title: Platelet-rich plasma stimulated by pulse electric fields: Platelet activation, procoagulant markers, growth factor release and cell proliferation.
abstract: 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.
authors.affiliation: a Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School , Boston , MA , USA and.
authors.forename: A L
authors.initials: AL
authors.lastname: Frelinger
authors.affiliation: b GE Global Research Center , Niskayuna , NY , USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Torres
authors.affiliation: b GE Global Research Center , Niskayuna , NY , USA.
authors.forename: A
authors.initials: A
authors.lastname: Caiafa
authors.affiliation: b GE Global Research Center , Niskayuna , NY , USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Morton
authors.affiliation: a Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School , Boston , MA , USA and.
authors.forename: M A
authors.initials: MA
authors.lastname: Berny-Lang
authors.affiliation: a Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School , Boston , MA , USA and.
authors.forename: A J
authors.initials: AJ
authors.lastname: Gerrits
authors.affiliation: a Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School , Boston , MA , USA and.
authors.forename: S L
authors.initials: SL
authors.lastname: Carmichael
authors.affiliation: b GE Global Research Center , Niskayuna , NY , USA.
authors.forename: V B
authors.initials: VB
authors.lastname: Neculaes
authors.affiliation: a Center for Platelet Research Studies, Division of Hematology/Oncology, Boston Children's Hospital, Dana-Farber Cancer Institute, Harvard Medical School , Boston , MA , USA and.
authors.forename: A D
authors.initials: AD
authors.lastname: Michelson
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000096:biosynthesis / Q000235:genetics; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D055252:Cell-Derived Microparticles / Q000378:metabolism; D004558:Electric Stimulation; D004560:Electricity; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D017830:Octoxynol / Q000494:pharmacology; D019007:P-Selectin / Q000096:biosynthesis / Q000235:genetics; D015539:Platelet Activation / Q000187:drug effects; D053657:Platelet-Rich Plasma / Q000166:cytology* / Q000378:metabolism; D018179:Receptors, Thrombin / Q000737:chemistry; D013917:Thrombin / Q000494:pharmacology
chemical_list: D017304:Annexin A5; D019007:P-Selectin; D018179:Receptors, Thrombin; D004815:Epidermal Growth Factor; D017830:Octoxynol; D013917:Thrombin
keywords: Growth factors; platelet-rich plasma; platelets; pulse electric field; wound healing