Khan Saiqa I, Blumrosen Gaddi, Vecchio Daniela, Golberg Alexander, McCormack Michael C, Yarmush Martin L, Hamblin Michael R, Austen William G
Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114.
Department of Computer Science, Tel Aviv University, Tel Aviv, Israel.
Biotechnol Bioeng. 2016 Mar;113(3):643-650. doi: 10.1002/bit.25818. Epub 2015 Sep 9.
Biofilm formation is a significant problem, accounting for over eighty percent of microbial infections in the body. Biofilm eradication is problematic due to increased resistance to antibiotics and antimicrobials as compared to planktonic cells. The purpose of this study was to investigate the effect of Pulsed Electric Fields (PEF) on biofilm-infected mesh. Prolene mesh was infected with bioluminescent Pseudomonas aeruginosa and treated with PEF using a concentric electrode system to derive, in a single experiment, the critical electric field strength needed to kill bacteria. The effect of the electric field strength and the number of pulses (with a fixed pulse length duration and frequency) on bacterial eradication was investigated. For all experiments, biofilm formation and disruption were confirmed with bioluminescent imaging and Scanning Electron Microscopy (SEM). Computation and statistical methods were used to analyze treatment efficiency and to compare it to existing theoretical models. In all experiments 1500 V are applied through a central electrode, with pulse duration of 50 μs, and pulse delivery frequency of 2 Hz. We found that the critical electric field strength (Ecr) needed to eradicate 100-80% of bacteria in the treated area was 121 ± 14 V/mm when 300 pulses were applied, and 235 ± 6.1 V/mm when 150 pulses were applied. The area at which 100-80% of bacteria were eradicated was 50.5 ± 9.9 mm(2) for 300 pulses, and 13.4 ± 0.65 mm(2) for 150 pulses. 80% threshold eradication was not achieved with 100 pulses. The results indicate that increased efficacy of treatment is due to increased number of pulses delivered. In addition, we that showed the bacterial death rate as a function of the electrical field follows the statistical Weibull model for 150 and 300 pulses. We hypothesize that in the clinical setting, combining systemic antibacterial therapy with PEF will yield a synergistic effect leading to improved eradication of mesh infections.
生物膜形成是一个重大问题,占人体微生物感染的80%以上。与浮游细胞相比,生物膜对抗生素和抗菌剂的耐药性增加,因此根除生物膜存在问题。本研究的目的是调查脉冲电场(PEF)对生物膜感染的网片的影响。将普理灵网片感染生物发光的铜绿假单胞菌,并使用同心电极系统对其进行PEF处理,以便在单一实验中得出杀死细菌所需的临界电场强度。研究了电场强度和脉冲数(在固定脉冲长度持续时间和频率的情况下)对细菌根除的影响。对于所有实验,通过生物发光成像和扫描电子显微镜(SEM)确认生物膜的形成和破坏情况。使用计算和统计方法分析治疗效率,并将其与现有的理论模型进行比较。在所有实验中,通过中心电极施加1500V电压,脉冲持续时间为50μs,脉冲传递频率为2Hz。我们发现,当施加300个脉冲时,在治疗区域根除100%-80%细菌所需的临界电场强度(Ecr)为121±14V/mm;当施加150个脉冲时,该强度为235±6.1V/mm。对于300个脉冲,根除100%-80%细菌的区域为50.5±9.9mm²;对于150个脉冲,该区域为13.4±0.65mm²。施加100个脉冲未达到80%的根除阈值。结果表明,治疗效果的提高是由于施加的脉冲数增加。此外,我们发现对于150个和300个脉冲,细菌死亡率与电场的函数关系遵循统计韦布尔模型。我们假设在临床环境中,将全身抗菌治疗与PEF相结合将产生协同效应,从而改善网片感染的根除效果。