Bactericidal effect of nanostructures <i>via</i> lytic transglycosylases of <i>Escherichia coli</i>

Mimura, Soma; Shimizu, Tomohiro; Shingubara, Shoso; Iwaki, Hiroaki; Ito, Tetsuo

doi:10.1039/d1ra07623j

Cited by 12 publications

(8 citation statements)

References 43 publications

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“…To confirm the cell membrane damage, we analyzed the ratio of green-stained cells versus whole cells (green- and red-stained cells) on the WE using PI and SYTO 9 reagents. This experiment was conducted using the fluorescence microscope . The green-stained cell showed that its membrane was not damaged.…”

Section: Resultsmentioning

confidence: 99%

“…Ishak et al suggested that deformation of the cell envelope was attributed to the surface-sensing system of bacterium due to surface protein such as phase shock protein A (PspA) . We previously reported that autolysis may also contribute to an organism’s death on nanostructured surfaces; in this mechanism, the physical interaction with a nanostructure serves as the primary trigger and autolysis activation serves as the secondary trigger, causing the cell to lyse itself . In this study, we sought to determine the veracity of this theory by experimentally confirming whether the simplest and weakest type of lipid bilayer could be disrupted by physical factors alone.…”

Section: Discussionmentioning

confidence: 95%

“…Many scientists have investigated the bactericidal mechanisms of nanostructures from the theoretical and experimental viewpoints. The relevant studies have revealed that nanostructures have four possible mechanisms: (i) direct penetration into the bacterial cell envelope, (ii) cell envelope stretching and rupture between nanostructures, , (iii) envelope deformation due to interactions between bacterial cells and nanostructures, and (iv) biochemical reactions due to surface protein , or autolytic enzyme activation after physical deformation . In all of these cases, the bacteria–nanopattern interaction is certainly the first trigger.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescence Intensity of Liposomes and E. coli Attached to Nanopillar Arrays: Implications for Bacterial Death on Nanostructures

Daimon

Yanagisawa

Ishibashi

et al. 2023

ACS Appl. Nano Mater.

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Bacterial death on nanostructures has sparked great interest because the principle behind this phenomenon completely differs from that of conventional chemical antimicrobial agents. Physical interactions between the bacterial envelope and nanostructures generate forces that deform the cell membrane. Eventually, the cell envelope breaks, causing the cytoplasm to leak out. However, the events occurring between these stages are unclear. In this work, we used liposomes as pseudocells because they are composed of a lipid bilayer, which is similar to a cytoplasmic membrane. We monitored time-dependent changes in the fluorescence intensities of rhodamine 6G (R6G)-containing liposomes and mCherry-expressing Escherichia coli cells after adhering to Au nanostructure arrays (pitch; height; diameter = 500, 300, 283 nm) and compared the variations in the fluorescence intensities of the pseudo and E. coli cells over time. Electrochemical impedance spectroscopy was then performed to monitor the cell and liposome attachment. Ratio of change in R ct before and after the liposome injection was about 1.7, which means that R ct increased drastically due to the attachment of liposome. Similarly, ratio of change in R ct before and after the E. coli injection was about 1.15 due to the attachment of cell. The fluorescence intensity of mCherry quickly decreased after cell adhesion within 25 min, whereas that of R6G did not even at 60 min. This finding means that the liposomes were not broken down despite their mechanical strength being equal to or weaker than that of E. coli. The time constant (T) of the decrease in fluorescence intensity of E. coli was calculated by fitting the data to an exponential function and found to be −0.13 min −1 , which is in good agreement with our previous results. But we could not calculate the T of liposomes. The results indicate that the death of bacteria on nanostructures likely involves an autolytic mechanism.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

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Fluorescence Intensity of Liposomes and E. coli Attached to Nanopillar Arrays: Implications for Bacterial Death on Nanostructures

Daimon

Yanagisawa

Ishibashi

et al. 2023

ACS Appl. Nano Mater.

Self Cite

View full text Add to dashboard Cite

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“…On the other hand, sharp, spiky-like nanofeatures can pierce, or those with sharp edges (e.g., graphene flakes, 2 nm) can cut the cell membrane. 7,8 Thus, nanostructured surfaces capable of deforming cell membranes have become a more promising surface design for preventing the spread of pathogenic bacteria, which can improve bactericidal efficiency. An overly complex process of cell−nanostructure interaction strongly depends on the surface properties of the bacterium.…”

Section: Introductionmentioning

confidence: 99%

Bactericidal Performance of Nanostructures Resin Surfaces: Nanopillars versus Nanocones

Zhao,

Matsushita,

Ogawa

et al. 2024

ACS Appl. Nano Mater.

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“…The bactericidal mechanism is complex and several explanations have been advanced. While bacterial death when in contact with nanostructures is often attributed to membrane rupture, some studies have demonstrated that physical stress from membrane deformation without rupture leads to increased reactive oxygen species production and autolytic activity. , A recent review on the topic concluded that pillar spacing needs to be smaller than the size of the bacteria, therefore sufficiently small to prohibit bacterial attachment in between pillars. Furthermore, if pillars are short and sparse, the bacterial membrane needs to stretch less until it covers the substrate between pillars.…”

Section: Introductionmentioning

confidence: 99%

Effect of Flexibility and Size of Nanofabricated Topographies on the Mechanobactericidal Efficacy of Polymeric Surfaces

Lohmann

Tripathy

Milionis

et al. 2022

ACS Appl. Bio Mater.

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Driven by the growing threat of antimicrobial resistance, the design of intrinsically bactericidal surfaces has been gaining significant attention. Proposed surface topography designs are often inspired by naturally occurring nanopatterns on insect wings that mechanically damage bacteria via membrane deformation. The stability of and the absence of chemicals in such surfaces support their facile and sustainable employment in avoiding surface-born pathogen transmission. Recently, the deflection of controllably nanofabricated pillar arrays has been shown to strongly affect bactericidal activity, with the limits of mechanical effectiveness of such structures remaining largely unexplored. Here, we examine the limits of softer, commonly used polymeric materials and investigate the interplay between pillar nanostructure sizing and flexibility for effective antibacterial functionality. A facile, scalable, UV nanoimprint lithography method was used to fabricate nanopillar array topographies of variable sizes and flexibilities. It was found that bacterial death on nanopillars in the range of diameters ≤100 nm and Young’s moduli ≥1.3 GPa is increased by 3.5- to 5.6-fold, while thicker or softer pillars did not reduce bacterial viability. To further support our findings, we performed a finite element analysis of pillar deformation. It revealed that differences in the amount of stress exerted on bacterial membranes, generated from the stored elastic energy in flexible pillars, contribute to the observed bactericidal performance.

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Bactericidal effect of nanostructures via lytic transglycosylases of Escherichia coli

Abstract: The time profiles of active cell ratios depended on the growth phase and the absence of some lytic transglycosylases of E. coli. Significant cell damage was not found on the autolysis inhibition condition.

Cited by 12 publications

References 43 publications

Fluorescence Intensity of Liposomes and E. coli Attached to Nanopillar Arrays: Implications for Bacterial Death on Nanostructures

Fluorescence Intensity of Liposomes and E. coli Attached to Nanopillar Arrays: Implications for Bacterial Death on Nanostructures

Bactericidal Performance of Nanostructures Resin Surfaces: Nanopillars versus Nanocones

Effect of Flexibility and Size of Nanofabricated Topographies on the Mechanobactericidal Efficacy of Polymeric Surfaces

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