Saulis Gintautas, Saulė Rita
Laboratory of Biophysics for Bionanotechnology and Medicine, Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, 58 K. Donelaicio str., Kaunas, LT-44248, Lithuania.
Biochim Biophys Acta. 2012 Dec;1818(12):3032-9. doi: 10.1016/j.bbamem.2012.06.018. Epub 2012 Jul 2.
Here, the sizes of the pores created by square-wave electric pulses with the duration of 100 μs and 2 ms are compared for pulses with the amplitudes close to the threshold of electroporation. Experiments were carried out with three types of cells: mouse hepatoma MH-22A cells, Chinese hamster ovary (CHO) cells, and human erythrocytes. In the case of a short pulse (square-wave with the duration of 100 μs or exponential with the time constant of 22 μs), in the large portion (30-60%) of electroporated (permeable to potassium ions) cells, an electric pulse created only the pores, which were smaller than the molecule of bleomycin (molecular mass of 1450 Da, r≈0.8 nm) or sucrose (molecular mass of 342.3 Da, radius-0.44-0.52 nm). In the case of a long 2-ms duration pulse, in almost all cells, which were electroporated, there were the pores larger than the molecules of bleomycin and/or sucrose. Kinetics of pore resealing depended on the pulse duration and was faster after the shorter pulse. After a short 100-μs duration pulse, the disappearance of the pores permeable to bleomycin was completed after 6-7 min at 24-26°C, while after a long 2-ms duration pulse, this process was slower and lasted 15-20 min. Thus, it can be concluded that a short 100-μs duration pulse created smaller pores than the longer 2-ms duration pulse. This could be attributed to the time inadequacy for pores to grow and expand during the pulse, in the case of short pulses.
在此,对持续时间为100 μs和2 ms的方波电脉冲在接近电穿孔阈值的幅度下所产生的孔的大小进行了比较。实验使用了三种类型的细胞:小鼠肝癌MH - 22A细胞、中国仓鼠卵巢(CHO)细胞和人类红细胞。在短脉冲(持续时间为100 μs的方波或时间常数为22 μs的指数脉冲)的情况下,在大部分(30 - 60%)电穿孔(对钾离子通透)的细胞中,电脉冲仅产生了小于博来霉素分子(分子量1450 Da,r≈0.8 nm)或蔗糖分子(分子量342.3 Da,半径 - 0.44 - 0.52 nm)的孔。在持续时间为2 ms的长脉冲情况下,在几乎所有电穿孔的细胞中,都存在大于博来霉素和/或蔗糖分子的孔。孔重新封闭的动力学取决于脉冲持续时间,短脉冲后更快。在持续时间为100 μs的短脉冲后,在24 - 26°C下,对博来霉素通透的孔在6 - 7分钟后消失,而在持续时间为2 ms的长脉冲后,这个过程较慢,持续15 - 20分钟。因此,可以得出结论,持续时间为100 μs的短脉冲产生的孔比持续时间为2 ms的长脉冲产生的孔小。这可能归因于在短脉冲情况下,脉冲期间孔生长和扩张的时间不足。