Amniotic Mesenchymal-Derived Extracellular Vesicles and Their Role in the Prevention of Persistent Post-Breeding Induced Endometritis

Persistent post-breeding induced endometritis (PPBIE) is considered a major cause of
subfertility in mares. It consists of persistent or delayed uterine inflammation in susceptible mares.
There are many options for the treatment of PPBIE, but in this study, a novel approach aimed at
preventing the onset of PPBIE was investigated. Stallion semen was supplemented with extracellular
vesicles derived from amniotic mesenchymal stromal cells (AMSC-EVs) at the time of insemination to
prevent or limit the development of PPBIE. Before use in mares, a dose–response curve was produced
to evaluate the effect of AMSC-EVs on spermatozoa, and an optimal concentration of 400 × 106 EVs
with 10 × 106 spermatozoa/mL was identified. At this concentration, sperm mobility parameters
were not negatively affected. Sixteen susceptible mares were enrolled and inseminated with semen
(n = 8; control group) or with semen supplemented with EVs (n = 8; EV group). The supplementation
of AMSC-EVs to semen resulted in a reduction in polymorphonuclear neutrophil (PMN) infiltration
as well as intrauterine fluid accumulation (IUF; p < 0.05). There was a significant reduction in
intrauterine cytokine levels (p < 0.05) for TNF-α and IL-6 and an increase in anti-inflammatory IL-10
in mares in the EV group, suggesting successful modulation of the post-insemination inflammatory
response. This procedure may be useful for mares susceptible to PPBIE.