OBJECTIVE: The aim of this article is to discuss a protocol for obtaining platelet-rich plasma (PRP) and evaluate which factors, derived from its preparation method and from whole blood, modify PRP cytometry and coagulation time. STUDY DESIGN: Whole blood, harvested from 50 rabbits, was centrifuged at 300g for 10 minutes. Supernatant was recentrifuged at 5000g for 5 minutes. PRP was clotted with calcium chloride. Whole blood and PRP cytometry were obtained through automatic measurement. The amount of erythrocyte- and platelet-poor plasma drawn from whole blood was measured. Hematocrit, platelet and leukocyte count, mean corpuscular volume (MCV) and mean platelet volume (MPV), mean, standard deviation, and median were also calculated at whole blood and PRP. PRP coagulation time was also analyzed. Mean values between groups were analyzed using Student t test. Correlations were evaluated using Pearson's correlation coefficient. The significance level was set at P < .05. A linear regression was performed to investigate the relationship among the correlated variables. RESULTS: From whole blood, 2.68 mL of erythrocytes and 5.72 mL of platelet-poor plasma (PPP) were removed. PRP platelet count was 2,324,080 cells/microL. Whole blood hematocrit influenced the amount of cells and PPP removed, as well as PRP platelet count. PRP platelet count was dependent on whole blood hematocrit and platelet count, and does not interfere in PRP coagulation time. A linear interaction was confirmed between the variables that presented significant Pearson correlation. CONCLUSIONS: The protocol evaluated produces a good PRP. Whole-blood parameters can predict PRP features. Whole-blood hematocrit is an important variable for PRP preparation and PRP cytometry characterization. PRP platelet count is dependent upon whole-blood platelet count.
OBJECTIVE: The aim of this article is to discuss a protocol for obtaining platelet-rich plasma (PRP) and evaluate which factors, derived from its preparation method and from whole blood, modify PRP cytometry and coagulation time. STUDY DESIGN: Whole blood, harvested from 50 rabbits, was centrifuged at 300g for 10 minutes. Supernatant was recentrifuged at 5000g for 5 minutes. PRP was clotted with calcium chloride. Whole blood and PRP cytometry were obtained through automatic measurement. The amount of erythrocyte- and platelet-poor plasma drawn from whole blood was measured. Hematocrit, platelet and leukocyte count, mean corpuscular volume (MCV) and mean platelet volume (MPV), mean, standard deviation, and median were also calculated at whole blood and PRP. PRP coagulation time was also analyzed. Mean values between groups were analyzed using Student t test. Correlations were evaluated using Pearson's correlation coefficient. The significance level was set at P < .05. A linear regression was performed to investigate the relationship among the correlated variables. RESULTS: From whole blood, 2.68 mL of erythrocytes and 5.72 mL of platelet-poor plasma (PPP) were removed. PRP platelet count was 2,324,080 cells/microL. Whole blood hematocrit influenced the amount of cells and PPP removed, as well as PRP platelet count. PRP platelet count was dependent on whole blood hematocrit and platelet count, and does not interfere in PRP coagulation time. A linear interaction was confirmed between the variables that presented significant Pearson correlation. CONCLUSIONS: The protocol evaluated produces a good PRP. Whole-blood parameters can predict PRP features. Whole-blood hematocrit is an important variable for PRP preparation and PRP cytometry characterization. PRP platelet count is dependent upon whole-blood platelet count.
Authors: José F S D Lana; Adam Weglein; Steve E Sampson; Eduardo F Vicente; Stephany Cares Huber; Clarissa V Souza; Mary A Ambach; Hunter Vincent; Aline Urban-Paffaro; Carolina M K Onodera; Joyce M Annichino-Bizzacchi; Maria Helena A Santana; William D Belangero Journal: J Stem Cells Regen Med Date: 2016-11-29