Development and use of a parallel‐plate flow chamber for studying cellular adhesion to solid surfaces

Kooten, van Theo; Schakenraad, Jm; Mei, van der Henny C.; Busscher, Henk J.

doi:10.1002/jbm.820260604

Cited by 125 publications

(69 citation statements)

References 19 publications

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“…A parallel plate flow chamber permits quantitative reproducible adhesion measurements between yeast cells and a surface by applying a known, regulatable shear stress under conditions of laminar flow (40,64). This shear stress is felt as a shear force, which can detach the cells into the bulk medium or roll them along the surface.…”

Section: Flo8mentioning

confidence: 99%

EAP1 , a Candida albicans Gene Involved in Binding Human Epithelial Cells

2003

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Candida albicans adhesion to host tissues contributes to its virulence and adhesion to medical devices permits biofilm formation, but we know relatively little about the molecular mechanisms governing C. albicans adhesion to materials or mammalian cells. Saccharomyces cerevisiae provides an attractive model system for studying adhesion in yeast because of its well-characterized genetics and gene expression systems and the conservation of signal transduction pathways among the yeasts. In this study, we used a parallel plate flow chamber to screen and characterize attachment of a flo8⌬ S. cerevisiae strain expressing a C. albicans genomic library to a polystyrene surface. The gene EAP1 was isolated as a putative cell wall adhesin. Sequence analysis of EAP1 shows that it contains a signal peptide, a glycosylphosphatidylinositol anchor site, and possesses homology to many other yeast genes encoding cell wall proteins. In addition to increasing adhesion to polystyrene, heterologous expression of EAP1 in S. cerevisiae and autonomous expression of EAP1 in a C. albicans efg1 homozygous null mutant significantly enhanced attachment to HEK293 kidney epithelial cells. EAP1 expression also restored invasive growth to haploid flo8⌬ and flo11⌬ strains as well as filamentous growth to diploid flo8/flo8 and flo11/flo11 strains. Transcription of EAP1 in C. albicans is regulated by the transcription factor Efg1p, suggesting that EAP1 expression is activated by the cyclic AMP-dependent protein kinase pathway.

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Section: Flo8mentioning

confidence: 99%

EAP1 , a Candida albicans Gene Involved in Binding Human Epithelial Cells

2003

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“…Incubation and rinse (19,46), column flow (4,38), and flow chamber experiments (21,45) cannot be used to study bacteria on a submicrometer level. The technique of atomic force microscopy (AFM) is a powerful tool that has been used to study individual bacteria and bacterial adhesion, and researchers have recently examined polymer and extracellular polymeric substance attachment strength related to bacterial adhesion (7,41).…”

mentioning

confidence: 99%

Oriented Adhesion of Escherichia coli to Polystyrene Particles

Jones

Feick

Imoudu

et al. 2003

Appl Environ Microbiol

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The adhesion of nonflagellated Escherichia coli strain K-12 to polystyrene (PS) latex spheres or glass capillaries has been observed by using several techniques. Attention was focused on the orientation of the rod-shaped bacteria as they adhered to the surfaces in 100 mM phosphate-buffered saline. Data show that PS particles adhered to the ends of the bacteria more than 90% of the time. Moreover, the PS particles adhered to one end only, never to both. Similarly, for experiments with bacteria adhering to glass, the bacteria adhered on their ends. In order to determine whether the end of a bacterium had a different charge density from that of the middle, rotational electrophoresis experiments were used. These experiments indicated no measurable charge nonuniformity. In order to examine how strongly adhered the bacteria were to the PS particles, differential electrophoresis was used. Almost always, bacteria were found to be irreversibly adhered to the PS spheres. The cause of the oriented adhesion is not likely due to surface lipopolysaccharides (LPS), since the three strains of K-12 that were used, each having a different length of LPS, showed similar behavior. The results are discussed in terms of bacterial cell polarity. The data indicate that nanodomains on the bacterial ends are important for adhesion and that the time scale for irreversible adhesion is short.

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“…This variation is presumably due to various pharmacokinetic profiles. Systems such as CDFF (35) and flow chambers (47), for example, use a continuous flow of growth medium, while culture plate models (11) and chemostats (4) may have considerably longer antimicrobial retention times. The relatively large amplitude of the bacterial effects observed in this study may be attributable to the extended TR residence times due to the relatively slow flow rate of artificial saliva.…”

Section: Resultsmentioning

confidence: 99%

Effects of Triclosan-Containing Rinse on the Dynamics and Antimicrobial Susceptibility of In Vitro Plaque Ecosystems

McBain

Bartolo

Catrenich

et al. 2003

Antimicrob Agents Chemother

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Dental plaque microcosms were established under a feast-famine regimen within constant-depth film fermentors and exposed four times daily postfeeding to a triclosan (TR)-containing rinse (dentifrice) (TRD). This was diluted so that the antimicrobial content was 0.6 mg/ml. Microcosms were characterized by heterotrophic plate counts and PCR-denaturing gradient gel electrophoresis (DGGE) with primers specific for the V2-V3 region of the eubacterial 16S rRNA gene (rDNA). Dominant isolates and PCR amplicons were identified by partial sequencing of 16S rDNA. TRD caused considerable decreases in the counts of both gram-negative organisms and total anaerobic cells, transiently lowered the numbers of streptococci and actinomycetes, and markedly increased the proportion of lactobacilli. DGGE indicated the presence of putatively unculturable bacteria and showed that a Porphyromonas sp. and Selenomonas infelix had been inhibited by TRD. Pure culture studies of 10 oral bacteria (eight genera) showed that Neisseria subflava, Prevotella nigrescens, and Porphyromonas gingivalis were highly susceptible to TR, while the lactobacilli and streptococci were the least susceptible. Clonal expansion of the lactobacilli in the pulsed microcosm could be explained on the basis of TR activity. The mean MICs of TR, chlorhexidine, erythromycin, penicillin V, and vancomycin for the population before and after 5 days of exposure to TRD showed few significant changes. In conclusion, changes in plaque microcosm populations following repeated exposure to TRD showed inhibition of the most susceptible flora and clonal expansion of less susceptible species.

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Development and use of a parallel‐plate flow chamber for studying cellular adhesion to solid surfaces

Cited by 125 publications

References 19 publications

EAP1 , a Candida albicans Gene Involved in Binding Human Epithelial Cells

EAP1 , a Candida albicans Gene Involved in Binding Human Epithelial Cells

Oriented Adhesion of Escherichia coli to Polystyrene Particles

Effects of Triclosan-Containing Rinse on the Dynamics and Antimicrobial Susceptibility of In Vitro Plaque Ecosystems

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