Platelet Aggregation, Mitochondrial Function and Morphology in Cold Storage: Impact of Resveratrol and Cytochrome c Supplementation

Abstract

Donated platelets are critical components of hemostasis management. Extending platelet storage beyond the recommended guidelines (5 days, 22 °C) is of clinical significance. Platelet coagulation function can be prolonged with resveratrol (Res) or cytochrome c (Cyt c) at 4 °C. We hypothesized that storage under these conditions is associated with maintained aggregation function, decreased reactive oxygen species (ROS) production, increased mitochondrial respiratory function, and preserved morphology. Donated platelets were stored at 22 °C or 4 °C supplemented with 50 μM Res or 100 μM Cyt c and assayed on days 0 (baseline), 5, 7 and 10 for platelet aggregation, morphology, intracellular ROS, and mitochondrial function. Declining platelet function and increased intracellular ROS were maintained by Res and Cyt c. Platelet respiratory control ratio declined during storage using complex I + II (CI + CII) or CIV substrates. No temperature-dependent differences (4 °C versus 22 °C) in respiratory function were observed. Altered platelet morphology was observed after 7 days at 22 °C, effects that were blunted at 4 °C independent of exposure to Res or Cyt c. Storage of platelets at 4 °C with Res and Cyt c modulates ROS generation and platelet structural integrity.

Platelet mitochondria, a potent immune mediator in neurological diseases

“…However, high-dose clopidogrel inhibits mitochondrial respiration by reducing mitochondrial oxidative phosphorylation (Zahno et al, 2013). During platelet storage, supplementation with resveratrol and cytochrome c maintained platelet aggregation, morphology, intracellular ROS, and mitochondrial function (Ekaney et al, 2022). Trehalose is an ideal agent for platelet storage because it improves the apoptosis, viability, and survival rate of platelets at cold temperatures (Baghdadi et al, 2022).…”

Increased platelet mitochondrial function correlates with clot strength in a rodent fracture model

“…Indeed, mitochondrial dysfunction has been implicated in multiple pathologic systemic processes, such as acute respiratory distress disorder and sepsis, 13,14 and has also been used to specifically evaluate the functional properties of donated platelets under various storage conditions. 15,16 Analysis of mitochondrial bioenergetics with high-resolution respirometry (HRR) allows for direct assessment of aerobic respiration, with specific substrate and inhibitor protocols allowing for sensitive analysis of each electron transport chain subunit. A thorough understanding of platelet mitochondrial bioenergetics after injury may provide important clinical prognostic information, more sensitively identify effects of antiplatelet agents, and suggest new targets for platelet-focused antithrombotic therapies.…”

“…Measurement of mitochondrial respiration in circulating platelets may be a useful assay focusing on agonist-independent platelet activation potential. Indeed, mitochondrial dysfunction has been implicated in multiple pathologic systemic processes, such as acute respiratory distress disorder and sepsis, 13,14 and has also been used to specifically evaluate the functional properties of donated platelets under various storage conditions 15,16 . Analysis of mitochondrial bioenergetics with high-resolution respirometry (HRR) allows for direct assessment of aerobic respiration, with specific substrate and inhibitor protocols allowing for sensitive analysis of each electron transport chain subunit.…”