Atomic force microscopy (AFM) was used to study the morphological changes of two Gram-negative pathogens, Pseudomonas aeruginosa and Escherichia coli, after exposure to nitric oxide (NO). The time-dependent effects of NO released from a xerogel coating and the concentration-dependent effects rendered by a small-molecule that releases NO in a bolus were examined and compared. Bacteria exhibited irregular and degraded exteriors. With NO-releasing surfaces, an increase in surface debris and disorganized adhesion patterns were observed compared to controls. Analysis of cell surface topography revealed that increasing membrane roughness correlated with higher doses of NO. At a lower total dose, NO delivered via a bolus resulted in greater membrane roughness than NO released from a surface via a sustained flux. At sub-inhibitory levels, treatment with amoxicillin, an antibiotic known to compromise the integrity of the cell wall, led to morphologies resembling those resulting from NO treatment. Our observations indicate that cell envelope deterioration is a visible consequence of NO-exposure for both Gram-negative species studied.
The synergistic activity between nitric oxide (NO) released from diazeniumdiolate-modified proline (PROLI/NO) and silver (I) sulfadiazine (AgSD) was evaluated against Escherichia coli, Enterococcus faecalis, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Staphylococcus epidermidis using a modified broth microdilution technique and a checkerboardtype assay. The combination of NO and AgSD was defined as synergistic when the fractional bactericidal concentration (FBC) was calculated to be <0.5 Gram-negative species were generally more susceptible to the individual antimicrobial agents than the Gram-positive bacteria. The in vitro synergistic activity of AgSD and NO observed against a range of pathogens strongly supports future investigation of this therapeutic combination, particularly for its potential use in the treatment of chronic and burn wounds.
A quantitative method for measuring the shear force required to detach individual adhered bacteria using atomic force microscopy (AFM) was developed. By determining the total compression of the cantilever during cell detachment events, a more accurate means of calculating the applied lateral force necessary to remove individual cells was achieved compared to previous methods. In addition, a tunable assay for monitoring the dynamics of Pseudomonas aeruginosa and Staphylococcus aureus adhesion strength was employed. The accumulation of force measurements over time allowed for the characterization of adhesion strength kinetics. P. aeruginosa reinforced its adhesion to the surface at a rate 7-fold faster than for S. aureus; the average adhesion strength of P. aeruginosa was larger than that of S. aureus at corresponding time points. Adhered cells of the same species and strain demonstrated a range of adhesion forces that broadened with time, indicating that the change in adhesion strength does not proceed uniformly.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.