Background: Suboptimal human semen handling in vitro may induce sperm damage.However, the effects of semen swim-up, pellet swim-up, density gradient, and density gradient followed by SU on sperm motility, morphology, DNA fragmentation, acrosome reaction, intracellular reactive oxygen species, and mitochondrial activity were not fully understood.Objectives: To study the impact of four sperm preparation techniques on sperm functional parameters.
Materials and methods:This study was conducted on 60 infertile men with a minimum sperm concentration of 20 × 10 6 /ml and total sperm motility of ≥30%. Each raw semen sample was divided into four aliquots. Each aliquot was prepared by one of the tested techniques. Various sperm characteristics were assessed before and after sperm preparation.Results: Density gradient and density gradient followed by SU resulted in significantly higher DNA fragmentation percentages compared with semen swim-up (p < 0.001 and p < 0.001, respectively) and pellet swim-up (p < 0.001 and p < 0.001, respectively). Significantly higher percentages of spermatozoa with intact acrosome were detected in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. The percentage of reactive oxygen species-positive spermatozoa was significantly higher after pellet swim-up (p < 0.001), density gradient (p < 0.001), and density gradient followed by SU (p < 0.001) than raw semen. In addition, the percentages of 100% stained midpiece (active mitochondria) were significantly higher in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen.
Discussion and conclusion:To the best of our knowledge, this is the first report comparing the impact of these techniques on various sperm functional parameters.Semen swim-up was more effective than density gradient in selecting better spermatozoa in terms of DNA integrity, reactive oxygen species levels, acrosome status, and
<b><i>Introduction:</i></b> A firm consensus on the effectiveness of psychological interventions during infertility treatment has not been reached yet in terms of mental health and pregnancy rates. Moreover, the influence of these interventions on embryo cleavage kinetics has not been investigated. <b><i>Objective:</i></b> The aim of this work was to study whether stress management in couples undergoing an intracytoplasmic sperm injection (ICSI) cycle influences stress levels, mitochondrial DNA (mtDNA) levels in granulosa cells, and cleavage-stage embryos. <b><i>Methods:</i></b> Infertile couples were randomized into a treatment as usual (TAU) group (<i>n</i> = 30) and stress management program (SMP) group (<i>n</i> = 29) at the beginning of an ICSI cycle. Couples in the SMP group attended education and relaxation sessions at each visit to the clinic for folliculometry. The perceived stress scale (PSS) was used to assess stress levels at the beginning and end of the cycle. Moreover, mtDNA levels of granulosa cells and embryo morphokinetics were evaluated. <b><i>Results:</i></b> Post-intervention, women in the SMP group had significantly lower PSS scores than their initial PSS (<i>p</i> < 0.001; effect size, ES = 0.5) and than the final PSS of the TAU group (<i>p</i> = 0.02; ES = 0.09). Additionally, mtDNA levels were significantly lower in luteal granulosa cells of the SMP group than the TAU group (<i>p</i> = 0.02). An earlier time of pronuclei appearance (<i>p</i> = 0.03) and time to 2 cells (<i>p</i> = 0.015) and a faster time to full compaction (<i>p</i> = 0.045) were detected in the embryos of the SMP group compared with the TAU group. <b><i>Conclusion(s):</i></b> The implemented program may reduce stress levels, retard first embryo cleavage, and accelerate embryo compaction. Further studies with an active control group are needed to confirm these results.
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