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Safety and bactericidal efficacy of cold atmospheric plasma generated by a flexible surface Dielectric Barrier Discharge device against Pseudomonas aeruginosa in vitro and in vivo.
Annals of Clinical Microbiology and Antimicrobials ( IF 5.7 ) Pub Date : 2020-08-19 , DOI: 10.1186/s12941-020-00381-z
Gabrielle S Dijksteel 1, 2 , Magda M W Ulrich 1, 2, 3 , Marcel Vlig 1 , Ana Sobota 4 , Esther Middelkoop 1, 2 , Bouke K H L Boekema 1
Affiliation  

Cold atmospheric plasma (CAP), which is ionized gas produced at atmospheric pressure, could be a novel and potent antimicrobial therapy for the treatment of infected wounds. Previously we have shown that CAP generated with a flexible surface Dielectric Barrier Discharge (sDBD) is highly effective against bacteria in vitro and in ex vivo burn wound models. In the current paper, we determined the in vitro and in vivo safety and efficacy of CAP generated by this sDBD device. The effect of CAP on DNA mutations of V79 fibroblasts was measured using a hypoxanthine–guanine-phosphoribosyltransferase (HPRT) assay. Furthermore, effects on cell proliferation, apoptosis and DNA damage in ex vivo burn wound models (BWMs) were assessed using immunohistochemistry. Next, 105 colony forming units (CFU) P. aeruginosa strain PAO1 were exposed to CAP in a 3D collagen-elastin matrix environment to determine the number of surviving bacteria in vitro. Finally, rat excision wounds were inoculated with 107 CFU PAO1 for 24 h. The wounds received a single CAP treatment, repeated treatments on 4 consecutive days with CAP, 100 µL of 1% (wt/wt) silver sulfadiazine or no treatment. Wound swabs and punch biopsies were taken to determine the number of surviving bacteria. Exposure of V79 fibroblasts to CAP did not increase the numbers of mutated colonies. Additionally, the number of proliferative, apoptotic and DNA damaged cells in the BWMs was comparable to that of the unexposed control. Exposure of PAO1 to CAP for 2 min resulted in the complete elimination of bacteria in vitro. Contrarily, CAP treatment for 6 min of rat wounds colonized with PAO1 did not effectively reduce the in vivo bacterial count. CAP treatment was safe but showed limited efficacy against PAO1 in our rat wound infection model.

中文翻译:

柔性表面介电屏障放电装置在体外和体内对铜绿假单胞菌产生的冷大气等离子体的安全性和杀菌功效。

冷大气等离子体(CAP)是在大气压下产生的离子化气体,可能是一种新颖有效的抗菌疗法,可用于治疗感染的伤口。以前我们已经表明,用柔性表面介电势垒放电(sDBD)生成的CAP在体外和离体烧伤创面模型中对细菌具有很高的抵抗力。在本文中,我们确定了此sDBD设备产生的CAP的体外和体内安全性和有效性。使用次黄嘌呤-鸟嘌呤-磷酸核糖基转移酶(HPRT)测定法测量了CAP对V79成纤维细胞DNA突变的影响。此外,使用免疫组织化学评估了离体烧伤创面模型(BWM)对细胞增殖,凋亡和DNA损伤的影响。接下来,有105个菌落形成单位(CFU)P。将铜绿假单胞菌菌株PAO1在3D胶原蛋白-弹性蛋白基质环境中暴露于CAP,以确定体外存活细菌的数量。最后,用107 CFU PAO1接种大鼠切除伤口24小时。伤口接受单次CAP治疗,连续4天用CAP,100 µL 1%(wt / wt)磺胺嘧啶银进行重复治疗或不进行治疗。取伤口拭子和打孔活组织检查以确定存活细菌的数量。V79成纤维细胞暴露于CAP并没有增加突变菌落的数量。此外,BWMs中增殖,凋亡和DNA损伤的细胞数量与未暴露的对照组相当。PAO1暴露于CAP 2分钟导致了体外细菌的完全清除。相反,CAP处理定植于PAO1的大鼠伤口6分钟并不能有效减少体内细菌数量。在我们的大鼠伤口感染模型中,CAP治疗是安全的,但对PAO1的疗效有限。
更新日期:2020-08-19
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