Chlorhexidine-induced elastic and adhesive changes of Escherichia coli cells within a biofilm.

Chlorhexidine is a widely used, commercially available cationic antiseptic. Although its mechanism of action on planktonic bacteria has been well explored, far fewer studies have examined its interaction with an established biofilm. The physical effects of chlorhexidine on a biofilm are particularly unknown. Here, the authors report the first observations of chlorhexidine-induced elastic and adhesive changes to single cells within a biofilm. The elastic changes are consistent with the proposed mechanism of action of chlorhexidine. Atomic force microscopy and force spectroscopy techniques were used to determine spring constants and adhesion energy of the individual bacteria within an Escherichia coli biofilm. Medically relevant concentrations of chlorhexidine were tested, and cells exposed to 1% (w/v) and 0.1% more than doubled in stiffness, while those exposed to 0.01% showed no change in elasticity. Adhesion to the biofilm also increased with exposure to 1% chlorhexidine, but not for the lower concentrations tested. Given the prevalence of chlorhexidine in clinical and commercial applications, these results have important ramifications on biofilm removal techniques.

Characterizing pilus-mediated adhesion of biofilm-forming E. coli to chemically diverse surfaces using atomic force microscopy.

Biofilms are complex communities of microorganisms living together at an interface. Because biofilms are often associated with contamination and infection, it is critical to understand how bacterial cells adhere to surfaces in the early stages of biofilm formation. Even harmless commensal Escherichia coli naturally forms biofilms in the human digestive tract by adhering to epithelial cells, a trait that presents major concerns in the case of pathogenic E. coli strains. The laboratory strain E. coli ZK1056 provides an intriguing model system for pathogenic E. coli strains because it forms biofilms robustly on a wide range of surfaces.E. coli ZK1056 cells spontaneously form living biofilms on polylysine-coated AFM cantilevers, allowing us to measure quantitatively by AFM the adhesion between native biofilm cells and substrates of our choice. We use these biofilm-covered cantilevers to probe E. coli ZK1056 adhesion to five substrates with distinct and well-characterized surface chemistries, including fluorinated, amine-terminated, and PEG-like monolayers, as well as unmodified silicon wafer and mica. Notably, after only 0-10 s of contact time, the biofilms adhere strongly to fluorinated and amine-terminated monolayers as well as to mica and weakly to "antifouling" PEG monolayers, despite the wide variation in hydrophobicity and charge of these substrates. In each case the AFM retraction curves display distinct adhesion profiles in terms of both force and distance, highlighting the cells' ability to adapt their adhesive properties to disparate surfaces. Specific inhibition of the pilus protein FimH by a nonhydrolyzable mannose analogue leads to diminished adhesion in all cases, demonstrating the critical role of type I pili in adhesion by this strain to surfaces bearing widely different functional groups. The strong and adaptable binding of FimH to diverse surfaces has unexpected implications for the design of antifouling surfaces and antiadhesion therapies.

Friction dependence on growth conditions in epitaxial films.

In investigating the growth kinetics of epitaxial films in situ by force microscopy, we have observed several instances where the lateral force contrast on the growing monolayer exhibits a strong dependence on the driving force for growth (i.e., solute concentration). We present results for three epitaxial growth systems in aqueous solutions: CaSO(3) on CaCO(3), PbSO(4) on BaSO(4), and BaSO(3) on BaSO(4). In each system, material grown at higher solute concentrations exhibits a friction higher than that of material grown at lower concentrations. These observations suggest a link between defect density and friction contrast in growing epitaxial films. An additional time-dependent behavior is observed in the CaSO(3)/CaCO(3) system, indicating an annealing process.

Two-dimensional crystal growth from undersaturated solutions.

The solubility of a substance is commonly understood as the minimum concentration necessary for the condensation of a solid phase from solution. Here we report the nucleation and growth of ionic compounds from aqueous concentrations on the order of 0.1 times the solubility. The condensation is catalyzed by a foreign substrate, and the new phase grows as a crystalline monolayer. Undersaturated growth is observed only in cases where the dissolved compound is isomorphic with the substrate and the interaction strength between a dissolved-ion/substrate-ion pair exceeds that between the two dissolved ions. These results are consistent with a simple model in which favorable ion-surface interactions lead to ion enrichment and supersaturation in the two-dimensional interfacial zone.