Increased platelet storage time is associated with mitochondrial dysfunction and impaired platelet function

Abstract

Background Hemorrhagic shock is a leading cause of death following severe trauma and platelet transfusions are frequently necessary to achieve hemostasis. Platelets, however, require special storage conditions and storage time has been associated with loss of platelet quality. We hypothesized that standard storage conditions have a deleterious effect on platelet mitochondrial function and platelet activation. Materials and methods Platelet donations were collected from healthy donors (n=5), and stored in gas permeable collection bags according to American Association of Blood Bank recommendations. Platelet units were sampled from day of collection (day 0) until day 7. High resolution respirometry was used to assess baseline mitochondrial respiration, maximal oxygen utilization, and individual mitochondrial complex-dependent respiration. FACS was performed to analyze mitochondrial content, mitochondrial reactive oxygen species (ROS), the expression of P-selectin (both before and after challenge with thrombin receptor activating peptide (SFLLRN)), and apoptosis. Data was analyzed using ANOVA and Pearson’s correlation (p<0.05 significant). Results Mitochondrial respiration decreased significantly in platelets stored longer than 2 days (p<0.05). Platelets also demonstrated a persistent decrease in response to stimulation with SFLLRN by the 3rd day of storage (p<0.05) as well as an increase in mitochondrial ROS and apoptosis (p<0.05). Mitochondrial respiration significantly correlated with platelet capacity to activate (r=0.8, p<0.05). Conclusion Platelet mitochondrial respiratory function and activation response decrease significantly in platelets stored for 3 or more days. Because platelet transfusions almost universally occur between the 3rd and 5th day of storage, our findings may have significant clinical importance and warrant further in vivo analysis.

Defining the effects of storage on platelet bioenergetics: The role of increased proton leak

“…It has been reported that ATP, ADP and AMP levels decrease during storage suggesting a deficiency in glycolysis and/or oxidative phosphorylation [11]. A number of studies have reported decrease in glucose levels, and an increase in lactate, in the platelet storage bags, suggesting an active glycolytic pathway and possibily inhibition of mitochondrial function [12–14]. Since this increase in glycolytic rate occurs despite an increase in fatty acids, which we would expect could support mitochondrial function, it also suggests a possible mitochondrial defect.…”

Reactive Oxygen Species Generated by CD45- Cells Distinct from Leukocyte Population in Platelet Concentrates Is Correlated with the Expression and Release of Platelet Activation Markers during Storage

“…The authors were also able to show that platelet aggregation increased in proportion to the free radical generation during storage, which indicated that ROS generation is correlated with platelet aggregation. In another study [38] performed on PRP obtained from healthy volunteers, platelet storage time was shown to be associated with elevated generation of mitochondrial-based ROS as well as a progressive increment in apoptosis detected by platelet mitochondrial respiration and functional decline. This study indicated a significant increase in Fig.…”

Biologicals, platelet apoptosis and human diseases: An outlook

“…It is the basis of PRP therapy for tissue repair and it is being used in the treatment of osteoarthritis, bone repair and regeneration, tendinitis, nerve injury, cardiac muscle injury, etc. However, it has been reported that in stored PRP even under standard banking conditions, platelets eventually lose their activity and tend to undergo apoptosis [57]. Thus, it is obvious that there will be an increased concentration of MPs in stored platelet concentrates, which might lead to secondary complications if used for treatment.…”