Effect of Plasma-rich in Growth Factors Supplementation Timing on Cryopreservation Outcomes in Normozoospermic Samples and Its Protective Role in Asthenoteratozoospermia

Background: Sperm cryopreservation may have detrimental effects on sperm parameters freezing and thawing. Aim: In this study, the effects of plasma-rich in growth factors (PRGFs) at different stages of freezing-thawing on sperm parameters in normozoospermia and asthenoteratozoospermia (AT) were evaluated. Settings and Design: Aprospective experimental study conducted in two phases to evaluate the timing of PRGF addition during sperm freezing and to assess the cryoprotective effects of PRGF on AT samples. Materials and Methods: In the first phase, 20 normal semen samples were included in the study. After preparation, the spermatozoa were divided into the following groups: A control group that was frozen and thawed without PRGF, and three groups that received 1% PRGF before freezing, during equilibration and after thawing. The method of cryopreservation was rapid freezing. Sperm motility, viability, normal morphology, deoxyribonucleic acid (DNA) fragmentation, reactive oxygen species (ROS) levels and mitochondrial membrane potential were evaluated in different groups before freezing and after thawing. In the second phase, 1% PRGF was added to 10 AT samples, and the parameters were evaluated as in the first stage and compared with the control group. Statistical Analysis Used: test were applied as appropriate. Results: In the first phase, progressive motility in all groups was significantly reduced after thawing compared to fresh spermatozoa. Furthermore, in the group that received PRGF during equilibration, the sperm total motility and viability increased significantly compared to the control group. The rate of DNA fragmentation and ROS also increased significantly in all groups, except in the during equilibration group. In the second phase, the group treated with 1% PRGF after thawing showed no significant differences in total motility, viability, ROS or DNA integrity compared to the fresh spermatozoa. Conclusion: Adding 1% PRGF to the freezing medium during the equilibration process yields effective results. PRGF effectively preserves sperm motility, viability and DNA integrity in AT samples.