Optimal Concentration of Thrombin to Activate Platelet for Wound Healing

Eum, Soo Jin; Han, Seung Kyu; Chun, Kyung Wook; Kim, Woo Kyung

Abstract

PURPOSE
Platelet transplantation is a novel therapeutic strategy for acceleration of wound healing. When applying platelets, efficacy of adding thrombin to stimulate growth factor release from platelets has already been proven. However, no quantitative data of the thrombin treatment has been reported. The purpose of this study is to determine the optimal thrombin concentration to maximize growth factor release of platelets. In particular, this study was designed to quantify levels of platelet derived growth factor(PDGF)-BB, which is a major growth factor containing in the platelets, in vitro.
METHODS
Fresh platelets were obtained from a blood bank. They were suspended in DMEM/F-12 and incubated with thrombin of various concentrations. The concentrations of thrombin tested were 0, 6.25, 12.5, 25, 50, 100, 200, and 400IU/mL. After 30 minutes, 1, 3, 5, and 7 days, the levels of PDGF-BB were measured using enzyme linked immunosorbent assay. Platelets from four donors were included in this study. Each sample was tested in triplicate and the mean value was used as a data for each sample.
RESULTS
The addition of thrombin increased the level of PDGF-BB. Increases in storage time of platelets resulted in decreased levels of PDGF-BB. Higher levels of PDGF were detected in consort with increased thrombin concentrations. However, there was no significant difference between samples of 200 and 400IU/mL concentrations.
CONCLUSION
The results indicate that adding thrombin accelerates the release of growth factors from platelets and the optimal thrombin concentration to maximize this function is 200IU/mL.