Pessoa Palloma Santiago Prates, Barcelos Raphael, Pinto Larissa Fagundes, de Freitas Denise, Campos Mauro
Department of Ophthalmology and Visual Sciences, Paulista School of Medicine, São Paulo Hospital, Federal University of São Paulo, São Paulo, São Paulo, Brazil - UNIFESP.
PLoS One. 2025 Feb 25;20(2):e0317409. doi: 10.1371/journal.pone.0317409. eCollection 2025.
Species of the genus Acanthamoeba spp. are ubiquitous and can cause Acanthamoeba keratitis (AK), a serious corneal infection. Due to the toxicity and ineffectiveness of currently available prolonged therapies, we investigated electroceutical treatment aimed at facilitating the permeation of molecules through the membrane of cysts and trophozoites, which allows for faster elimination of the parasite.
Cysts and trophozoites of Acanthamoeba polyphaga (ATCC® 30461TM) were exposed in vitro to an electric field with intensities of 2,000 volts and 2,500 volts. Viability after electroporation was assessed by the exclusion method with 0.4% trypan blue dye, while permeabilization was assessed by fluorescence microscopy using propidium iodide (PI), since both are impermeable to the membrane of viable and intact cells. The images were acquired on a Nikon Eclipse TI-U microscope and analyzed using ImageJ software.
With regard to viability, 40% of the trophozoites electroporated at 2,000 V and 42% of those electroporated at 2,500 V were lost, while for cysts the loss was 13% and 16% respectively. Considering permeabilization, 55% of trophozoites and cysts were permeabilized at 2,000 V (p ≤ 0.05); and 59% at 2,500 V for both (p ≤ 0.05). Values of p < 0.05 were considered statistically significant.
The voltages tested were effective for both cysts and trophozoites, since the percentages of permeabilization were close, with no statistical significance between them, only with the control groups. These results suggest the possibility that an electroceutical treatment could be applied as a complement to the standard treatment for AK.
棘阿米巴属物种广泛存在，可引起棘阿米巴角膜炎（AK），这是一种严重的角膜感染。由于目前可用的长期治疗方法存在毒性且效果不佳，我们研究了电治疗方法，旨在促进分子透过包囊和滋养体的膜，从而更快地清除寄生虫。
将多食棘阿米巴（ATCC® 30461TM）的包囊和滋养体在体外暴露于强度为2000伏和2500伏的电场中。通过0.4%台盼蓝染料排除法评估电穿孔后的活力，同时使用碘化丙啶（PI）通过荧光显微镜评估通透性，因为这两种物质都不能透过存活且完整的细胞膜。图像在尼康Eclipse TI-U显微镜上采集，并使用ImageJ软件进行分析。
关于活力，在2000伏电穿孔的滋养体中有40%丧失活力，在2500伏电穿孔的滋养体中有42%丧失活力，而包囊的丧失率分别为13%和16%。考虑通透性，在2000伏时，55%的滋养体和包囊发生了通透性改变（p≤0.05）；在2500伏时，两者的通透性改变率均为59%（p≤0.05）。p<0.05的值被认为具有统计学意义。
所测试的电压对包囊和滋养体均有效，因为通透性改变的百分比相近，它们之间无统计学意义，仅与对照组有差异。这些结果表明电治疗方法有可能作为AK标准治疗的补充手段。
