Tunable activation of therapeutic platelet-rich plasma by pulse electric field: Differential effects on clot formation, growth factor release, and platelet morphology

• Published in: PloS one 2018-09, Vol.13 (9), p.e0203557-e0203557
• Main Authors: Frelinger, 3rd, Andrew L, Gerrits, Anja J, Neculaes, V Bogdan, Gremmel, Thomas, Torres, Andrew S, Caiafa, Anthony, Carmichael, Sabrina L, Michelson, Alan D
• Format: Article
• Language: English
• Subjects: Activation; Analysis; Angiogenesis; Biology and Life Sciences; Blood; Blood diseases; Blood platelets; Calcium chloride; Cancer; Dielectric properties; Electric fields; Electrochemical Techniques; Electron microscopy; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor - metabolism; Flow cytometry; Gels; Granular materials; Granulation; Growth factors; Healthy Volunteers; Humans; Intercellular Signaling Peptides and Proteins - metabolism; Laboratories; Markers; Mechanical properties; Medical schools; Medicine and Health Sciences; Microscopy, Electron, Transmission; Modulus of elasticity; Morphology; P-selectin; P-Selectin - metabolism; Platelet Activation - physiology; Platelet-derived growth factor; Platelet-Derived Growth Factor - metabolism; Platelet-rich plasma; Platelet-Rich Plasma - metabolism; Platelets; Polarity; Protein disulfide-isomerase; Proteins; Research and Analysis Methods; Sports medicine; Studies; Thrombelastography; Thrombin; Tissue engineering; TLR9 protein; Toll-Like Receptor 9 - metabolism; Toll-like receptors; Transmission electron microscopy; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; Wound care; Wound healing
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0203557

Activation of platelet-rich plasma (PRP) by pulse electric field (PEF) releases growth factors which promote wound healing (e.g., PDGF, VEGF for granulation, EGF for epithelialization). To determine after PEF activation of therapeutic PRP: 1) platelet gel strength; 2) profile of released growth factors; 3) alpha- and T-granule release; 4) platelet morphology. Concentrated normal donor PRP was activated by 5 μsec (long) monopolar pulse, ~4000 V/cm (PEF A) or 150 nsec (short) bipolar pulse, ~3000 V/cm (PEF B) in the presence of 2.5 mM (low) or 20 mM (high) added CaCl2. Clot formation was evaluated by thromboelastography (TEG). Surface exposure of alpha granule (P-selectin) and T-granule (TLR9 and protein disulfide isomerase [PDI]) markers were assessed by flow cytometry. Factors in supernatants of activated PRP were measured by ELISA. Platelet morphology was evaluated by transmission electron microscopy (TEM). Time to initial clot formation was shorter with thrombin (