Manual platelet-rich plasma production in donkeys by double centrifugation results in leukocyte-rich platelet-rich plasma compared to single centrifugation

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Objective: To compare asinine platelet-rich plasma (PRP) manually produced by single- and double-centrifugation methods. Methods: This was a single-center study conducted from June 19 through August 14, 2022, using 6 healthy donkeys. Whole blood (WB) was collected into sodium citrate vacutainer tubes for single-centrifugation processing and an acid-citrate-phosphate-dextrose-adenine blood bag for double-centrifugation processing to produce, respectively, PRP1 and PRP2. Platelet, WBC, and RBC concentrations and PDGF-BB and TGF-β1 activities were assessed in WB, PRP1, and PRP2. Results: Both protocols concentrated platelets (PLTs) 1.8- to 5.2-fold, reduced WBCs 1.1- to 50.4-fold, and decreased RBCs at least 829-fold compared to WB. Platelet-rich plasma-2 yielded a higher PLT concentration and PLT enrichment factor than PRP1 but required resuspension of the pellet post second spin to maximize PLT concentration. Platelet-rich plasma-1 possessed a lower WBC concentration and greater WBC reduction factor than PRP2. Platelet-derived growth factor-BB and TGF-β1 activities were highest in PRP2 and not significantly different between PRP1 and WB. There was a weak and moderate correlation of baseline PLT concentration to that of PRP2 (r = 0.4) and PRP1 (r = 0.62), respectively; neither was statistically significant. Platelet-rich plasma-2 yielded higher PLT enrichment and growth factor activities despite greater WBC and RBC contamination than PRP1. Conclusions: In donkeys, double centrifugation results in leukocyte-rich PRP with a higher PLT concentration compared to leukocyte-poor PRP with a lower PLT concentration yielded from single centrifugation. Clinical Relevance: Asinine PRP can be manually prepared. While this improves cost-efficient field management of donkeys often treated under resource limitations, the optimal cellular composition, in vivo efficacy, and safety of asinine PRP warrant further investigation.

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