Platelet-rich plasma stimulated by pulse electric fields: Platelet activation, procoagulant markers, growth factor release and cell proliferation

Platelets. 2016;27(2):128-35. doi: 10.3109/09537104.2015.1048214. Epub 2015 Jun 1.


Therapeutic use of activated platelet-rich plasma (PRP) has been explored for wound healing, hemostasis and antimicrobial wound applications. Pulse electric field (PEF) stimulation may provide more consistent platelet activation and avoid complications associated with the addition of bovine thrombin, the current state of the art ex vivo activator of therapeutic PRP. The aim of this study was to compare the ability of PEF, bovine thrombin and thrombin receptor activating peptide (TRAP) to activate human PRP, release growth factors and induce cell proliferation in vitro. Human PRP was prepared in the Harvest SmartPreP2 System and treated with vehicle, PEF, bovine thrombin, TRAP or Triton X-100. Platelet activation and procoagulant markers and microparticle generation were measured by flow cytometry. Released growth factors were measured by ELISA. The releasates were tested for their ability to stimulate proliferation of human epithelial cells in culture. PEF produced more platelet-derived microparticles, P-selectin-positive particles and procoagulant annexin V-positive particles than bovine thrombin or TRAP. These differences were associated with higher levels of released epidermal growth factor after PEF than after bovine thrombin or TRAP but similar levels of platelet-derived, vascular-endothelial, and basic fibroblast growth factors, and platelet factor 4. Supernatant from PEF-treated platelets significantly increased cell proliferation compared to plasma. In conclusion, PEF treatment of fresh PRP results in generation of microparticles, exposure of prothrombotic platelet surfaces, differential release of growth factors compared to bovine thrombin and TRAP and significant cell proliferation. These results, together with PEF's inherent advantages, suggest that PEF may be a superior alternative to bovine thrombin activation of PRP for therapeutic applications.

Keywords: Growth factors; platelet-rich plasma; platelets; pulse electric field; wound healing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Annexin A5 / biosynthesis
  • Annexin A5 / genetics
  • Blood Platelets / cytology
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism
  • Cattle
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell-Derived Microparticles / metabolism
  • Electric Stimulation
  • Electricity*
  • Epidermal Growth Factor / biosynthesis
  • Epidermal Growth Factor / genetics
  • Epidermal Growth Factor / metabolism*
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Flow Cytometry
  • Gene Expression
  • Humans
  • Octoxynol / pharmacology
  • P-Selectin / biosynthesis
  • P-Selectin / genetics
  • Platelet Activation / drug effects
  • Platelet-Rich Plasma / cytology*
  • Platelet-Rich Plasma / metabolism
  • Receptors, Thrombin / chemistry
  • Thrombin / pharmacology


  • Annexin A5
  • P-Selectin
  • Receptors, Thrombin
  • Epidermal Growth Factor
  • Octoxynol
  • Thrombin