How Much Force is Needed to Kill a Single Bacterium?

The interaction between bacteria and nanomaterials, particularly from a physical or mechanical perspective, has emerged as a topic of significant interest in both science and medicine. Mechanobactericidal nanomaterials, which exert antimicrobial effects through purely physical mechanisms, hold promise as alternative strategies to combat bacterial resistance to traditional antibiotics. High-aspect-ratio nanoparticles and surface topographies are being engineered to enhance their mechanobactericidal properties.

Anti-Biofilm Perspectives of Propolis against Infections.

has emerged as the main causative agent of medical device-related infections. Their major pathogenicity factor lies in its ability to adhere to surfaces and proliferate into biofilms, which increase their resistance to antibiotics. The main objective of this study was to evaluate the use and the mechanism of action of an ethanolic extract of Spanish propolis (EESP) as a potential alternative for preventing biofilm-related infections caused by .

Antimicrobial activity of a novel Spanish propolis against planktonic and sessile oral Streptococcus spp.

Increased bacterial resistance to traditional antimicrobial agents has prompted the use of natural products with antimicrobial properties such as propolis, extensively employed since ancient times. However, the chemical composition of propolis extracts is extremely complex and has been shown to vary depending on the region and season of collection, due to variations in the flora from which the pharmacological substances are obtained, being therefore essential for their antimicrobial activity to be checked before use. For this purpose, we evaluate the in vitro antimicrobial and anti-biofilm activity of a new and promising Spanish ethanolic extract of propolis (SEEP) on Streptococcus mutans and Streptococcus sanguinis, responsible, as dominant 'pioneer' species, for dental plaque.

Decomposition of Growth Curves into Growth Rate and Acceleration: a Novel Procedure To Monitor Bacterial Growth and the Time-Dependent Effect of Antimicrobials.

In this paper, a simple numerical procedure is presented to monitor the growth of Streptococcus sanguinis over time in the absence and presence of propolis, a natural antimicrobial. In particular, it is shown that the real-time decomposition of growth curves obtained through optical density measurements into growth rate and acceleration can be a powerful tool to precisely assess a large range of key parameters (i.e.

A physico-chemical study of the interaction of ethanolic extracts of propolis with bacterial cells.

In the present study, an effort has been made to understand the interaction mode of propolis, a natural substance produced by honey bees, with gram-positive and gram-negative bacterial cells by measuring alterations in cell surface physico-chemical properties following the incubation of the cells with different sub-inhibitory concentrations of this antimicrobial agent. Electrophoretic mobility and surface hydrophobicity measurements revealed for the first time that propolis induced substantial changes in the volumetric charge density, electrophoretic softness and degree of hydrophobicity characterizing the outermost surface layer of cells. These changes, which appear to be dose-dependent, seem to be consistent with the increasing accumulation and penetration of the propolis antimicrobial components through the cells extracellular layer.

Mechanically Induced Bacterial Death Imaged in Real Time: A Simultaneous Nanoindentation and Fluorescence Microscopy Study.

Mechano-bactericidal nanomaterials rely on their mechanical or physical interactions with bacteria and are promising antimicrobial strategies that overcome bacterial resistance. However, the real effect of mechanical versus chemical action on their activity is under debate. In this paper, we quantify the forces necessary to produce critical damage to the bacterial cell wall by performing simultaneous nanoindentation and fluorescence imaging of single bacterial cells.

Bacterial response to spatially organized microtopographic surface patterns with nanometer scale roughness.

In this study, the influence of nanometer scale roughness on bacterial adhesion and subsequent biofilm formation has been evaluated using spatially organized microtopographic surface patterns for four major opportunistic pathogens of the genus Staphylococcus (S. epidermidis and S. aureus) responsible for associated-biofilm infections on biomedical devices.

On the interactions of human bone cells with Ti6Al4V thermally oxidized by means of laser shock processing.

We investigated a Ti6Al4V alloy modified by means of laser peening in the absence of sacrificial coatings. As a consequence of the temperature rise during laser focusing, melting and ablation generated an undulated surface that exhibits an important increase in the content of titanium oxides and OH- ions. Human mesenchymal stem cells and osteoblasts cultured on the oxidized alloy develop noticeable filopodia and lamellipodia.

Studying the influence of surface topography on bacterial adhesion using spatially organized microtopographic surface patterns.

The influence of surface topography on bacterial adhesion has been investigated using a range of spatially organized microtopographic surface patterns generated on polydimethylsiloxane (PDMS) and three unrelated bacterial strains. The results presented indicate that bacterial cells actively choose their position to settle, differentiating upper and lower areas in all the surface patterns evaluated. Such selective adhesion depends on the cells' size and shape relative to the dimensions of the surface topographical features and surface hydrophobicity/hydrophilicity.

Electrochemical analysis of the UV treated bactericidal Ti6Al4V surfaces.

This research investigates in detail the bactericidal effect exhibited by the surface of the biomaterial Ti6Al4V after being subjected to UV-C light. It has been recently hypothesized that small surface currents, occurring as a consequence of the electron-hole pair recombination taking place after the excitation process, are behind the bactericidal properties displayed by this UV-treated material. To corroborate this hypothesis we have used different electrochemical techniques, such as electrochemical impedance spectroscopy (EIS), potentiodynamic polarization plots and Mott-Schottky plots.

Surface-dependent mechanical stability of adsorbed human plasma fibronectin on Ti6Al4V: domain unfolding and stepwise unraveling of single compact molecules.

In this study, the structure and mechanical stability of human plasma fibronectin (HFN), a major protein component of blood plasma, have been evaluated in detail upon adsorption on the nonirradiated and irradiated Ti6Al4V material through the use of atomic force microscopy. The results indicated that the material surface changes occurring after the irradiation process reduce the disulfide bonds that typically preclude the mechanical denaturation of individual HFN domains and interfere significantly with the intraionic interactions stabilizing the compact conformation of the adsorbed HFN molecules. In particular, upon adsorption on this material, the molecules adopt a more flexible conformation and become mechanically more compliant.

Adsorption behavior of human plasma fibronectin on hydrophobic and hydrophilic Ti6Al4V substrata and its influence on bacterial adhesion and detachment.

Biomaterial implant-associated infections, a common cause of medical devices' failure, are initiated by bacterial adhesion to an adsorbed protein layer on the implant material surface. In this study, the influence of protein surface orientation on bacterial adhesion has been examined using three clinically relevant bacterial strains known to express specific binding sites for human plasma fibronectin (HFN). HFN was allowed to adsorb on hydrophobic Ti6Al4V and physically modified hydrophilic Ti6Al4V substrata.

The zeta potential of extended dielectrics and conductors in terms of streaming potential and streaming current measurements.

The electrical characterization of surfaces in terms of the zeta potential (ζ), i.e., the electric potential contributing to the interaction potential energy, is of major importance in a wide variety of industrial, environmental and biomedical applications in which the integration of any material with the surrounding media is initially mediated by the physico-chemical properties of its outer surface layer.

Insights into bacterial contact angles: difficulties in defining hydrophobicity and surface Gibbs energy.

One of the principal techniques for evaluating the surface hydrophobicity of biological samples is contact angle. This method, applied readily to flat-smooth-inert surfaces, gives rise to an important debate when implemented with microbial lawns. After its initial description, in 1984, several authors have carried out modifications of the technique but the results obtained have not been previously judged.

In situ characterization of differences in the viscoelastic response of individual gram-negative and gram-positive bacterial cells.

We used a novel atomic force microscopy (AFM)-based technique to compare the local viscoelastic properties of individual gram-negative (Escherichia coli) and gram-positive (Bacillus subtilis) bacterial cells. We found that the viscoelastic properties of the bacterial cells are well described by a three-component mechanical model that combines an instantaneous elastic response and a delayed elastic response. These experiments have allowed us to investigate the relationship between the viscoelastic properties and the structure and composition of the cell envelope.

Surface viscoelasticity of individual gram-negative bacterial cells measured using atomic force microscopy.

The cell envelope of gram-negative bacteria is responsible for many important biological functions: it plays a structural role, it accommodates the selective transfer of material across the cell wall, it undergoes changes made necessary by growth and division, and it transfers information about the environment into the cell. Thus, an accurate quantification of cell mechanical properties is required not only to understand physiological processes but also to help elucidate the relationship between cell surface structure and function. We have used a novel, atomic force microscopy (AFM)-based approach to probe the mechanical properties of single bacterial cells by applying a constant compressive force to the cell under fluid conditions while measuring the time-dependent displacement (creep) of the AFM tip due to the viscoelastic properties of the cell.

Localized attraction correlates with bacterial adhesion to glass and metal oxide substrata.

Bacterial adhesion to surfaces does not always proceed according to theoretical expectations. Discrepancies are often attributed to surface heterogeneities that provide localized, favorable sites for bacterial attachment. The presence of these favorable deposition sites for bacteria, however, has never been directly measured.

Residence time, loading force, pH, and ionic strength affect adhesion forces between colloids and biopolymer-coated surfaces.

Exopolymers are thought to influence bacterial adhesion to surfaces, but the time-dependent nature of molecular-scale interactions of biopolymers with a surface are poorly understood. In this study, the adhesion forces between two proteins and a polysaccharide [Bovine serum albumin (BSA), lysozyme, or dextran] and colloids (uncoated or BSA-coated carboxylated latex microspheres) were analyzed using colloid probe atomic force microscopy (AFM). Increasing the residence time of an uncoated or BSA-coated microsphere on a surface consistently increased the adhesion force measured during retraction of the colloid from the surface, demonstrating the important contribution of polymer rearrangement to increased adhesion force.

Role of lactobacillus cell surface hydrophobicity as probed by AFM in adhesion to surfaces at low and high ionic strength.

The S-layer present at the outermost cell surface of some lactobacillus species is known to convey hydrophobicity to the lactobacillus cell surface. Yet, it is commonly found that adhesion of lactobacilli to solid substrata does not proceed according to expectations based on cell surface hydrophobicity. In this paper, the role of cell surface hydrophobicity of two lactobacillus strains with and without a surface layer protein (SLP) layer has been investigated with regard to their adhesion to hydrophobically or hydrophilically functionalized glass surfaces under well-defined flow conditions and in low and high ionic strength suspensions.

Dynamic cell surface hydrophobicity of Lactobacillus strains with and without surface layer proteins.

Variations in surface hydrophobicity of six Lactobacillus strains with and without an S-layer upon changes in ionic strength are derived from contact angle measurements with low- and high-ionic-strength aqueous solutions. Cell surface hydrophobicity changed in response to changes in ionic strength in three out of the six strains, offering these strains a versatile mechanism to adhere to different surfaces. The dynamic behavior of the cell surface hydrophobicity could be confirmed for two selected strains by measuring the interaction force between hydrophobic and hydrophilic tips with use of atomic force microscopy.

Comparison of atomic force microscopy interaction forces between bacteria and silicon nitride substrata for three commonly used immobilization methods.

Atomic force microscopy (AFM) has emerged as a powerful technique for mapping the surface morphology of biological specimens, including bacterial cells. Besides creating topographic images, AFM enables us to probe both physicochemical and mechanical properties of bacterial cell surfaces on a nanometer scale. For AFM, bacterial cells need to be firmly anchored to a substratum surface in order to withstand the friction forces from the silicon nitride tip.

Atomic force microscopic corroboration of bond aging for adhesion of Streptococcus thermophilus to solid substrata.

Initial bacterial adhesion is considered to be reversible, but over time the adhesive bond between a bacterium and a substratum surface may strengthen, turning the process into an irreversible state. Microbial desorption has been studied in situ in controlled flow devices as a function of the organisms resident time on the surface (J. Colloid Interface Sci.

Relations between macroscopic and microscopic adhesion of Streptococcus mitis strains to surfaces.

Application of physico-chemical models to describe bacterial adhesion to surfaces has hitherto only been partly successful due to the structural and chemical heterogeneities of bacterial surfaces, which remain largely unaccounted for in macroscopic physico-chemical characterizations of the cell surfaces. In this study, the authors attempted to correlate microscopic adhesion of a collection of nine Streptococcus mitis strains to the negatively charged, hydrophilic silicon nitride tip of an atomic force microscope (AFM) with macroscopic adhesion of the strains to a negatively charged, hydrophilic glass in a parallel-plate flow chamber. The repulsive force probed by AFM upon approach of the tip to a bacterial cell surface ranged from 1.

Softness of the bacterial cell wall of Streptococcus mitis as probed by microelectrophoresis.

Chemical and structural complexity of bacterial cell surfaces complicate accurate quantification of cell surfaces properties. The presence of fibrils, fimbriae or other surface appendages on bacterial cell surfaces largely influence those properties and would therefore play a major function in interfacial phenomena as aggregation and adhesion. The electrophoretic softness and fixed charge density in the polyelectrolyte layer of nine Streptococcus mitis strains, usually carrying long sparsely distributed fibrils, were determined by the soft particle analysis using measured electrophoretic mobilities as a function of the ionic strength.