Enhanced Platelet Function in Cold Stored Whole Blood Supplemented with Resveratrol or Cytochrome C

ABSTRACTBACKGROUNDLimited availability and use of whole blood (WB) following trauma is driven by perceptions that hemostatic function is limited by platelet dysfunction within 5 days storage. We sought to define the hemostatic function of WB stored at 4°C for up to 25 days, elucidate changes in metabolic parameters and mitochondrial dysfunction in platelets in WB, and the effect of supplementation using resveratrol (Res) or cytochrome c (Cyt c).METHODSWB was collected, aliquoted, and stored at 4°C without agitation. Res or Cyt c was supplemented prior to storage, or 10d post-storage. Serial samples were collected and analyzed for hemostatic function by platelet mapping thromboelastography (PM-TEG). Platelets isolated from WB were counted and mitochondrial function assessed by oxygen consumption, mitochondrial membrane potential, and biochemical parameters.RESULTSPlatelet function of WB was maintained up to 15 days at 4°C before a significant decrease was observed at 25d. Res or Cyt c improved WB aggregation potential when supplemented 10d post-storage. Platelet oxygen consumption was maintained until 10d storage but significantly decreased thereafter in the absence of change in platelet count. Cyt c increased oxygen consumption on day 15 and platelet mitochondrial membrane potential steadily decreased over time, an effect attenuated by Res or Cyt c supplementation 10d post-storage. Potassium and lactate levels increased during storage, while pH levels decreased, with no observed effect following Res or Cyt c supplementation.CONCLUSIONSStoring cold WB with Res or Cyt c supplementation enhances ex vivo aggregation by improving platelet function, thereby extending overall storage life. These findings have potential significance for improving WB availability in immediate trauma situations, including treatment in a battlefield trauma setting.Study typeTranslational studyLevel of evidenceDiagnostic Test or Criteria Level II BACKGROUND Limited availability and use of whole blood (WB) following trauma is driven by perceptions that hemostatic function is limited by platelet dysfunction within 5 days storage. We sought to define the hemostatic function of WB stored at 4°C for up to 25 days, elucidate changes in metabolic parameters and mitochondrial dysfunction in platelets in WB, and the effect of supplementation using resveratrol (Res) or cytochrome c (Cyt c). METHODS WB was collected, aliquoted, and stored at 4°C without agitation. Res or Cyt c was supplemented prior to storage, or 10d post-storage. Serial samples were collected and analyzed for hemostatic function by platelet mapping thromboelastography (PM-TEG). Platelets isolated from WB were counted and mitochondrial function assessed by oxygen consumption, mitochondrial membrane potential, and biochemical parameters. RESULTS Platelet function of WB was maintained up to 15 days at 4°C before a significant decrease was observed at 25d. Res or Cyt c improved WB aggregation potential when supplemented 10d post-storage. Platelet oxygen consumption was maintained until 10d storage but significantly decreased thereafter in the absence of change in platelet count. Cyt c increased oxygen consumption on day 15 and platelet mitochondrial membrane potential steadily decreased over time, an effect attenuated by Res or Cyt c supplementation 10d post-storage. Potassium and lactate levels increased during storage, while pH levels decreased, with no observed effect following Res or Cyt c supplementation. CONCLUSIONS Storing cold WB with Res or Cyt c supplementation enhances ex vivo aggregation by improving platelet function, thereby extending overall storage life. These findings have potential significance for improving WB availability in immediate trauma situations, including treatment in a battlefield trauma setting. Study type Translational study Level of evidence Diagnostic Test or Criteria Level II Address for correspondence Susan L. Evans, MD, FACS, Department of Surgery, Carolinas Medical Center, 1000 Blythe Boulevard, Charlotte, NC, 28203, USA Conflicts of interest statement None of the authors have any conflicts of interest to declare. Previous communication Presented, in part, as a plenary paper at the 9th Annual Military Health System Research Symposium on 08-28, 2017 in Kissimmee, FL. Acknowledgements This study was supported by institutional funds from Carolinas Medical Center. Study type: Translational study Level of evidence: Diagnostic Test or Criteria Level II © 2018 Lippincott Williams & Wilkins, Inc.

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