Gallo S A, Sen A, Hensen M L, Hui S W
Membrane Biophysics Laboratory, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Biophys J. 1999 May;76(5):2824-32. doi: 10.1016/S0006-3495(99)77436-9.
The morphological changes to heat-stripped porcine stratum corneum following an electroporating pulse were studied by time-resolved freeze fracture electron microscopy. Pulses at a supra-electroporation threshold of 80 volts and 300 microseconds were applied across the stratum corneum with a pair of copper plate electrodes, which also served as cooling contacts. Multilamellar vesicles of 0.1-5.5 mm in diameter in the intercellular lipid bilayers of the stratum corneum appeared in less than milliseconds after pulsing. Pulsed samples exhibited aggregations of vesicles, whereas only occasional single vesicles were seen in the unpulsed samples. Aggregates form in less than a millisecond and disappear within minutes after the pulse. Their size ranged from 0.3 to 700 mm2. The size of individual vesicles, aggregate density, and size were analyzed as functions of postpulse time. These aggregate formations seem to be a secondary reaction to the pulse-induced skin permeabilization, determined by the resistance drop and recovery after the pulse. Heating the samples to 65 degrees C also caused vesicle aggregates of similar appearance to form, suggesting that these aggregations are related to the heating effect of the pulse. Hydration is thought to play an important role in aggregate formation.
通过时间分辨冷冻断裂电子显微镜研究了电穿孔脉冲后热剥离猪角质层的形态变化。使用一对铜板电极在角质层上施加80伏和300微秒的超电穿孔阈值脉冲,这对电极也用作冷却接触点。脉冲后不到一毫秒,角质层细胞间脂质双层中就出现了直径为0.1 - 5.5毫米的多层囊泡。脉冲处理的样品中囊泡出现聚集,而未脉冲处理的样品中仅偶尔可见单个囊泡。聚集体在不到一毫秒内形成,并在脉冲后几分钟内消失。它们的大小范围为0.3至700平方毫米。分析了单个囊泡的大小、聚集体密度和大小与脉冲后时间的关系。这些聚集体的形成似乎是对脉冲诱导的皮肤通透性改变的二次反应,由脉冲后的电阻下降和恢复决定。将样品加热到65摄氏度也会导致形成外观相似的囊泡聚集体,这表明这些聚集体与脉冲的热效应有关。水合作用被认为在聚集体形成中起重要作用。