Zeta and hyaluronic acid assessments, novel sperm selection procedures, in animal model for male infertility.

Considering varicocele (VCL)-induced severe, progressive DNA damage, histone-protamine anomalies and low sperm production, in the current study, the experimental VCL was induced and the efficiency of hyaluronic acid (HA)-binding method (HABM) and zeta preparation procedure (ZPP) in selection of appropriate spermatozoa was compared with those spermatozoa from intact animals. Following 2 and 4 months, the histological alterations in testicular tissue, sperm count and viability were assessed to prove the VCL condition. The spermatozoa were undergone simple wash, HABM and ZPP. The chromatin condensation, active caspase-3 expression, DNA fragmentation and apoptosis index were analysed after applying selection techniques and compared with the spermatozoa from intact and VCL-induced animals, which were undergone a simple wash. Observations showed that both HABM and ZPP effectively prepared the spermatozoa with higher chromatin condensation and lower DNA damage. Meanwhile, the ZPP exerted a more preferable effect by preparing the spermatozoa with higher chromatin condensation, and lower caspase-3 expression, and DNA disintegrity versus the HABM, especially after 4 months. In conclusion, ZPP seems to exert much more reliable efficiency in selecting appropriate spermatozoa for ICSI processes, while more studies are needed to find out which one is more useful in the clinical assisted reproductive technique (ART) process.