The importance of incorporating sperm DNA fragmentation testing in male infertility diagnostic routine

Garcia-Segura, Sergio

doi:10.21037/tau-22-572

Cited by 1 publication

(1 citation statement)

References 19 publications

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“…DNA strand breaks were detected via alkaline comet assay (SSB, DSB), TUNEL assay (SSB, DSB), and γ-H2AX-53BP1 co-localization focus assay (DSB). Cell death was quantified by caspase-3/7 apoptosis assay (21,24,25). In the late phase of apoptosis, fragmentation of genomic DNA occurs, which also leads to the formation of DNA DSBs and thus to positive comet and TUNEL signals, while such cells display a pan-γ-H2AX formation in their nuclei (18,26,27).…”

Section: Discussionmentioning

confidence: 99%

Short-term Effects of Non-invasive Physical Plasma Treatment on Genomic Stability

GELBRICH,

MUHTADI,

SCHERTHAN

et al. 2023

In Vivo

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Background/Aim: The application of non-invasive physical plasma (NIPP) generates reactive oxygen species. These can lead to chemical oxidation of cellular molecules including DNA. On the other hand, NIPP can induce therapeutically intended apoptosis, which also leads to DNA fragmentation in the late phase. Therefore, to assess unwanted genotoxic effects, the formation of DNA damage was investigated in this study in discrimination from apoptotic processes. Materials and Methods: Mutation events after NIPP application were analyzed in CCL-93 fibroblast cells using the hypoxanthine phosphoribosyl transferase assay. Additionally, DNA single-strand breaks (SSB) and double-strand breaks (DSB) were quantified by performing the alkaline comet assay, and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. DSBs were quantified by phospho-histone 2AX-p53-binding protein 1 co-localization DSB focus assay. The data were compared with cell death quantification by the caspase-3/7 apoptosis assay. Results: Treatment with NIPP led to exceedingly rapid damage to genomic DNA and the appearance of DNA SSBs and DSBs in the initial 4 h. However, damage decreased again within the first 4-8 h, then the late phase began, characterized by DNA DSB and increasing caspase-3/7 activation. Conclusion: Although NIPP treatment leads to extremely rapid damage to genomic DNA, this damage is reversed very quickly by efficient DNA-repair processes. As a consequence, only those cells whose genome damage can be repaired actually survive and proliferate. Persistent genotoxic effects were not observed in the cell system used.

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