Evaluation of effectiveness of different regiments of centrifugation for a double-spin method of pure platelet-rich plasma preparation

Abstract

The purpose - to determine the optimal parameters of double centrifugation in the preparation of pure platelet-rich plasma with a maximum reduction of other blood cellular elements. Materials and methods. The study included 30 apparently healthy individuals aged 18 to 60 years who underwent venous blood sampling. The first centrifugation of all blood samples was performed in the 160 g × 10 min mode. The second centrifugation was performed in 12 different modes: 190 g × 10 min, 190 g × 15 min, 220 g × 10 min, 220 g × 15 min, 250 g × 10 min, 250 g × 15 min, 290 g × 10 min, 290 g × 15 min, 320 g × 10 min, 320 g × 15 min, 360 g × 10 min, 360 g × 15 min. The number of erythrocytes, leukocytes, platelets (in whole blood, after the first and second centrifugation), platelet capture efficiency, platelet enrichment factor, erythrocytes and leukocytes reduction efficiency, concentration of major growth factors in plasma after the second centrifugation were determined. Results. In whole blood, the concentration of erythrocytes was 4.455±0.301 × 1012/l, leukocytes - 6.485±0.501 × 109/l, platelets - 281.5±22.7 × 109/l. After the first centrifugation, the volume of the obtained plasma containing platelets was 4.51±0.25 ml, the number of platelets was 482.1±47.4 × 109/l, with platelet reduction rates of 14.39±1.1%, their preservation - 85.6±1.1% and enrichment factor - 1.71±0.1; the number of erythrocytes was 0.104±0.006 × 1012/l, leukocytes - 0.205±0.025 × 109/l, and their reduction efficiency - 98.83±0.09% and 98.42±0.14%, respectively. After the second centrifugation in different modes, the 250 g × 15 min mode turned out to be the most effective, which made it possible to achieve a platelet enrichment factor of about 4.93 with a concentration of platelets in the final product of 1386.7±116.5 × 109/l, their capture efficiency of 82.1±1.1% and erythrocytes and leukocytes reduction efficiency of 98.86±0.08% and 98.45±0.14%, respectively. Conclusions. In case of double-spin centrifugation, the sequential combination of 160 g × 10 min and 250 g × 15 min regimens is most effective for the preparation of pure platelet-rich plasma. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

The method of correction of hemorrhagic shock action on the pro- and antioxidant system of the periulceral stomach tissues in an experiment

“…In order to simulate hemorrhagic shock, blood was taken from the lateral vein of the tail. The obtained blood was used to prepare platelet-rich plasma (PRP) according to the standard method [14]. The volume of blood was determined individually and was equal to 1.5 % of the animal's weight, which corresponded to a blood loss of 20-25 % of the CBV.…”

“…Platelets are one of the largest sources of growth factors in the body and actively influence tissue healing processes. Thus, platelets contain: platelet-derived epidermal growth factor (PD-EGF), platelet-derived growth factor (PDGF), bone morphogenetic protein (BMP), transforming growth factor (TGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), endothelial cell growth factor (ECGF), basic fibroblast growth factor (bFGF) [5,14,18]. In addition, α-granules of platelets contain adhesive proteins (fibrinogen, fibronectin, vitronectin, thrombospondin-1), coagulation factors (factor V, factor XI, protein S, antithrombin), fibrinolytic factors (plasminogen, urokinase inhibitor, α-2 antiplasmin), proteases and antiproteases (TIMP-4, metalloproteinase-4, α-1 antitrypsin), basic proteins (platelet factor 4, βthromboglobulin, endostatins), membrane glycoproteins (CD40-ligand, P-selectin).…”