Introduction. Platelet-Rich Plasma (PRP) is rich in growth factors, playing important role in tissue healing. The wide variation of reported protocols for preparation of PRP leads to variable compositions, which induce different biological responses and prevent results comparison. This study aims to highlight relevant aspects of the centrifugation step to obtain reproducible results and overall quality. Material and Methods. Samples of blood were collected from 20 healthy donors that have signed free informed consent. Two centrifugation steps (spins) were analyzed for the influence of centrifugal acceleration, time, processed volume, and platelet gradient. The Pure Platelet-Rich Plasma (P-PRP) was characterized as platelet concentration, integrity, and viability (sP-selectin measurement). Results. Lower centrifugal accelerations favour platelet separation. The processing of 3.5？mL of blood at 100？×g for 10？min (1st spin), 400？×g for 10？min (2nd spin), withdrawing 2/3 of remnant plasma, promoted high platelet recovery (70–80%) and concentration (5x) maintaining platelet integrity and viability. The recovery of platelets was reduced for a larger WB volume (8.5？mL) processed. Conclusion. Centrifugal acceleration, time, WB processed volume, and minimization of the platelet gradient before sampling are relevant aspects to ensure reproducible compositions within the autologous nature of PRP. 1. Introduction Platelet-Rich Plasma (PRP) is an autologous preparation that concentrates platelets in a small volume of plasma [1]. Platelets are rich in growth factors, which play an important role in tissue healing. Numerous studies have demonstrated the clinical application and notable results of PRP in dentistry [2], oral maxilla facial surgery [3], plastic surgery [4], orthopedics [5], rheumatology [6], and the treatment of different types of injuries that include chronic wounds [7, 8] and muscle injuries [9]. PRP is made for two purposes: one for harvesting platelets for therapeutic purposes and the other for testing for platelet function in PRP using aggregometry. In this work it was studied for therapeutic purposes only. The wide variation in the reported protocols for obtaining PRP may lead to samples with different compositions that may induce different biological responses [1]. Despite these variations, all protocols follow a generic sequence that consists of blood collection, an initial centrifugation to separate red blood cells (RBC), subsequent centrifugations to concentrate platelets, and other components and an activation of the sample by adding a
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