Bioelectric Effect Research Articles

The treatment of bacterial biofilms cultivated on knee arthroplasty implants using the bioelectric effect.

Introduction: The formation of bacterial biofilms on knee arthroplasty implants can have catastrophic consequences. The aim of this study was to analyze the effectiveness of the bioelectric effect in the elimination of bacterial biofilms on cultivated knee arthroplasty implants. Methods: A novel device was designed to deliver a bioelectric effect on the surface of knee arthroplasty implants. 4-femoral prosthetic implants were cultivated with a staphylococcus aureus inoculum for 15days. The components were divided into four different groups: A (not treated), B (normal saline 20-minutes), C (bioelectric effect 10-minutes), D (bioelectric effect 20-minutes). The implants were sonicated, and the detached colonies were quantified as the number of colony-forming unit (CFUs). The implants were sterilised and the process was repeated in a standardized manner four more times, to obtain a total of five samples per group. Results: The number of the CFUs after a 10-minute exposure to the bioelectric effect was of 208.2 ± 240.4, compared with 6,041.6 ± 2010.7 CFUs in group A, representing a decrease of 96.5% ± 4.3 (p = 0.004). And a diminution of 91.8% ± 7.9 compared with 2,051.0 ± 1,364.0 CFUs in group B (p = 0.109). The number of bacterial colonies after a 20-minute exposure to the bioelectric effect was 70 ± 126.7 CFUs, representing a decrease of 98.9% ± 1.9 (p = 0.000) compared with group A. And a decrease of 97.8% ± 3.0 (p = 0.019) compared with group B. Conclusions: The bioelectric effect was effective in the elimination of bacterial biofilm from knee arthroplasty implants. This method could be used in the future as part of conventional surgical procedures.

Comparative Analysis of Plaque Removal and Wear between Electric-Mechanical and Bioelectric Toothbrushes.

Effective oral care is important for maintaining a high quality of life. Therefore, plaque control can prevent the development and recurrence of periodontitis. Brushing with a toothbrush and toothpaste is a common way to remove plaque; however, excessive brushing or brushing with abrasive toothpaste can cause wear and tear on the dental crown. Hence, we aimed to quantitatively compare the plaque-removal efficiency and tooth wear of toothbrushes using the bioelectric effect (BE) with those of electric-mechanical toothbrushes. To generate the BE signal, an electronic circuit was developed and embedded in a toothbrush. Further, typodonts were coated with cultured artificial plaques and placed in a brushing simulator. A toothpaste slurry was applied, and the typodonts were eluted with tap water after brushing. The plaques of the typodonts were captured, and the images were quantified. For the tooth wear experiment, polymethyl methacrylate disk resin blocks were brushed twice a day, and the thickness of the samples was measured. Subsequently, statistical differences between the experimental toothbrushes and typical toothbrushes were analyzed. The BE toothbrush had a higher plaque-removal efficiency and could minimize tooth wear. This study suggests that the application of BE may be a new solution for oral care.

Bioelectric device for effective biofilm inflammation management of dental implants

Dental implant inflammation is primarily caused by oral biofilms, which form within 8 h, particularly at 37 °C, thereby requiring diligent cleaning. Considering the complex management of dental implants, a novel technology based on the bioelectric effect (BE) to combat inflammation has emerged. A BE-integrated toothbrush was developed and clinically tested on patients with dental implants (N = 36). Our findings revealed a significant average plaque index reduction of 67% with BE technology compared with that at baseline (P < 0.05), whereas non-BE did not yield statistical significance even after 4 weeks of use (P > 0.05). The bleeding index demonstrated a 59% average reduction in all surfaces with BE technology (P < 0.05), whereas the non-BE group exhibited no significant change. Substantial reductions in total plaque and bleeding indices suggest that using BE toothbrushes can help effectively remove oral biofilms and treat bleeding symptoms.

A Study on the Biofilm Removal Efficacy of a Bioelectric Toothbrush.

Effective oral care is a critical requirement to maintain a high quality of life. Most oral diseases are caused by plaque (oral biofilm), which is also correlated with systemic diseases. A common method to remove biofilm is brushing teeth with toothpaste. However, 3.5 billion people in the world have oral diseases, meaning that more efficient methods of removing biofilms are needed. We have developed a toothbrush that applies a bioelectric effect (BE) utilizing an electric force for biofilm removal. It demonstrated significantly higher biofilm removal efficiency than non-BE manual toothbrushes. Tests were performed in saline and toothpaste conditions using various pressures. Results showed that the BE toothbrush had a significantly higher biofilm removal efficiency in saline (0.5 N: 215.43 ± 89.92%, 2.5 N: 116.77 ± 47.02%) and in a toothpaste slurry (0.5 N: 104.96 ± 98.93%, 2.5 N: 96.23 ± 35.16%) than non-BE manual toothbrushes. Results also showed that BE toothbrushes were less dependent on toothpaste. This study suggests that the application of BE can be a new solution to plaque problems in oral care.

A photo-controlled, all-solid, and frequency-tunable ultra-wideband pulse generator.

With the continuous exploration of the bioelectric effect, nanosecond and picosecond pulsed electric fields used in cancer therapy and drug introduction have attracted great attention. In this paper, an ultrashort pulsed electric field generator is proposed, which connects two photoconductive semiconductor switches in parallel to generate unipolar and bipolar pulses. We described the experimental scheme of the generator and the simulation of the radio frequency combiner. A 532nm laser with pulse widths of 1ns and 500ps is used to trigger the photoconductive semiconductor switches. The experimental results show that the scheme can achieve adjustments of 357 and 720MHz for the center frequency and the 3 dB bandwidth, respectively. The results confirm that this proposed scheme can be used for unipolar/bipolar frequency-adjustable ultra-wideband pulse generation.

An Experimental Study of Non-Electrolysis Anti-Microfouling Technology Based on Bioelectric Effect

Biofouling initiated by biofilm (slime) formation is a key challenge for practical ocean engineering and construction. This study evaluated a new anti-biofilm technology using bioelectricity. The anti-microfouling electrical technology is based on the principles of the bioelectric effect, known as the application of an electrostatic force for biofilm removal. Previously, the electricity was optimized below 0.82V to avoid electrolysis, which can prevent the production of biocides. A test boat comprised of microelectronics for electrical signal generation with electrodes for an anti-biofouling effect was developed. The tests were conducted in the West Sea of Korea (Wangsan Marina, Incheon) for three weeks. The surface biofouling was quantified. A significant reduction of fouling was observed under the bioelectric effect conditions, with approximately 30% enhanced prevention of fouling progress (P&lt;0.05). This technology can be an alternative eco-friendly technique for anti-microfouling that can be applied for canals, vessels, and coastal infrastructure because it does not induce electrolysis.

Simulation and Experimental Study on the Responses of Subcellular Structures in Tumor Cells Induced by 5 ns Pulsed Electric Fields

In order to explore the bioelectric effect of 5 ns pulsed electric fields on tumor cells, a spherical single-cell multiphysics model was first established based on the finite element simulation platform. In consideration of the dielectric relaxation of the biological plasma membrane under the high-frequency electric fields, the electroporation and Maxwell stress tensors on the cell membrane and nuclear envelope were analyzed; secondly, taking MDA-MB-231 cells as the research object, combined with fluorescent probe technology, the state change and fluorescence dissipation of its subcellular structure exposed to pulse fields were studied. The results showed that 5 ns pulsed electric fields directly acted inside the cell, causing an electroporation effect and tensile stress on the nuclear envelope, destroying the integrity and order of the cytoskeleton, and damaging the functions of subcellular structures including endoplasmic reticulum, mitochondria, etc. This study provides theoretical and experimental evidence for the research and application of a high-voltage short pulse in the field of biomedical engineering.

A Non-Electrolysis Bioelectric Effect for Gingivitis and Hygiene Contamination Biofilm Removal

A combinatorial biofilm treatment involving a low dose of antibiotics along with a small amount of electricity is known as the bioelectric effect (BE). When an external electric field or current is applied, biofilms can be affected by the resulting electrostatic force. Our group is researching the maximization of electrostatic force through the integration of both alternating and direct currents, with a voltage below the electrolysis threshold (0.82 V). To validate the efficacy of this technology, in the present work we investigated two major biofilm applications: (1) dentistry for oral biofilm infection and (2) hygiene for aerobic biofilm contamination. For each application, testing devices were developed in the form of a toothbrush and an evaporator cleaner, respectively. The dental clinical results demonstrated a 75% reduction in gingivitis compared to the non-BE applied group (n = 40, ANOVA, paired t-test, p &lt; 0.05). Meanwhile, the hygiene testing result demonstrated an 81.8% increase in biofilm removal compared to the initial untreated sample (n = 6, ANOVA, paired t-test, p &lt; 0.05). In conclusion, this new BE technology showed efficacy in both dental- and hygiene-associated biofilms without causing electrolysis. Further investigation and development of the BE system should continue in both the medical and hygiene fields.

Developing an Electrical System toward the Prevention of Heat Ventilation Air Conditioning Contamination

Biofilms comprise diverse bacteria and fungi with slime material, especially in humid conditions. The surface of the heat ventilation and air conditioning (HVAC) system, especially at the evaporator, is in high moisture conditions due to the compression and relaxation of refrigerant. Biofilms comprise electrically polarized molecules, which include polysaccharide, DNA, liposome, and diverse amino acids. Thus, when an electric field is applied to biofilms, the field creates an electrostatic force that enhances the destruction of biofilms and results in detachment from the surface. The principles are known as the bioelectric effect. A new bioelectric (BE) technology has been developed that is focused on the optimization of electrical energy for biofilm removal, and its efficacy has been previously demonstrated. In this work, a prototype of the BE device for HVAC application has been fabricated and tested. Escherichia coli (E. coli) biofilms were utilized as a standard model of biofilm on the HVAC evaporator. The results demonstrate that when the BE technology was applied for one hour, approximately 81.8% of total biofilm was removed. Furthermore, live cells in biofilms were reduced by approximately 71.4% under the BE device, compared to non-treated biofilms.

Bioelectric effect utilized a healthcare device for effective management of dental biofilms and gingivitis

Oral healthcare is a critical condition for high quality of life. According to statistics, gingivitis is one of major diseases in the world (The Global Burden of Disease Study 2019, 3.5 billion population worldwide). Oral diseases, that initiated by a group of multispecies bacteria with diverse extracellular slime that called biofilms, are modifiable risk factors with the cancer, cardiovascular disease, chronic respiratory disease. Hence, effective management of biofilms is important.Since biofilms are comprised of extracellular matrix that inhibit drug diffusion, the traditional antibiotic treatment requires 500 -5000 times more concentration than planktonic bacterial infection. The biofilm cleaning in dentistry includes time-consuming and painful physical brushing.In this work, we have developed a new electric toothbrush in emphasis on demonstration of convenient and painless gingivitis improvement device. For the first time, the toothbrush has been integrated with an electromagnetic technology for effective biofilm removal, known for bioelectric effect (BE). Biofilms consists of electrically charged molecules that are affected by external electric fields. Thus, when an electric field is applied, it creates interruption of metabolism, reduction of enzyme activity, weakening surface attachment, and nonuniform electrolyte distribution, resulting in separation from the surface. Thus, integration of the BE on a toothbrush enables enhanced biofilm reduction.The toothbrush tested with mild gingival patient (N=40) at the department of dentistry, Ulsan University Hospital, South Korea. The patient used the BE toothbrush and non-BE toothbrush consecutively for 2 weeks in each, following their gingival index (GI) and plaque index (PI) measurement for the quantitative analysis. The results demonstrate 1.75 times more decrease of GI in BE toothbrush than the data of the non-BE device (p < 0.05).In conclusion, a new electromagnetic wave toothbrush has been successfully demonstrated especially in reduction of inflammation. Based on this work, we are going to further develop a medical device that dedicated on the treatment of inflammation.

Bioelectric Effect of a Microcurrent Toothbrush on Plaque Removal.

This study evaluated the effectiveness of a microcurrent toothbrush (approved by the US Food and Drug Administration [FDA]), which employs a superimposed alternating and direct electric current, named as a Proxywave® technology, similar to the intensity of the biocurrent, in plaque removal and reducing gingivitis by biofilm removal through the bioelectric effect. This study enrolled 40 volunteers with gingivitis. Dental observations were made every two weeks, before and after the use of each toothbrush. We randomly assigned participants into two groups: one group used the Proxywave® toothbrush (PB) for two weeks followed by the control toothbrush (CB) for two weeks, while the other group used the CB for two weeks followed by the PB. The participants had a two-week washout period. If the toothbrush used earlier has had an effect on the bacterial flora in the oral cavity, this is to remove this effect and return it to its previous state. During each dental visit, we recorded plaque index (PI) and gingival index (GI) scores. The PI and GI scores were significantly lower in both the PB and the CB (p < 0.05). Considering the PI, there was no significant difference between the toothbrushes on all the surfaces. Considering the GI, the PB showed a significant decrease in the interproximal surface, compared to the CB (p < 0.05). The PB showed a significant decrease in the interproximal GI and had a beneficial effect in the interproximal area where the bristles could not reach. No adverse events were observed in the participants during the clinical trial. The microcurrent toothbrush is a device that can be safely used for plaque removal.

Electroactive 3D Printed Scaffolds Based on Percolated Composites of Polycaprolactone With Thermally Reduced Graphene Oxide for Antibacterial and Tissue Engineering Applications.

Applying electrical stimulation (ES) could affect different cellular mechanisms, thereby producing a bactericidal effect and an increase in human cell viability. Despite its relevance, this bioelectric effect has been barely reported in percolated conductive biopolymers. In this context, electroactive polycaprolactone (PCL) scaffolds with conductive Thermally Reduced Graphene Oxide (TrGO) nanoparticles were obtained by a 3D printing method. Under direct current (DC) along the percolated scaffolds, a strong antibacterial effect was observed, which completely eradicated S. aureus on the surface of scaffolds. Notably, the same ES regime also produced a four-fold increase in the viability of human mesenchymal stem cells attached to the 3D conductive PCL/TrGO scaffold compared with the pure PCL scaffold. These results have widened the design of novel electroactive composite polymers that could both eliminate the bacteria adhered to the scaffold and increase human cell viability, which have great potential in tissue engineering applications.

Flexible Platform for In Situ Impedimetric Detection and Bioelectric Effect Treatment of Escherichia Coli Biofilms.

This paper reports a platform for real-time monitoring and treatment of biofilm formation on three-dimensional biomedical device surfaces. We utilize a flexible platform consisting of gold interdigitated electrodes patterned on a polyimide substrate. The device was integrated onto the interior of a urinary catheter and characterization was performed in a custom-developed flow system. Biofilm growth was monitored via impedance change at 100Hz ac with a 50mV signal amplitude. A 30% impedance decrease over 24h corresponded to Escherichia coli biofilm formation. The platform also enabled removal of the biofilm through the bioelectric effect; a low concentration of antibiotic combined with the applied ac voltage signal led to a synergistic reduction in biofilm resulting in a 12% increase in impedance. Biomass characterization via crystal violet staining confirmed that the impedance detection results correlate with changes in the amount of biofilm biomass on the sensor. We also demonstrated integration with a chip-based impedance converter to enable miniaturization and allow in situ wireless implementation. A 5% impedance decrease measured with the impedance converter corresponded to biofilm growth, replicating the trend measured with the potentiostat. This platform represents a promising solution for biofilm infection management in diverse vulnerable environments. Biofilms are the dominant mode of growth for microorganisms, where bacterial cells colonize hydrated surfaces and lead to recurring infections. Due to the inaccessible nature of the environments where biofilms grow and their increased tolerance of antimicrobials, identification, and removal on medical devices poses a challenge.

Application of bioelectric effect to reduce the antibiotic resistance of subgingival plaque biofilm: An in vitro study.

Context:Biofilms are known for their antimicrobial resistance, and so is the subgingival plaque biofilm, the primary etiologic factor for periodontal infections.Aims:The objective of this study is to investigate if the subgingival plaque biofilm resistance can be reduced using doxycycline in the presence of low-intensity electric field (bioelectric effect).Settings and Design:The study was an in vitro microbiological study.Materials and Methods:Subgingival plaque samples from chronic periodontitis patients were collected to grow subgingival plaque biofilms on hydroxyapatite disks. Hydroxyapatite disks with the plaque biofilms from each patient were divided into four groups: (i) No intervention – control, (ii) current alone – CU; (iii) doxycycline – AB, and (iv) combined treatment – CU + AB. After respective treatments, the disks were anaerobically incubated for 48 h, the biofilm was dispersed and subcultured and colony-forming unit/mL was estimated in all the four groups.Statistical Analysis:Statistical analysis was done using Mann–Whitney and Kruskal–Wallis tests for intergroup comparisons. T-test was done to assess the difference in current flow between the groups CU and CU + AB.Results:All the three treatment modalities showed antibacterial effect. Application of current alone resulted in reduced bacterial growth than control group. Doxycycline alone resulted in reduction in bacterial counts better than control and current alone groups. The combination treatment showed greatest inhibition of bacterial colonies.Conclusion:The ability of doxycycline antibiotic in inhibiting plaque biofilm was significantly enhanced by application of a weak electric field (5 volts for 2 min).

Effect of electric currents on antibacterial effect of chlorhexidine against Entrococcus faecalis biofilm: An in vitro study.

BackgroundThis in vitro study was mainly aimed to evaluate the effect of high-frequency alternating currents (AC) applied by an electronic apex locator (EAL) on the antibacterial properties of chlorhexidine (CHX) on E. faecalis biofilm.Material and MethodsThe root canals of 120 extracted human single-rooted teeth were prepared using Gates-Glidden drills and hand K-files. After contaminating the root canals with E. faecalis, they were incubated for 60 days. Then, the teeth were randomly divided into six experimental groups (n=20). Group 1, 2% CHX; group 2, normal saline (NS) with direct current (DC); group 3, normal saline (NS) with high-frequency alternating current (AC); group 4, 2% CHX with DC; group 5, 2% CHX with AC; group 6, control (normal saline). The samples were collected from the root canal walls of 16 teeth in each group and 1:10 serial dilutions were prepared and added to Muller-Hinton agar (MHA) plates and incubated at 37°C for 48 h. The longitudinal sections of the other 4 teeth used to observe under a scanning electron microscope (SEM). A classic colony counting technique was used for counting the vital E. faecalis bacteria in MHA. Two-way ANOVA was used for statistical analysis of the data. The level of significance was set at P<0.05.ResultsThe electric current significantly changed the colony-forming units (CFU) values (P<0.001). According to pair-wise comparisons, the highest CFU difference was observed between the AC group and the group without electric current (P<0.001); furthermore, the difference between the DC group and the group without electric current was not significant (P=0.823).ConclusionsThe highest bioelectric effect occurred with the use of high-frequency alternating electric current in the form of an apex locator with CHX as a canal irrigant. Key words:Biofilm, Chlorhexidine, Direct current, Electric current, Enterococcus faecalis.

An Integrated Microsystem for Real-Time Detection and Threshold-Activated Treatment of Bacterial Biofilms.

Bacterial biofilms are the primary cause of infections in medical implants and catheters. Delayed detection of biofilm infections contributes to the widespread use of high doses of antibiotics, leading to the emergence of antibiotic-resistant bacterial strains. Accordingly, there is an urgent need for systems that can rapidly detect and treat biofilm infections in situ. As a step toward this goal, in this work we have developed for the first time a threshold-activated feedback-based impedance sensor-treatment system for combined real-time detection and treatment of biofilms. Specifically, we demonstrate the use of impedimetric sensing to accurately monitor the growth of Escherichia coli biofilms in microfluidic flow cells by measuring the fractional relative change (FRC) in absolute impedance. Furthermore, we demonstrate the use of growth measurements as a threshold-activated trigger mechanism to initiate successful treatment of biofilms using bioelectric effect (BE), applied through the same sensing electrode array. This was made possible through a custom program that (a) monitored the growth and removal of biofilms within the microfluidic channels in real-time and (b) enabled the threshold-based activation of BE treatment. Such BE treatment resulted in a ∼74.8 % reduction in average biofilm surface coverage as compared to the untreated negative control. We believe that this smart microsystem for integrated biofilm sensing and treatment will enable future development of autonomous biosensors optimized for accurate real-time detection of the onset of biofilms and their in situ treatment, directly on the surfaces of medical implants.

In Vitro Assessment of Electric Currents Increasing the Effectiveness of Vancomycin Against Staphylococcus epidermidis Biofilms.

Biofilms are communities of bacteria that can cause infections which are resistant to the immune system and antimicrobial treatments, posing a significant threat for patients with implantable and indwelling medical devices. The purpose of our research was to determine if utilizing specific parameters for electric currents in conjunction with antibiotics could effectively treat a highly resistant biofilm. Our study evaluated the impact of 16 μg/mL of vancomycin with or without 22 or 333 μA of direct electric current (DC) generated by stainless steel electrodes against 24-, 48-, and 72-h-old Staphylococcus epidermidis biofilms formed on titanium coupons. An increase in effectiveness of vancomycin was observed with the combination of 333 μA of electric current against 48-h-old biofilms (P value = 0.01) as well as in combination with 22 μA of electric current against 72-h-old biofilms (P value = 0.04); 333 μA of electric current showed the most significant impact on the effectiveness of vancomycin against S. epidermidis biofilms demonstrating a bioelectric effect previously not observed against this strain of bacteria.

An optical microfluidic platform for spatiotemporal biofilm treatment monitoring

Bacterial biofilms constitute in excess of 65% of clinical microbial infections, with the antibiotic treatment of biofilm infections posing a unique challenge due to their high antibiotic tolerance. Recent studies performed in our group have demonstrated that a bioelectric effect featuring low-intensity electric signals combined with antibiotics can significantly improve the efficacy of biofilm treatment. In this work, we demonstrate the bioelectric effect using sub-micron thick planar electrodes in a microfluidic device. This is critical in efforts to develop microsystems for clinical biofilm infection management, including both in vivo and in vitro applications. Adaptation of the method to the microscale, for example, can enable the development of localized biofilm infection treatment using microfabricated medical devices, while augmenting existing capabilities to perform biofilm management beyond the clinical realm. Furthermore, due to scale-down of the system, the voltage requirement for inducing the electric field is reduced further below the media electrolysis threshold. Enhanced biofilm treatment using the bioelectric effect in the developed microfluidic device elicited a 56% greater reduction in viable cell density and 26% further decrease in biomass growth compared to traditional antibiotic therapy. This biofilm treatment efficacy, demonstrated in a micro-scale device and utilizing biocompatible voltage ranges, encourages the use of this method for future clinical biofilm treatment applications.

A surface acoustic wave biofilm sensor integrated with a treatment method based on the bioelectric effect

Bacterial biofilms have an extensive impact on quality of life, ranging from severe infections in the clinical field to water facility contamination in environmental science. Biofilms are comprised of diverse bacteria that produce an extracellular matrix which prevents drug diffusion through them. Hence traditional antibiotic therapies require 500–5000 times the concentration used to eliminate non-biofilm-associated infections. Early biofilm detection is critical for effective eradication. Moreover, developing an alternative biofilm treatment method that utilizes low doses of antibiotics is desired. In this paper, a real-time microsystem is shown to detect growth of biofilms as well as their removal through integrated treatment. Detection of biofilms is achieved using a surface acoustic wave (SAW) sensor that monitors the total biomass by measuring the resonant frequency of the system. Biofilm treatment is based on the bioelectric effect (BE), a combination of low-dose antibiotics with application of both alternating and direct current signals. The detection limit of the SAW system is approximately 166pg, corresponding to a bacterial population on the order of hundreds of bacteria. The system is used to observe an 80% reduction of total biomass when treated by the BE as compared to traditional antibiotics. Through system integration of the BE with the SAW sensor, simultaneous biofilm detection and treatment is achieved. The system consumes 194μW of power, with the sensor and treatment consuming 100μW and 94μW, respectively. The integrated sensing and treatment capabilities of this system advance the development of an innovative biofilm control method.

Effect of electrical energy on the efficacy of biofilm treatment using the bioelectric effect.

Background/Objectives:The use of electric fields in combination with small doses of antibiotics for enhanced treatment of biofilms is termed the ‘bioelectric effect’ (BE). Different mechanisms of action for the AC and DC fields have been reported in the literature over the last two decades. In this work, we conduct the first study on the correlation between the electrical energy and the treatment efficacy of the bioelectric effect on Escherichia coli K-12 W3110 biofilms.Methods:A thorough study was performed through the application of alternating (AC), direct (DC) and superimposed (SP) potentials of different amplitudes on mature E. coli biofilms. The electric fields were applied in combination with the antibiotic gentamicin (10 μg/ml) over a course of 24 h, after the biofilms had matured for 24 h. The biofilms were analysed using the crystal violet assay, the colony-forming unit method and fluorescence microscopy.Results:Results show that there is no statistical difference in treatment efficacy between the DC-, AC- and SP-based BE treatment of equivalent energies (analysis of variance (ANOVA) P>0.05) for voltages <1 V. We also demonstrate that the efficacy of the BE treatment as measured by the crystal violet staining method and colony-forming unit assay is proportional to the electrical energy applied (ANOVA P<0.05). We further verify that the treatment efficacy varies linearly with the energy of the BE treatment (r2 =0.984). Our results thus suggest that the energy of the electrical signal is the primary factor in determining the efficacy of the BE treatment, at potentials less than the media electrolysis voltage.Conclusions:Our results demonstrate that the energy of the electrical signal, and not the type of electrical signal (AC or DC or SP), is the key to determine the efficacy of the BE treatment. We anticipate that this observation will pave the way for further understanding of the mechanism of action of the BE treatment method and may open new doors to the use of electric fields in the treatment of bacterial biofilms.