Platelet activation and IL-8 production in platelet concentrates prepared by buffy coat and PRP method

ABSTRACT

The process of platelet concentrate production by plasma rich [PRP] method could activate the platelet and granules secretion of beta thromboglobulin, LDH and CD62P. Platelets activated during the preparation process do not have sufficient efficiency for hemostasis in vivo. It seems that platelet preparation by buffy coat method has an ability less than PRP to activate the platelet. Measuring platelet activation indices, such as CD62P expression and beta thromboglobulin, is a useful means to evaluate the percentage of activated platelet concentrates and compare the two methods of buffy coat and PRP. In this experimental study, 15 concentrates were prepared via PRP method and 15 via BC method; 15 intact blood units were also considered as control group. The percentages of CD62P expression, soluble CD62P concentrates, IL-8 level, and CD14 positive cells were evaluated. Special monocolonal antibodies that conjugated with flourecence dye in flocytometric method were used for CD62P and CD14. ELISA method was used for evaluation of soluble CD62P and IL-8. The average platelet count in both methods showed no significant difference, but WBC contamination rate in PRP-PCs was more than BC-PCs. In PRP-PCs, we found a little decrease in CD62P expression and increase in soluble form and IL-8 level during reservation time. The level of CD14 showed no significant difference in these components. In BC method during the three day reservation, expression of CD62P, its soluble form, and IL-8 concentrates increased and the level of monocyte surface CD14 showed slight decrease ranging from 0.4 to 0.1. It is concluded that there is a close relationship between IL-8 and WBC count in platelet concentrates. In PRP method in contrary to BC method, high speed centrifuge causes adhesion, aggregation and platelet activation