de los fármacos , Análisis de Semen , AdultoRESUMEN
Human sperm cryopreservation is a routine procedure in assisted reproductive technology, but it has detrimental effects on different sperm parameters due to oxidative stress. Our objective was to assess the impacts of hydroxytyrosol (HT), as an antioxidant, on human sperm parameters following cryopreservation. In the first phase, 20 normal human semen samples were cryopreserved using the rapid freezing method with different concentrations of HT including 0, 50, 100, 150, and 200 µg/mL. In the second phase, 20 normal semen samples were collected and cryopreserved with 50 and 100 µg/mL HT. The beneficial effects of HT were determined by evaluation of motility (computer-assisted sperm analysis; CASA), viability (Eosin-nigrosine stain), DNA integrity (sperm chromatic dispersion test, SCD), reactive oxygen species (DCF and DHE staining by flowcytometry) lipid peroxidation (malondialdehyde, MDA test) and mitochondrial membrane potential (JC1 staining by flowcytometry) of sperm after cryopreservation. After thawing, sperm motility had an increasing trend in 50 and 100 µg/mL HT groups in comparison with other groups, althought the difference was not significant. However, sperm viability was significantly increased at 50 and 100 µg/mL HT. Our data also showed that sperm DNA fragmentation was significantly decreased after thawing at 100 µg/mL in comparison with 0 and 50 µg/mL HT. However, the level of intracellular reactive oxygen species, lipid peroxidation and mitochondrial membrane potential were not significantly different between groups. Our results showed that HT may have protective effects on the viability and DNA integrity of human sperm during the freezing-thawing process.