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Assessment of Spermatic Chromatin Decondensation by the Toluidine Blue Assay in Infertile Patients in Cotonou

YauDe Alfred, Azonbakin Simon, Aweh Bruno, Dangbemey Patrice, Sissoko Sidi Boula, Dossou Jean-Paul, Flore Gangbo, Laleye Anatole

Abstract: Background: A high level of sperm chromatin decondensation results in loss of DNA quality and it is associated with poor embryonic prognosis in Assisted Reproductive Technology (ART). This had lead to introduction of sperm DNA compaction testing in assessment of clinical male infertility. Toluidine blue assay is one of most commonly used tests to measure chromatin decondensation in research studies and also in clinical routine use. So the objective of our study was to assess the level of sperm chromatin decondensation by toluidine blue assay in men dealing with couple infertility. Methods: The sperm samples of 96 patients referred for infertility, over a 4-month period from September 2020 to January 2021, were stained with toluidine blue and observed under an optical microscope. These were evaluated according to the World Health Organization. Results: The 25 - 34 age group was the most represented among infertile men. There was a non-significant correlation between age and the level of spermatic chromatin condensation (r = 0.0348). Spermatic parameters had averages above standards norms, apart from that of low vitality. Spermatic chromatin decondenization affected 96% of infertile patients. Asthenozoospermia was the most common abnormality. Among standard parameters of spermogram, only the concentration of spermatozoa in the semen was weakly correlated (r = 0.18) with sperm chromatin decondensation. Conclusion: The level of sperm chromatin condensation is, on the one hand, a parameter independent of the age of the subject; on the other hand, allows to evaluate male infertility, like the classic parameters of the spermogram and spermocytogram. However, in view of the high probability of changes in the nuclear quality of sperm intrinsic to the analysis process and sperm concentration, an improvement of the toluidine blue test using the swim-up technique and associated with image processing software is possible.
Keywords: Male Infertility / Decondensation / Spermatic Chromatin / Toluidine Blue

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