P-056 Oxidative stress and human semen quality: new results for diagnostic process of male infertility

Abstract Study question Is oxidative stress (OS) evaluated in human spermatozoa a predictive marker of better semen quality? Summary answer CellROX®Orange is new fluorescent probe able to detect Reactive Oxygen Species (ROS) identifying viable oxidized sperm fraction related performance. What known already OS, defined as an unbalance between ROS production and antioxidant defences, considered one the causes male infertility. OS evaluation represents important goal research this field because parameter could be helpful predict fertilization ability improve Assisted Reproductive Technology outcomes. By using different probes methods for or semen, several studies were performed demonstrating negative role on functions. Such not conclusive small number included subjects, high variability cohorts lack validation. design, size, duration An observational study was conducted 121 samples from patients undergoing routine analysis couple infertility Andrology Laboratory Careggi University Hospital Florence September 2021 March 2022. Washed Swim-up selected incubated with two probes, CellROX® Orange Dihydroethidium (DHE), revealed by flow cytometry. Participants/materials, setting, After analysis, percentage DHE, then correlated standard parameters DNA fragmentation (sDF, TUNEL/PI). Sperm kinematic hyperactivated motility also assessed C.A.S.A. system. Furthermore, positivity phosphatidylserine membrane exposure (determined Annexin V staining) Caspase 3 7 activities (measured FLICA™) concomitantly evaluated. Main results chance We demonstrated that hydrogen peroxide only positive positively associated quality negatively sDF. To confirm these results, we same experiments another which reveals superoxide anion both unviable cells. Similarly Orange, correlation sDF found spermatozoa. Conversely, spermatozoa, opposite associations observed. These indicate status quality. further investigate possibility, double labelled (a early signs apoptosis) well FLICA™ (that detects caspase activity, late sign apoptosis). mostly identifies without apoptotic features. finding significantly higher levels respect unselected samples, once again Limitations, reasons caution Our within there are cells physiological non-physiological intracellular levels, however, they do explain at what cease functional become deleterious Wider implications findings commercially available allows identification Therefore, may useful determine such fraction, likely improving diagnostic process Trial registration applicable