[利用冷大气等离子体灭活念珠菌在根除表面真菌感染的新方法中]。

Ewa Tyczkowska-Sieroń, Justyna Markiewicz
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引用次数: 0

摘要

近年来,医院真菌感染已成为日益严重的问题。念珠菌在皮肤和粘膜上的定植在高危人群中引起的并发症越来越多。因此,快速有效地治疗念珠菌引起的浅表真菌感染是现代医学的一项重要任务。不幸的是,随着真菌感染数量的明显增加,目前使用的抗真菌药物的耐药性也在增加,严重限制了治疗的有效性。因此,有必要深入研究新的治疗方法。一个很有前途的解决方案是使用低温大气等离子体。本文的目的是探讨这类血浆对白色念珠菌存活的影响。方法:采用一种称为等离子剃刀的线性微放电射流作为低温大气等离子体源。等离子体在13.56 MHz下产生,使用He作为反应气体。气体流量为1.9 L/min,放电功率为17 W。实验系统的示意图如图1所示。以参考菌株白色念珠菌ATCC 10231为模型材料进行研究。将含5 × 10(7)个细胞/mL的100 μL磷酸盐缓冲盐水均匀涂布在培养皿表面制备培养物。这种培养物在不同时间暴露在血浆中。通过密度法估计真菌生长抑制区的大小(图3)。为了获得更完整的等离子体信息,测量了光学发射光谱。结果:发现随着血浆治疗时间的延长,抑制区明显增大(图2)。图4为抑制区大小随治疗时间变化的实验结果。根据公式(5)绘制的理论曲线(图4)成功地拟合了这些结果(p = 0.0058, r2 = 0.944),该曲线是基于等离子体中心杀伤剂扩散的简单模型得出的。光学发射光谱的研究证实了冷大气等离子体中可能产生的各种杀伤剂,如紫外线、自由基、离子和高能电子。进一步的研究将集中在确定负责细胞杀伤过程的主要药物,并确定这一过程的机制。结论:等离子剃刀产生的低温大气等离子体是一种非常有效的杀灭病原菌的工具。虽然目前的研究只是关于低温大气微等离子体对真菌细胞影响的初步研究,但它们为使用这种技术作为根除表面真菌感染的方法提供了希望。
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[Inactivation of Candida species using cold atmospheric plasma on the way to a new method of eradication of superficial fungal infections].

Introduction: In recent years, nosocomial fungal infections are becoming an increasingly serious problem. More and more complications are observed in patients with high-risk groups resulting from the colonization of the skin and mucous membranes by Candida species. Thus, rapid and effective treatment of superficial fungal infections caused by Candida is a very important task for modern medicine. Unfortunately, with a clear increase in the number of fungal infections, the resistance to currently used antifungal drugs also increases seriously limited the effectiveness of treatment. An intensive search for new therapeutic solutions is therefore necessary. One of the promising solutions is the use of cold atmospheric plasma. The aim of this paper is to investigate the influence of this type of plasma on survival of Candida albicans.

Methods: As a source of cold atmospheric plasma, a linear microdischarge jet, called plasma razor, was used. Plasma was generated at 13.56 MHz, using He as a reactive gas. The gas flow rate and the discharge power were 1.9 L/min and 17 W, respectively. A schematic view of the experimental system is shown in Fig. 1. The reference strain of Candida albicans ATCC 10231 was used as a model material for investigations. The culture was prepared by spreading uniformly 100 μL phosphate buffered saline solution containing 5 x10(7) cells/mL on the surface of a Petri dish. Such a culture was exposed to the plasma at various times. The size of the zone of inhibition of fungal growth was estimated by densitometric method (Fig. 3). For more complete information about the plasma the optical emission spectra were measured.

Results: It was found that with increasing time of plasma treatment, the zone of inhibition clearly increases (Fig. 2). In Fig. 4, the experimental results of the size of the inhibition zone versus the treatment time are shown. These results were successfully fitted (p = 0.0058, r2 = 0.944) by a theoretical curve (Fig. 4), plotted according to Eq. (5), which was derived on the basis of a simple model of the spread of a killing agent from the plasma center. The study of the optical emission spectra confirmed a large variety of possible killing agents generated in the cold atmospheric plasma, such as UV, radicals, ions and energetic electrons. Further research will be focused on the determination of the main agent responsible for the process of the cell killing, and to determine the mechanism of this process.

Conclusions: Cold atmospheric plasma generated by the plasma razor turns out to be a very effective tool for the killing of pathogenic fungi. Although the presented studies are only the initial stage of work on the effects of cold atmospheric microplasma on fungal cells, they provide hope for the possibility of using this technique as a method of eradication of superficial fungal infections.

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