高压纳秒脉冲电场消融黑色素瘤细胞实验研究

Experimental study on melanoma cell ablation by high-voltagenanosecond pulsed electric field

为了研究高压纳秒脉冲电场(nsPEF)消融恶性肿瘤的关键影响参数,基于火花开关和传输线变压器(TLT)技术,自主研制重频高压纳秒脉冲电场(RnsPEF)发生系统,可以稳定输出纳秒级脉宽的指数脉冲,证实了高压纳秒脉冲电场杀伤肿瘤细胞的效果和可控性.以贴壁生长于六孔板中B16黑色素瘤细胞为对象,研究脉冲次数、峰值电压、重复频率和电极针对间距对肿瘤细胞消融效果的影响.以电极杯中B16肿瘤细胞悬液为研究对象,结合CCK-8检测法开展脉冲处理后细胞活性的研究.结果发现,高压脉冲电场和脉冲能量注入密度是影响纳秒脉冲电场消融肿瘤细胞的关键因素,重复频率对消融效果的影响不大.结果显示,自制RnsPEF系统消融B16肿瘤细胞的阈值电场强度为6.8 kV/cm,注入能量密度的阈值为11.4 J/cm3和最佳消融次数为500次脉冲.

A repetitive high-voltage nanosecond pulsed electric field(RnsPEF)generation system was independently developed based on the spark switch and transmission line transformer(TLT)technology in order to analyze the key impact parameters of the process of malignant tumors ablation by high-voltage nanosecond pulsed electric field(nsPEF).The system can stably generate nanosecond exponential pulse.The experimental results proved the effectivity and controllability of RnsPEF on tumor cells ablation.B16 melanoma cells adherently seeded in six-well plates as the research object to analyze the effects of pulse number,peak voltage,repetition frequency and electrode spacing on tumor cells ablation.Cell counting kit-8(CCK-8)was applied to measure cell viability of B16 tumor cells suspension in the cuvette after treated by pulses.The experimental results show that the pulsed electric field intensity and injected energy density of the applied RnsPEF play the key roles in determining the ablation effect.The repetition frequency hardly affects the ablation results.The pulsed electric field intensity threshold of RnsPEF ablating B16 melanoma cells is 6.8 kV/cm,and the injected energy density threshold is 11.4 J/cm3,as well as the optimal pulse number is 500 pulses.