Abstract
Simple SummaryPlatelet-rich plasma (PRP) is used to improve the regenerative capacity of damaged tissues in different species. In equine medicine, PRP is commonly used to treat joint diseases, tendinitis, ligamentous lesions, and persistent endometritis. Jenny endometrium shows a high sensitivity to endometritis. There are important differences between donkey and horse blood characteristics. Several protocols to obtain horse PRP have been reported, but no protocols have yet been reported for obtaining donkey PRP. Our study shows that single-spin tube centrifugation at 133× g for 10 min is appropriate to obtain donkey PRP with therapeutic potential in jenny endometritis-endometriosis.The aim of this study was to standardize a simple, manual platelet-rich plasma (PRP) protocol in Catalonian donkeys using single-spin tube centrifugation as a treatment for jenny endometritis. The objective was to obtain a blood product with a moderate concentration of platelets (2 or 3 times baseline physiologic values) and a low WBC (White Blood Cells) concentration. Blood was drawn from six Catalonian donkeys using acid citrate dextrose (ACD) as an anticoagulant, and then processed by single centrifugation at 133× g for two different centrifugation times (10 and 15 min). The PRP samples were evaluated by flow cytometry, and TGF-β1 (Transforming Growth Factor-Beta1) concentrations were determined by enzyme-linked immunosorbent assay (ELISA). The 10 min centrifugation protocol resulted in a slightly greater release of TGF-β1 (6044.79 ng/mL), a 2.06-fold increase in platelet concentration, and a 15-fold reduction in leukocyte concentration when compared to the initial values. The 15 min centrifugation time resulted in a 2.44-fold increase in baseline platelet concentration, a reduction in WBC count by a factor of 20, and slightly lower TGF levels (5206 ng/mL). We conclude that both protocols are adequate for the obtention of PRP, and both may have an acceptable therapeutic potential for use in this species, although this needs to be further validated.
Highlights
Jenny endometrium shows high susceptibility to endometritis
Jenny endometritis is characterized by the presence of large amounts of polymorphonuclear neutrophils (PMNs) and eosinophils migrating to the uterine lumen, as well as increased cytokine expression [2,3]
There were no differences in these values (p > 0.05) between the platelet-rich plasma (PRP) prepared with different protocols
Summary
Jenny endometrium shows high susceptibility to endometritis. Semen deposition induces an exacerbated inflammatory response, which is more pronounced than in mares [1,2].Jenny endometritis is characterized by the presence of large amounts of polymorphonuclear neutrophils (PMNs) and eosinophils migrating to the uterine lumen, as well as increased cytokine expression [2,3].After infection, neutrophils destroy pathogens by phagocytosis and by degranulation, producing DNA complexes that combine with nuclear and cytoplasmic proteins to form web-like extracellular structures called neutrophil extracellular traps (NETs) [4]. Jenny endometrium shows high susceptibility to endometritis. Semen deposition induces an exacerbated inflammatory response, which is more pronounced than in mares [1,2]. Jenny endometritis is characterized by the presence of large amounts of polymorphonuclear neutrophils (PMNs) and eosinophils migrating to the uterine lumen, as well as increased cytokine expression [2,3]. After intrauterine semen deposition, the PMN endometrial infiltration is induced by spermatozoa rather than bacteria [5]. A reduced number of spermatozoa are phagocytosed by PMNs, and most remain attached to NETs in a surrounding PMN halo [6]. NETs production is controlled by seminal plasma and represents the main sperm filter [7]
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