Effects of Platelet-Rich Plasma Dose and Application Strategy on Post-Thaw Spermatological Parameters in Goat Semen

Abstract

This study investigated the effects of different PRP doses and application strategies on sperm quality in goats during cryopreservation. Semen samples were collected weekly from six Boer goats and treated with PRP either by means of pre-incubation or supplementation of the semen extender at concentrations of 10, 20, and 40 & times; 106 platelets/mL. Computer-assisted sperm analysis (CASA) was performed after equilibration and again after thawing to assess sperm motility parameters, including progressive motility. Post-thaw sperm functional integrity was further evaluated using flow cytometry, including assessments of plasma membrane integrity, acrosome integrity, high mitochondrial membrane potential (hMMP), sperm viability, oxidative stress, and DNA integrity. The results demonstrated that pre-incubation with PRP at 10 & times; 106 platelets/mL (pre-PRP10) yielded the most pronounced improvements in total and progressive motility after thawing, as well as in plasma membrane integrity, viability, and mitochondrial activity, while significantly reducing oxidative stress and DNA fragmentation compared with other PRP doses and application strategies. Supplementation of the extender with PRP at 20 & times; 106 platelets/mL also showed favorable effects on functional sperm parameters; however, it did not result in significant improvements in oxidative stress or DNA fragmentation. In contrast, the highest PRP dose (40 & times; 106 platelets/mL) did not confer additional benefits. None of the PRP treatments improved post-thaw acrosome integrity. In conclusion, these findings highlight the importance of PRP dose optimization and direct sperm-PRP interaction, identifying pre-incubation with PRP at 10 & times; 106 platelets/mL as the most effective approach for enhancing post-thaw sperm quality in goats.