Abstract
Background: Platelet rich plasma (PRP) is used extensively in equine regenerative medicine. Differences in preparation protocols give rise to significant variability in the cellular composition of PRP making it very difficult to establish a standard of care in the field. This study aimed to optimize the preparation protocol for leukocyte-reduced PRP (P-PRP).Methods: Blood (100 mL) was collected from horses (n = 5) and divided into 2 purple top EDTA tubes and 6 (15 mL) double syringesa with a final concentration of 10% acid citrate dextrose anticoagulant. Six double syringesa were collected from each horse; PRP samples were prepared in duplicate and centrifuged at 1,100 rpm (188 × g), 1,300 rpm (263 × g), or 1,500 rpm (350 × g). Duplicates were subjected to +/– braking at the end of centrifugation. The total volume of PRP generated was measured and divided into thirds. Each third (top, middle, and bottom) were drawn off separately using the inner (6 mL syringe) and placed in purple top EDTA tubes. Automated complete blood counts were performed on all peripheral whole blood and PRP samples.Results: The concentration of leukocytes was higher in the bottom layer of PRP compared to the top and middle layers (p < 0.0001). The concentration of platelets was slightly lower in the bottom layer of PRP than the middle layer (p = 0.02). Centrifugation braking increased the leukocyte concentration in the top (p = 0.03) and middle layers of PRP (p = 0.001). Centrifugation rate had no effect on the cellular composition of PRP (p = 0.1–0.6).Conclusions: Because layer of plasma affected both platelet and leukocyte concentrations in PRP, the most important modification for the current single spin, double syringe, plasma based PRP preparation protocols is to exclude the bottom 1/3 layer of PRP.
Highlights
Musculoskeletal injuries are common in equine athletes and, due to the poor intrinsic healing capabilities of cartilage and tendons, present a significant clinical challenge [1, 2]
A total of 95 Complete blood counts (CBCs) were performed on 30 Platelet rich plasma (PRP) and 5 peripheral whole blood samples
Platelet concentration was affected by the layer of PRP, but not by centrifugation rate or braking
Summary
Musculoskeletal injuries are common in equine athletes and, due to the poor intrinsic healing capabilities of cartilage and tendons, present a significant clinical challenge [1, 2]. There is considerably more robust evidence for the application of PRP in musculoskeletal injury with several level one studies supporting the use of PRP for tendinopathies and osteoarthritis [7,8,9,10,11,12,13,14,15,16]. Some studies suggest a beneficial effect including increased growth factor and cytokine release as well as increased antibacterial and immunologic resistance [17]. Other studies suggest a negative effect including increased inflammatory cytokines and catabolic mediators release leading to the degradation of tendon, ligament, muscle, and cartilage [18, 19]. This study aimed to optimize the preparation protocol for leukocyte-reduced PRP (P-PRP)
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