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

## TL;DR

Adding a low dose of platelet-rich plasma before freezing improves goat sperm quality after thawing.

## Contribution

Optimized PRP pre-incubation at 10 million platelets/mL enhances post-thaw sperm quality in goats.

## Key findings

- Pre-incubation with 10 million platelets/mL PRP improved sperm motility and reduced DNA damage.
- Higher PRP doses (20 and 40 million/mL) did not provide additional benefits.
- PRP supplementation in the extender improved some parameters but not oxidative stress or DNA fragmentation.

## Abstract

Freezing goat semen is widely used in animal breeding, but the freezing and thawing process can seriously damage sperm cells, reducing their movement and overall quality. Platelet-rich plasma (PRP) is a natural biological product rich in growth factors and antioxidants, and it has recently attracted attention for its potential protective effects on sperm cells. In this study, we investigated whether adding PRP to goat semen before freezing could protect sperm quality after thawing. Semen samples from Boer goats were treated with different PRP doses (10, 20, and 40 million platelets per milliliter) using two different methods: direct pre-incubation of sperm with PRP and addition of PRP to the freezing solution. Sperm movement, cell membrane health, energy production, oxidative stress, and DNA damage were evaluated after thawing. The results showed that pre-incubating sperm with a low dose of PRP (10 million platelets/mL) before freezing provided the best protection. This treatment improved sperm movement, including progressive motility, preserved cell membranes and mitochondrial activity, and reduced oxidative stress and DNA damage. Higher PRP doses were not more effective. These findings suggest that using PRP in an optimized and carefully applied manner can help improve the success of goat breeding programs.

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 × 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 × 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 × 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 × 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 × 106 platelets/mL as the most effective approach for enhancing post-thaw sperm quality in goats.