Oxidative alterations during human platelet storage

Abstract

SUMMARY: During storage of platelet obtained by apheresis several changes occur. The aim of this study was to investigate the effect of storage on activation, apoptosis, protein pattern, lipid peroxidation, and the levels of nitric oxide (NO) and glutathione (GSH) of platelets. In this study, platelets obtained from healty donors (n=7) by apheresis were kept in an agitator for nine days at 20-24°C. The samples were taken on the 1st, 3 rd, 5 th and 9 th days and platelets were precipitated. Platelet activation with PAC-1 and CD62-P antibodies and platelet apoptosis were measured with Annexin-V using flow cytometer. Platelets were frozen and thawed four times and then NO, GSH and malondialdehyte (MDA) levels were assayed by spectrophotometry. Platelets protein pattern was investigated via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) procedure. When compared to the 1st day, platelet CD62-P, PAC-1 expressions and Annexin-V levels significantly increased on the 3rd, 5th and 9th days, while platelet NO and GSH levels significantly decreased on the 3rd, 5th and 9th days. Furthermore MDA levels significantly increased on only the 5th and 9th days. Mild changes occurred in the density of platelet protein bands. In conclusion, our results show that alterations of platelet activity during storage period may enhance platelet procoagulant activity which increases trombogenic risk. Therefore, for transfusion using fresh platelets or adjusting platelet preservation are strongly important for platelet behavior in vivo conditions.

The effect of gamma irradiation on platelet redox state during storage

“…We also demonstrated significantly lower levels of GSH in gamma‐irradiated PLTs on Days 0, 1, and 3 of storage. Our findings about storage‐dependent decreases in intracellular GSH were in accordance with previous studies 58,62,65 . However, to the best of our knowledge this is the first report that indicated that gamma irradiation decreases intra‐PLT GSH in stored PCs.…”

“…However, they also indicated that, with the significant induction of oxidative state during storage the levels of both GSH and GSSG in 5‐day‐stored PCs reach the comparable amount for both gamma irradiated and untreated controls 29 . Regardless of irradiation effects, several other studies also used GSH levels to monitor PLTs redox state during storage; nonetheless, they evaluated intra‐PLTs GSH rather than the released one, 58,62–65 which may not be an appropriate method to evaluate intracellular redox state. Similar to others here, we examined the intracellular GSH while in both irradiated and nonirradiated PCs gradual decreases of GSH were indicated during storage.…”

Evaluation of store lesion in platelet obtained by apheresis compared to platelet derived from whole blood and its impact on the in vitro functionality

“…1). This result is consistent with the a study of Goker et al in 2011 where seven platelet concentrates obtained by apheresis showed a significant difference in the concentration of malondialdehyde (MDA) between days 5 and 7 of storage, attributing it mainly to damage caused by ROS to polyunsaturated fatty acids of the membrane [27].…”

Influence of Oxidative Stress on Stored Platelets

“…OS leads to decrease in platelet nitric oxide which increases platelet activation and cellular production of reactive oxygen species (ROS) [ 4 ]. But whether resulting oxidative changes trigger activation and apoptosis or activation and apoptosis trigger oxidative changes during storage is still unclear [ 5 ].…”