Quantitative Method for Determining the Lateral Strength of Bacterial Adhesion and Application for Characterizing Adhesion Kinetics

Deupree, Susan M.

doi:10.1021/la800417g

Cited by 22 publications

(31 citation statements)

References 42 publications

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“…Centrifugation in essence involves compacting bacteria into a pellet, causing collisions against each other that result in shear forces on the bacterial cell surface, which may easily lead to cell surface damage with a potential effect on the outcome of surface-sensitive experiments. Many experimental protocols choose high centrifugation speeds to collect as many bacteria as possible while assuming that it does not cause any bacterial cell damage or cell death (4,10). However, cell surface damage during bacterial centrifugation at 15,000 ϫ g was demonstrated to cause significant reductions in Escherichia coli viability compared to centrifugation at 5,000 ϫ g, while little effect was detected on the viability of Psychrobacter sp.…”

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Bacterial Cell Surface Damage Due to Centrifugal Compaction

Peterson

Sharma

Mei

et al. 2012

Appl Environ Microbiol

121

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Centrifugal damage has been known to alter bacterial cell surface properties and interior structures, including DNA. Very few studies exist on bacterial damage caused by centrifugation because of the difficulty in relating centrifugation speed and container geometry to the damage caused. Here, we provide a simple, versatile method of analysis for describing the compaction of bacteria during centrifugation based on a proposed centrifugation coefficient, C. Values of C can be related to different bacterial cell surface properties. Changing the geometry of the centrifugation container or centrifugation speeds changed the value of C significantly. Initial deposition rates of Staphylococcus aureus ATCC 12600 to a glass surface decayed exponentially from 4,217 to 1,478 cm ؊2 s ؊1 with increasing C, while the proportion of staphylococci with a zeta potential of around ؊15 mV decreased from 97 to 58%. These surface-sensitive parameters were used independently to derive a critical centrifugation coefficient (0.040), above which centrifugation was considered to impact the outcome of surface-sensitive experiments due to cell surface damage. The critical centrifugation coefficient could successfully predict staphylococcal cell surface damage, i.e., a significant change in initial deposition rate or zeta potential distribution, in 84% of all cases included here, whereas the centrifugation speed could predict damage in only 58% of all cases. Moreover, controlling the centrifugation coefficient within narrow limits over a series of experiments yielded 43% smaller standard deviations in initial staphylococcal deposition rates than with centrifugation at fixed speeds for replicate experiments. C entrifugation is a common laboratory practice used for harvesting planktonic bacteria. For harvesting, a wide variety of forces (roughly ranging from 1,000 to 12,000 ϫ g) (1, 10-12) is used without mentioning a reason for a particular choice. Centrifugation in essence involves compacting bacteria into a pellet, causing collisions against each other that result in shear forces on the bacterial cell surface, which may easily lead to cell surface damage with a potential effect on the outcome of surface-sensitive experiments. Many experimental protocols choose high centrifugation speeds to collect as many bacteria as possible while assuming that it does not cause any bacterial cell damage or cell death (4, 10). However, cell surface damage during bacterial centrifugation at 15,000 ϫ g was demonstrated to cause significant reductions in Escherichia coli viability compared to centrifugation at 5,000 ϫ g, while little effect was detected on the viability of Psychrobacter sp. strain SW8 or Staphylococcus epidermidis (9). Moreover, bacterial cell surface damage can be expected to affect surface-sensitive phenomena, such as their adhesion to surfaces. Chemical and physical surface damage to Pseudomonas aeruginosa, for instance, yielded lower initial deposition rates to substratum surfaces than were observed for undamaged bacteria (2). In general, ...

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confidence: 99%

Bacterial Cell Surface Damage Due to Centrifugal Compaction

Peterson

Sharma

Mei

et al. 2012

Appl Environ Microbiol

121

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“…In previous works2829, the AFM tip was used to repeatedly scan over the cells until they were detached. The disadvantage of this technique is that the adhesion properties of the cells might have been altered while being scanned.…”

Section: Resultsmentioning

confidence: 99%

“…The technique used in this study was a little bit different with the one described in the literature2829. Two types of cantilevers were used in these experiments.…”

Section: Materials and Modelmentioning

confidence: 99%

“…In order to quantitatively measure the shear force required to detach an individual cell, Deupree and Schoenfisch28 developed a novel method based on the total compression of the AFM cantilever during cell detachment events. This method has been then modified to account for the bending of cantilever due to the counterforce from the cell during detachment29.…”

Section: Materials and Modelmentioning

confidence: 99%

“…The experiments will be conducted by applying a shear force on the cells until they are detached. Details of this technique were already described in the literature2829 (see Fig. 1c).…”

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confidence: 99%

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Investigation of Cell-Substrate Adhesion Properties of Living Chondrocyte by Measuring Adhesive Shear Force and Detachment Using AFM and Inverse FEA

Nguyen

2016

Sci Rep

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It is well-known that cell adhesion is important in many biological processes such as cell migration and proliferation. A better understanding of the cell adhesion process will shed insight into these cellular biological responses as well as cell adhesion-related diseases treatment. However, there is little research which has attempted to investigate the process of cell adhesion and its mechanism. Thus, this paper aims to study the time-dependent adhesion properties of single living chondrocytes using an advanced coupled experimental-numerical approach. Atomic Force Microscopy (AFM) tips will be used to apply lateral forces to detach chondrocytes that are seeded for three different periods. An advanced Finite Element Analysis (FEA) model combining porohyperelastic (PHE) constitutive model and cohesive zone formulation is developed to explore the mechanism of adhesion. The results revealed that the cells can resist normal traction better than tangential traction in the beginning of adhesion. This is when the cell adhesion molecules establish early attachment to the substrates. After that when the cells are spreading, stress fiber bundles generate tangential traction on the substrate to form strong adhesion. Both simulation and experimental results agree well with each other, providing a powerful tool to study the cellular adhesion process.

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Morphological analysis of the antimicrobial action of nitric oxide on Gram-negative pathogens using atomic force microscopy

Deupree

Schoenfisch

2009

Acta Biomaterialia

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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.

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Quantitative Method for Determining the Lateral Strength of Bacterial Adhesion and Application for Characterizing Adhesion Kinetics

Cited by 22 publications

References 42 publications

Bacterial Cell Surface Damage Due to Centrifugal Compaction

Bacterial Cell Surface Damage Due to Centrifugal Compaction

Investigation of Cell-Substrate Adhesion Properties of Living Chondrocyte by Measuring Adhesive Shear Force and Detachment Using AFM and Inverse FEA

Morphological analysis of the antimicrobial action of nitric oxide on Gram-negative pathogens using atomic force microscopy

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