Nanosecond pulsed electric field (nsPEF) has the ability to induce a host of intracellular biochemical processes in living cells as a function of parameter setting. In vitro experiments proved that nsPEF stimulation could remarkably promote biomineralization processes and new bone formation. However, the impact of nsPEF parameter settings on the calcium flux of osteoblasts, as well as the intracellular mechanisms underlying those observations have yet to be elucidated. In this study, live osteoblast-like MG63 cells were loaded with fluorescence indicator dye fluo-4 AM. nsPEF stimulation could induce intracellular calcium mobilization in MG63 cells with no refractory period. We confirmed that moderate output voltage (4–8 KV) and large pulse number (25) were required for efficient nsPEF irritation of MG63 cells. Additionally, nsPEF stimulation-induced calcium flux in MG63 cells was dramatically reduced with the treatment by ethylene glycol tetraacetic acid (EGTA), proving that the intake of extracellular calcium ions is crucial for increasing intracellular calcium concentration of nsPEF-treated MG63 cells. Our preliminary study investigated the mechanisms underlying nsPEF stimulation-induced calcium mobilization in osteoblast-like cells, and it has the potential to accelerate the application of nsPEF stimulation in new bone formation.

纳秒脉冲电场（nsPEF）具有根据参数设置诱导活细胞中许多细胞内生化过程的能力。体外实验证明，nsPEF刺激可以显着促进生物矿化过程和新骨形成。但是，尚未阐明nsPEF参数设置对成骨细胞钙通量的影响以及这些观察所依据的细胞内机制。在这项研究中，活的成骨细胞样MG63细胞装有荧光指示剂fluo-4 AM。nsPEF刺激可以诱导MG63细胞中的细胞内钙动员，没有不应期。我们证实，有效的nsPEF刺激MG63细胞需要适度的输出电压（4-8 KV）和大脉冲数（25）。此外，乙二醇四乙酸（EGTA）的处理大大降低了nsPEF刺激诱导的MG63细胞的钙通量，证明摄入细胞外钙离子对于增加nsPEF处理的MG63细胞的细胞内钙浓度至关重要。我们的初步研究调查了nsPEF刺激诱导的成骨样细胞钙动员的机制，它具有加速nsPEF刺激在新骨形成中的应用的潜力。