Comparison of reproductive outcomes in ICSI cycles after sperm selection techniques in advanced paternal age

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Background: Advanced paternal age is associated with declining semen parameters, increased sperm DNA fragmentation (SDF), and oxidative stress, which may negatively affect ART outcomes. Microfluidic sperm sorting has been proposed as a method to reduce SDF and improve sperm quality compared to conventional density gradient centrifugation. Methods: Single-center retrospective cohort study including 193 ICSI cycles performed between January 2023 and December 2024. Couples were stratified by male age (<40 years vs. ≥40 years) and further subdivided by sperm preparation method (density gradient vs. microfluidics), forming four subgroups (A1: <40/DG, n=56; A2: <40/MF, n=59; B1: ≥40/DG, n=33; B2: ≥40/MF, n=45). Inclusion criteria: ≥10 mature oocytes, ICSI, frozen double blastocyst transfer, and sperm concentration ≥5×106/ml. Exclusion criteria: severe male factor infertility or donor gametes. Primary outcome: live birth rate. Secondary outcomes: clinical pregnancy, miscarriage, fertilization, and blastulation rates. Results: No significant differences were observed in clinical pregnancy, miscarriage, or live birth rates between preparation methods within either age group (all p>0.05). Fertilization and blastulation rates were comparable across subgroups. A modest reduction in oocyte yield was noted in older males undergoing microfluidic preparation (A2 vs. B2; p=0.034), likely reflecting female- or cycle-specific factors. Demographic/clinical data are detailed in tables in the results section. Multivariate logistic regression showed that female age (OR 1.02, 95% CI 0.96–1.09, p=0.50) and subgroup membership were not significant predictors of clinical pregnancy. Pairwise comparisons with Tukey adjustment confirmed no significant differences between subgroups (e.g., A1 vs. B1: OR 1.91, p=0.515). conclusion: Male age ≥40 years and sperm preparation method (microfluidics vs. density gradient) were not independent determinants of ART success in this cohort. Clinical decision-making regarding sperm selection should not be guided solely by paternal age or subgroup classification.

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