Optimization of pure platelet-rich plasma preparation: A comparative study of pure platelet-rich plasma obtained using different centrifugal conditions in a single-donor model

Abstract

While it has been proved that centrifugal conditions for pure platelet-rich plasma (P-PRP) preparation influence the cellular composition of P-PRP obtained, the optimal centrifugal conditions to prepare P-PRP have not yet been identified. In the present study, platelet-containing plasma (PCP) was prepared with the first-spin of different double-spin methods and P-PRP was prepared with different double-spin methods. Whole-blood analysis was performed to evaluate the cellular composition of PCP and P-PRP. The basal and ADP-induced CD62P expression rates of platelets were assessed by flow cytometry to evaluate the function of platelets in PCP and P-PRP. Enzyme-linked immune sorbent assay was performed to quantify interleukin-1β, tumor necrosis factor-α, platelet-derived growth factor AB and transforming growth factor β1 concentrations of PCP and P-PRP. Correlations between the cellular characteristics and cytokine concentrations of P-PRP were analyzed by Pearson correlation analysis. Effects of P-PRP on the proliferation, survival and migration of human bone marrow-derived mesenchymal stem cells and human articular chondrocytes were evaluated by a Cell Counting Kit-8 assay, live/dead staining and Transwell assay, respectively. The results showed that centrifugation at 160 × g for 10 min and 250 × g for 15 min successively captured and concentrated platelets and growth factors significantly more efficiently with preservation of platelet function compared with other conditions (P<0.05). The correlation analysis showed that the similar leukocyte concentrations and leukocyte-reducing efficiencies resulted in similar pro-inflammatory cytokine concentrations in P-PRP (P>0.05) and the maximization of platelet concentration, platelet enrichment factor, platelet capture efficiency and platelet function resulted in the maximization of growth factor concentrations in P-PRP obtained using the optimal conditions (P<0.05). Compared with P-PRP obtained under other conditions, P-PRP obtained under the optimal conditions significantly promoted the proliferation and migration of cells (P<0.05) and did not alter cell survival (P>0.05). Therefore, centrifugation at 160 × g for 10 min and 250 × g for 15 min successively with removal of the buffy coat as a crucial step may provide an optimal preparation system of P-PRP for clinical application.

Original Effective, Safe Technique of Obtaining Platelet Rich Plasma by Centrifugation of the Blood Plasma in Modified Syringe Containers

“…In previously published studies, the maximum efficiency of platelet concentration was demonstrated in double centrifugation with a 160g×10min + 250g×15min mode [18]. In choosing the centrifugation protocol, they relied on these indicators, adapting the rate of acceleration to the rotor design of the centrifuge used.…”

The Differential Effects of Leukocyte-Containing and Pure Platelet-Rich Plasma on Nucleus Pulposus-Derived Mesenchymal Stem Cells: Implications for the Clinical Treatment of Intervertebral Disc Degeneration

“…2 ml whole blood was used in quantifying the platelet and leukocyte concentrations in whole blood, and the rest 28 ml blood was left for preparation of PRP (P-PRP, L-PRP). The method of two-step centrifugation process [30] was applied to prepare the P-PRP and L-PRP. Briefly, 14 ml whole blood was centrifuged at 250g for 10 minutes at room temperature to separate the blood into three layers, platelet-containing plasma at the top, buffy coat (rich in leukocytes and platelets) in the middle, and erythrocytes at the bottom.…”

Importance of homogenization in platelet-rich plasma researches

“…We used the ACD-A tubes to make homemade PRP s after evaluating the various protocols published in the literature [1][2][3][4][5][6][7][8][9][10][11][12]. There are many protocols for PRP and different concentrations are reported while using different spin times and different g forces.…”

“…Sampling from the platelets collected on the buffy coat determines the number of platelets in the PRP [1]. However, having similar platelet counting throughout the PRP is important for treatment.…”

“…During PRP preparation, it is generally known that PRP is confined to a small area immediately on the buffy coat [1]. Sampling from the platelets collected on the buffy coat determines the number of platelets in the PRP [1].…”