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Selective Inactivation of Pseudomonas aeruginosa and Staphylococcus epidermidis with Pulsed Electric Fields and Antibiotics.

ABSTRACT: Objective: Increasing numbers of multidrug-resistant bacteria make many antibiotics ineffective; therefore, new approaches to combat microbial infections are needed. In addition, antibiotics are not selective-they kill pathogenic organisms as well as organisms that could positively contribute to wound healing (bio flora). Approach: Here we report on selective inactivation of Pseudomonas aeruginosa and Staphylococcus epidermidis, potential pathogens involved in wound infections with pulsed electric fields (PEFs) and antibiotics (mix of penicillin, streptomycin, and nystatin). Results: Using a Taguchi experimental design in vitro, we found that, under similar electric field strengths, the pulse duration is the most important parameter for P. aeruginosa inactivation, followed by the number of pulses and pulse frequency. P. aeruginosa, a potential severe pathogen, is more sensitive than the less pathogenic S. epidermidis to PEF (alone or in combination with antibiotics). Applying 200 pulses with a duration of 60??s at 2.8?Hz, the minimum electric fields of 308.8?±?28.3 and 378.4?±?12.9?V/mm were required to inactive P. aeruginosa and S. epidermidis, respectively. Addition of antibiotics reduced the threshold for minimum electric fields required to inactivate the bacteria. Innovation: This study provides essential information, such as critical electric field parameters for bacteria inactivation, required for developing in vivo treatment and clinical protocols for using PEF for wound healing. Conclusion: A combination of PEFs with antibiotics reduces the electric field threshold required for bacteria disinfection. Such an approach simplifies devices required to disinfect large areas of infected wounds.

SUBMITTER: Rubin AE

PROVIDER: S-EPMC6855282 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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Selective Inactivation of Pseudomonas aeruginosa and Staphylococcus epidermidis with Pulsed Electric Fields and Antibiotics.

Rubin Andrey Ethan AE Usta Osman Berk OB Schloss Rene R Yarmush Martin M Golberg Alexander A

Advances in wound care 20190403 4

Objective: Increasing numbers of multidrug-resistant bacteria make many antibiotics ineffective; therefore, new approaches to combat microbial infections are needed. In addition, antibiotics are not selective-they kill pathogenic organisms as well as organisms that could positively contribute to wound healing (bio flora). Approach: Here we report on selective inactivation of Pseudomonas aeruginosa and Staphylococcus epidermidis, potential pathogens involved in wound i ...[more]

PMID: 31737412

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Project description:Staphylococcus epidermidis is implicated in a multitude of human infections and is one of the major causes of clinical infections in hospitals, especially at surgical sites and on indwelling medical devices, such as orthopedic implants. These infections are especially dangerous because of the S. epidermidis propensity to form biofilms, which increases resistance to antibiotics and the natural immune response. This study investigated pulsed electromagnetic fields (PEMF) as a potential treatment to combat such infections, as PEMF exposure was expected to disrupt the electrostatic forces that adhere staphylococcal cells to surfaces and to one another. To test the effect of PEMF on biofilms, S. epidermidis cultures were exposed to PEMF at various durations either during the growth phase or after a full biofilm had formed. In addition, cells were exposed to PEMF and concomitant antibiotic treatment. Biofilm viability was quantified by both crystal violet and alamarBlue assays and scanning electron microscopy. The results demonstrated that PEMF significantly inhibited biofilm formation and disrupted preformed biofilms in vitro while also showing synergistic biofilm inhibition when combined with antibiotics. These combined results indicate that PEMF should be considered a promising novel technique for treating S. epidermidis biofilm infections and undergo further testing in vivo. IMPORTANCE Antibiotic resistance and biofilm infections are major issues in health care because of the lack of a successful treatment modality and poor patient outcomes. These infections are a particular issue following orthopedic surgery or trauma wherein an infection may form on an orthopedic implant or patient's bone. The presented study demonstrates that pulsed electromagnetic fields may be a promising novel treatment for such infections and can overcome the medical challenges presented by biofilm formation. Furthermore, the effects demonstrated are even greater when combining pulsed electromagnetic field therapy with traditional antibiotics.

Project description:Objective: The incidence of severe infectious complications after burn injury increases mortality by 40%. However, traditional approaches for managing burn infections are not always effective. High-voltage, pulsed electric field (PEF) treatment shortly after a burn injury has demonstrated an antimicrobial effect in vivo; however, the working parameters and long-term effects of PEF treatment have not yet been investigated. Approach: Nine sets of PEF parameters were investigated to optimize the applied voltage, pulse duration, and frequency or pulse repetition for disinfection of Pseudomonas aeruginosa infection in a stable mouse burn wound model. The bacterial load after PEF administration was monitored for 3 days through bioluminescence imaging. Histological assessments and inflammation response analyses were performed at 1 and 24 h after the therapy. Results: Among all tested PEF parameters, the best disinfection efficacy of P. aeruginosa infection was achieved with a combination of 500 V, 100 μs, and 200 pulses delivered at 3 Hz through two plate electrodes positioned 1 mm apart for up to 3 days after the injury. Histological examinations revealed fewer inflammatory signs in PEF-treated wounds compared with untreated infected burns. Moreover, the expression levels of multiple inflammatory-related cytokines (interleukin [IL]-1α/β, IL-6, IL-10, leukemia inhibitory factor [LIF], and tumor necrosis factor-alpha [TNF-α]), chemokines (macrophage inflammatory protein [MIP]-1α/β and monocyte chemoattractant protein-1 [MCP-1]), and inflammation-related factors (vascular endothelial growth factor [VEGF], macrophage colony-stimulating factor [M-CSF], and granulocyte-macrophage colony-stimulating factor [G-CSF]) were significantly decreased in the infected burn wound after PEF treatment. Innovation: We showed that PEF treatment on infected wounds reduces the P. aeruginosa load and modulates inflammatory responses. Conclusion: The data presented in this study suggest that PEF treatment is a potent candidate for antimicrobial therapy for P. aeruginosa burn infections.

Project description:Glioblastoma multiforme (GBM), is the most common form of adult malignant brain tumor, highly heterogeneous and relatively resistant to therapy with a poor prognosis. As other cancers, GBM starts from a relatively small fraction of poorly differentiated and particularly aggressive cancer stem cells (CSCs), responsible for aberrant proliferation and invasion, rapidly spreading the tumor growth in the other parts of the central nervous system. Due to the extreme tumor heterogeneity, standard therapies (i.e., surgery, chemotherapy and radiotherapy) provide poor positive outcomes and cancers usually recur. Therefore, alternative approaches, possibly targeting CSCs, are necessary for searching effective therapeutic strategies against GBM. Among many emerging new therapeutic strategies, ultra-short pulsed electric fields (PEF) are considered extremely promising in cancer therapy and our previous results demonstrated the capability of a specific electric pulse protocol, characterized by a high pulse amplitude and a short duration, to selectively affect medulloblastoma CSCs preserving normal cells. Here, we tested the same exposure protocol to investigate the response of U87 GBM cells, and of U87-derived neurospheres. By analyzing different in vitro biological endpoints and taking advantage of transcriptomic and bioinformatics analyses, we found that, independently of CSCs content, PEF exposure affected cell proliferation and differentially regulated hypoxia, inflammation and P53/cell cycle checkpoints. PEF exposure also significantly reduced the clonogenic potential, reducing the ability to form new neurospheres, and inhibited the invasion potential. Importantly, exclusively in U87 neurospheres, PEF exposure changed the expression of stemness/differentiation genes. Our results, confirm this physical stimulus as a promising treatment to destabilize the extreme intra-tumoral GBM heterogeneity opening up the possibility to develop future therapeutic strategies mediated by PEF exposure.

Dataset Information

Selective Inactivation of Pseudomonas aeruginosa and Staphylococcus epidermidis with Pulsed Electric Fields and Antibiotics.

Publications

Selective Inactivation of <i>Pseudomonas aeruginosa</i> and <i>Staphylococcus epidermidis</i> with Pulsed Electric Fields and Antibiotics.

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