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espanolObjetivo. El plasma rico en plaquetas (PRP) es utilizado de forma cada vez mas frecuente en tecnicas quirurgicas de regeneracion tisular. No obstante, el procesamiento de la sangre hasta obtener PRP puede desen cadenar la activacion prematura de las plaquetas y la perdida de los factores bioactivos. En este trabajo estudiamos la calidad de los concentrados de plaquetas obtenidos siguiendo la tecnica de doble centrifugacion en tubo. Metodo. Se someten 50 ml de sangre a una primera centrifugacion a 200g 10 minutos, se recoge el sobrenadante y se centrifuga a 700g 15 minutos. Posteriormente, tras eliminar las 2/3 partes del plasma, se resuspenden las plaquetas y se analiza el grado de enriquecimiento, el estado de activacion y la reserva funcional de las plaquetas. Resultados. El enriquecimiento en plaquetas del PRP fue de 364?1 77% (n=45) respecto de los niveles presentes en sangre total. Mediante el estudio de la expresion de CD62 por citometria de flujo se determino el porcentaje de plaquetas activadas en las muestras de 8 donantes. Mientras que en la sangre no procesada se detecto un 2, 7% de plaquetas activadas, tras la preparacion del PRP este era solo de 3, 6%, aumentando hasta el 16% en el concentrado almacenado toda la noche a 22° C. Tras la estimulacion con trombina el porcentaje de plaquetas activadas fue de 96, 2%. Conclusion. Este protocolo de preparacion de PRP no produce una activacion significativa de las plaquetas. La respuesta a la estimulacion con trombina de los con centrados indica un buen estado de reserva plaquetaria. EnglishObjetive. Platelet Rich Plasma (PRP) is an autologous preparation currently used in oral maxillofacial recosntructive surgery. Blood collection and preparation of platelet concentrates may lead to platelet activation and the premature loss of their granular load. In this study, we have analyzed the quality of the PRP obtained from a small volume of whole blood through a double centrifugation technique, so called "tube method". Design. We obtained 50 ml of whole blood from 45 patients and centrifuged at 200 g for 10 minutes. The plasma and buffy.coat were collected and we then centrifuged at 700 gr for 15 minutes. The pellet wasresuspended after discarding 2/3 of the plasma. The platelet concentration, platelet activation and the functional response to thrombin were analyzed in these samples. Results. By using this method of PRP preparation, were obtained a 364 +- 177% increase in platelet concentration in comparision with whole blood levels. Platelet activation, measured by flow cytrometry analysis of CD62 expression, was of 2.7% in unprocessed blood and 3.6% in fresh PRP. This figure increased to 16% in PRP samples after overnight storage at room temperature. A percentage of 96% of platelets showed activation in PRP samples after thrombin stimulation. Conclusion. Our results show that platelets contained in PRP concentrates obtained by this method are not significantly activated. A good functional platelet reserve is preserved through the procedure, since platelets maintained a satisfactory response to thrombin after PRP preparation.

Calidad del plasma rico en plaquetas: estudio de la activación plaquetaria