A multi-donor ex vivo platelet activation and growth factor release study using electric pulses with durations up to 100 microseconds

Abstract

In vitro platelet activation is established in various clinical settings for wound healing and tissue regeneration. After separating platelet rich plasma (PRP) from whole blood, a biochemical activator, typically bovine thrombin (BT), is applied to activate PRP, which clots the PRP and releases growth factors beneficial for wound healing. BT's drawbacks, particularly cost, availability, workflow challenges, and potential immune responses, motivated the development of ex vivo electrical activation of PRP using pulsed electric fields (PEFs) with durations on the order of hundreds of nanoseconds and electric field strengths from ∼10-100 kV/cm. PEFs permeabilize platelets to facilitate Ca2+ transport into the cells to induce platelet activation; however, membrane permeabilization does not require PEFs of such short duration and high intensity. This study demonstrates that 5-100 µs duration PEFs effectively activate platelets. Treating PRP with ten different single pulse waveforms with durations between 5 and 100 µs induces similar or higher levels of platelet derived, vascular endothelial, and epidermal growth factor release compared to BT. These results indicate the potential clinical relevance of microsecond PEFs for platelet stimulation, which may reduce the expense and device footprint for PEF mediated platelet stimulation compared to nanosecond pulse generators, facilitating technology transition for clinical and trauma applications.