A. James Clark School of Engineering
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Item Effect of electrical energy on the efficacy of biofilm treatment using the bioelectric effect(Nature Publishing Group, 2015-09-23) Kim, Young Wook; Subramanian, Sowmya; Gerasopoulos, Konstantinos; Ben-Yoav, Hadar; Wu, Hsuan-Chen; Quan, David; Carter, Karen; Meyer, Mariana T.; Bentley, William E.; Ghodssi, RezaBACKGROUND/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.Item AN INTEGRATED MICROSYSTEM FOR BACTERIAL BIOFILM DETECTION AND TREATMENT(2014) Kim, Young Wook; Ghodssi, Reza; Electrical Engineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Bacterial biofilms cause severe infections in clinical fields and contamination problems in environmental facilities. Due to the unique complex structure of biofilms that comprise diverse polysaccharides and bacteria, traditional antibiotic therapies require a thousand times higher concentration compared to non-biofilm associated infections. The early detection of biofilms, before their structures are fully established in a given host/environment, is critical in order to eradicate them effectively. Also, the development of a new innovative biofilm treatment method that can be utilized with a low dose of antibiotic would be extremely important to the medical community. In this dissertation, a biofilm sensor and a new biofilm treatment method were independently developed to detect and treat biofilm communities, respectively. Furthermore, an integrated microsystem was demonstrated as a single platform of the sensor with the treatment method. The sensor was based on the surface acoustic wave (SAW) detection mechanism, which isn extremely sensitive for biofilm monitoring (hundreds of bacterial population detection limit) and consumes very low power (~100 µW). A piezoelectric ZnO layer fabricated by a pulsed laser deposition process was a key material to induce homogeneous acoustic waves. Reliable operation of the sensor was achieved using an Al2O3 film as a passivation layer over the sensor to protect ZnO degradation from the growth media. The sensor successfully demonstrated real-time monitoring of biofilm growth. The new biofilm treatment was developed based on the principles of the bioelectric effect that introduces an electric field along with antibiotics to biofilms, demonstrating significant biofilm inhibition compared to antibiotic treatment alone. Specifically, the new bioelectric effect was implemented with a superpositioned (SP) electric field of both alternating and direct current (AC and DC) and the antibiotic gentamicin (10 µg/mL). With the SP field treatment, significant biofilm reduction was demonstrated in total biomass (~ 71 %) as well as viable bacterial density (~ 400 times respected to the only antibiotic therapy) of the treated biofilms. This method was transferred to a microfluidic system using microfabricated planar electrodes. The microsystem-level implementation of the bioelectric effect also showed enhanced biofilm reduction (~ 140 % total biomass reduction improvement). The integrated system was based on the SAW sensor with the addition of coplanar thin electrodes to apply electric signals for the biofilm treatment. The chip was tested with two bacterial biofilms (Escherichia coli and Pseudomonas aeruginosa) that are clinically relevant strains. In both biofilm experiments, the integrated system demonstrated successful real-time biofilm monitoring and effective biofilm inhibition. This systematic integration of a continuous monitoring method with a novel effective treatment technique is expected to advance the state of the art in the field of managing clinical and environmental biofilms.Item AN ATOMIC LAYER DEPOSITION PASSIVATED SURFACE ACOUSTIC WAVE SENSOR FOR BACTERIAL BIOFILM GROWTH MONITORING(2012) Kim, Young Wook; Ghodssi, Reza; Electrical Engineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)This thesis reports for the first time the design, fabrication, and testing of a reusable Surface Acoustic Wave (SAW) sensor for biofilm growth monitoring. Bacterial biofilms cause severe infections, and are often difficult to remove without an invasive surgery. Thus, their detection at an early stage is critical for effective treatments. A highly sensitive SAW sensor for biofilm growth monitoring was fabricated by depositing a high quality zinc oxide (ZnO) piezoelectric thin film by pulsed laser deposition (PLD). The sensor was successfully passivated by aluminum oxide (Al2O3) using Atomic Layer Deposition (ALD) to prevent ZnO damage from long term media contact. The sensor was reusable over multiple biofilm formation experiments using the ALD Al2O3 passivation and an oxygen plasma biofilm cleaning method. The SAW sensor was studied with Escherichia coli biofilm growth in Lysogeny Broth (LB) and in 10% Fetal Bovine Serum (FBS) as a simulated an in vivo environment. A multiple MHz level resonant frequency shift measured at the output of the SAW sensor in both LB and 10% FBS corresponded to the natural biofilm growth progression. These repeatable E. coli biofilm growth monitoring results validate the novel application of a SAW sensor for future implantable biofilm sensing applications.