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 $\sim 10$ -100 kV/cm. PEFs permeabilize platelets to facilitate Ca ²⁺ 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- $100~\mu \text{s}$ duration PEFs effectively activate platelets. Treating PRP with ten different single pulse waveforms with durations between 5 and $100~\mu \text{s}$ 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.

中文翻译：

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使用持续时间长达100微秒的电脉冲进行多供体离体血小板活化和生长因子释放研究

在各种临床环境中建立了用于伤口愈合和组织再生的体外血小板活化。从全血中分离出富含血小板的血浆（PRP）之后，将生化活化剂（通常是牛凝血酶（BT））用于活化PRP，这会凝结PRP并释放出有利于伤口愈合的生长因子。BT的缺点（尤其是成本，可用性，工作流程挑战和潜在的免疫反应）的缺点促使人们开发使用脉冲电场（PEF）进行PRP的离体电激活，持续时间约为数百纳秒，电场强度为 $ \ sim 10 $ -100 kV / cm。PEFs透化血小板以促进Ca ²⁺转运进入细胞以诱导血小板活化。然而，膜通透性不需要持续时间短且强度高的PEF。这项研究表明，5- $ 100〜\ mu \ text {s} $ 持续时间的PEF有效激活血小板。用10种不同的单脉冲波形治疗PRP，持续时间在5到5之间 $ 100〜\ mu \ text {s} $ 与BT相比，可诱导相似或更高水平的血小板衍生，血管内皮和表皮生长因子释放。这些结果表明微秒PEF对血小板刺激的潜在临床相关性，与纳秒脉冲发生器相比，可以减少PEF介导的血小板刺激的费用和设备占用空间，从而促进了临床和创伤应用的技术过渡。