Disinfection of ocular cells and tissues by atmospheric-pressure cold plasma.
Disinfection of ocular cells and tissues by atmospheric-pressure cold plasma.
复制标题
DOI：
10.1371/journal.pone.0033245
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Leonardi A
中科院分区：
文献类型：
--
作者：
Brun P;Brun P;Vono M;Venier P;Tarricone E;Deligianni V;Martines E;Zuin M;Spagnolo S;Cavazzana R;Cardin R;Castagliuolo I;Valerio AL;Leonardi A
低温等离子体已在医学上被提议作为组织消毒剂，并且由于细菌对抗生素频繁产生耐药性而受到越来越多的关注。这项研究探讨了一种电离氦气流的新型便携式设备产生的大气压冷等离子体（APCP）是否可以灭活眼部病原体而不造成明显的组织损伤。我们测试了 APCP 对培养的铜绿假单胞菌、大肠杆菌、金黄色葡萄球菌、白色念珠菌、烟曲霉和单纯疱疹病毒 1、眼细胞（结膜成纤维细胞和角膜细胞）和离体角膜的影响。暴露于 APCP 0.5 至 5 分钟可显着降低微生物活力（集落形成单位），但不会降低人类细胞活力（MTT 测定、FACS 和 Tunel 分析）或 HSV-1 空斑形成单位的数量。使用 FACS 激活的 2',7'-二氯荧光素二乙酸盐探针发现暴露的微生物和细胞中细胞内活性氧 (ROS) 水平增加。免疫测定表明在细胞培养物和角膜组织中没有诱导胸腺嘧啶二聚体。通过 HPLC、qRT-PCR 和蛋白质印迹分析，确定眼细胞和角膜中 8-OHdG、与氧化应激相关的基因和蛋白质（OGG1、GPX、NFE2L2）的表达短暂增加。尽管对结膜成纤维细胞和角膜细胞的治疗会导致细胞内ROS和氧化应激相关反应的时间限制性产生，但短期应用APCP似乎是一种有效且快速的细菌和真菌眼部消毒剂，不会对眼部细胞和组织造成明显损害。
Low temperature plasmas have been proposed in medicine as agents for tissue disinfection and have received increasing attention due to the frequency of bacterial resistance to antibiotics. This study explored whether atmospheric-pressure cold plasma (APCP) generated by a new portable device that ionizes a flow of helium gas can inactivate ocular pathogens without causing significant tissue damage. We tested the APCP effects on cultured Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Candida albicans, Aspergillus fumigatus and Herpes simplex virus-1, ocular cells (conjunctival fibroblasts and keratocytes) and ex-vivo corneas. Exposure to APCP for 0.5 to 5 minutes significantly reduced microbial viability (colony-forming units) but not human cell viability (MTT assay, FACS and Tunel analysis) or the number of HSV-1 plaque-forming units. Increased levels of intracellular reactive oxygen species (ROS) in exposed microorganisms and cells were found using a FACS-activated 2′,7′-dichlorofluorescein diacetate probe. Immunoassays demonstrated no induction of thymine dimers in cell cultures and corneal tissues. A transient increased expression of 8-OHdG, genes and proteins related to oxidative stress (OGG1, GPX, NFE2L2), was determined in ocular cells and corneas by HPLC, qRT-PCR and Western blot analysis. A short application of APCP appears to be an efficient and rapid ocular disinfectant for bacteria and fungi without significant damage on ocular cells and tissues, although the treatment of conjunctival fibroblasts and keratocytes caused a time-restricted generation of intracellular ROS and oxidative stress-related responses.
登录
查看更多内容
影响因子：
6.4
作者：
Lonkar, Pallavi;Dedon, Peter C.
通讯作者：
Dedon, Peter C.
影响因子：
3.3
作者：
Morfill, G. E.;Kong, M. G.;Zimmermann, J. L.
通讯作者：
Zimmermann, J. L.
影响因子：
4.4
作者：
Al-Adhami, B. H.;Nichols, R. A. B.;Smith, H. V.
通讯作者：
Smith, H. V.
影响因子：
3.1
作者：
CALLEGAN, MC;ENGEL, LS;OCALLAGHAN, RJ
通讯作者：
OCALLAGHAN, RJ
影响因子：
3.5
作者：
Fridman, Gregory;Brooks, Ari D.;Friedman, Gary
通讯作者：
Friedman, Gary