Background and Aims: Infertility is a prevalent health concern, with male factors solely responsible in approximately 20% of infertile couples and contributory in an additional 30% to 40%. In vitro sperm preparation using motility-enhancing agents prior to assist reproductive techniques (ART) has shown promise in improving semen quality and increasing the likelihood of pregnancy. This study aimed to evaluate a novel combination of motility stimulants for improving sperm function, particularly DNA fragmentation index (DFI), and to compare the effectiveness of swim-up and layering techniques for sperm activation.
Method: Semen samples were collected from 90 asthenozoospermic men. Inclusion criteria included normozoospermic fertile men, idiopathic infertile men, and those with oligozoospermia or teratozoospermia. Exclusion criteria included severe oligozoospermia, thyroid dysfunction, current antibiotic therapy, and azoospermia. Samples were divided into two groups: a control group using Ham’s F-12 medium and a treatment group using medium supplemented with Maca powder (1 mg/mL), pentoxifylline (10 mg/mL), and L-carnitine (0.5 mg/mL). Swim-up and layering techniques were used for sperm activation. Parameters were assessed per WHO guidelines (1999 and 2021). Data were analyzed using SPSS version 23.0 with Student t test, ANOVA, and LSD test.
Results: The treatment group showed significant improvements (P<0.05) in sperm concentration, motility (grades A and B), morphology, and reduced DFI. Both preparation techniques yielded comparable enhancements, with layering showing slightly better results in DNA integrity.
Conclusions: The addition of pentoxifylline, L-carnitine, and Maca improved sperm function. The layering method was more effective in selecting spermatozoa with intact DNA.