Utilization of a Continuous Streptococcal Surface to Measure Interbacterial Adherence in vitro and in vivo

Liljemark, W. F.; Bloomquist, C. G.; Coulter, M. C.; Fenner, Laurie J.; Skopek, R. J.; Schachtele, Charles F.

doi:10.1177/00220345880670120301

Cited by 27 publications

(29 citation statements)

References 33 publications

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“…That technology allows for more precise measurement of plaque accumulation and composition. It also facilitates collection of site-specific samples (Liljemark et al, 1988;Skopek et al, 1993). That may be very important in view of the finding that salivary protein binding by bacteria differs between oral sites (Rudney et al, 1995).…”

Section: (C) Short-term Outcome Variablesmentioning

confidence: 99%

Does Variability in Salivary Protein Concentrations Influence Oral Microbial Ecology and Oral Health?

Rudney

1995

Critical Reviews in Oral Biology & Medicine

114

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Salivary protein interactions with oral microbes in vitro include aggregation, adherence, cell-killing, inhibition of metabolism, and nutrition. Such interactions might be expected to influence oral ecology. However, inconsistent results have been obtained from in vivo tests of the hypothesis that quantitative variation in salivary protein concentrations will affect oral disease prevalence. Results may have been influenced by choices made during study design, including saliva source, stimulation status, control for flow rate, and assay methods. Salivary protein concentrations also may be subject to circadian variation. Values for saliva collected at the same time of day tend to remain consistent within subjects, but events such as stress, inflammation, infection, menstruation, or pregnancy may induce short-term changes. Long-term factors such as aging, systemic disease, or medication likewise may influence salivary protein concentrations. Such sources of variation may increase the sample size needed to find statistically significant differences. Clinical studies also must consider factors such as human population variation, strain and species differences in protein-microbe interactions, protein polymorphism, and synergistic or antagonistic interaction between proteins. Salivary proteins may form heterotypic complexes with unique effects, and different proteins may exert redundant effects. Patterns of protein-microbe interaction also may differ between oral sites. Future clinical studies must take those factors into account. Promising approaches might involve meta-analysis or multi-center studies, retrospective and prospective longitudinal designs, short-term measurement of salivary protein effects, and consideration of individual variation in multiple protein effects such as aggregation, adherence, and cell-killing.

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Section: (C) Short-term Outcome Variablesmentioning

confidence: 99%

Does Variability in Salivary Protein Concentrations Influence Oral Microbial Ecology and Oral Health?

Rudney

1995

Critical Reviews in Oral Biology & Medicine

114

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“…Scanning electron microscopy has enabled estimations of the space occupied on the enamel by streptococcal cells, and these data were expressed as cells per surface area. Using scanning electron microscopy, we found that a mean of 1 to 2 million approximately 1-m 3 streptococci per mm 2 formed a continuous layer when linked to an enamel surface via a biological adhesive (25). Therefore, a population density of 2.5 ϫ 10 5 to 6.3 ϫ 10 5 1-m 3 cells per mm 2 would cover between 12 and 32% of the salivary pellicle with streptococci, assuming an even 1-cell-thick layer.…”

Section: Estimation Of Saturation Of Salivary-pellicle Binding Sitesmentioning

confidence: 99%

“…However, the numbers of bacteria saturating either saliva-coated enamel or a continuous layer of bacteria were too low to explain the large, rapid increase in plaque biomass that occurs in vivo by adherence alone (33). Therefore, an original model system was used to quantitatively measure plaque accumulation in the mouth on a measured area of a tooth surface (24,25). Thus, this study was designed to enumerate the in situ adherence, accumulation, and cell division of plaque bacteria and to evaluate the contributions of these mechanisms to the large biomass increase observed during the first 24-h period of developing dental plaque.…”

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confidence: 99%

Adherence, accumulation, and cell division of a natural adherent bacterial population

1996

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Oral Microbiol. Immunol. 8:5-15, 1993). Radiolabeled nucleoside incorporation was used to measure DNA synthesis concurrent with the taking of standard viable cell counts of the plaque samples. Results showed that in vivo plaque formation began with the rapid adherence of bacteria until ca. 12 to 32% of the enamel's salivary pellicle was saturated (ca. 2.5 ؋ 10 5 to 6.3 ؋ 10 5 cells per mm 2 ). The pioneer adherent species were predominantly those of the ''sanguis streptococci.'' At the above-noted density, the bacteria present on the salivary pellicle incorporated low levels of radiolabeled nucleoside per viable cell. As bacterial numbers reached densities between 8.0 ؋ 10 5 and 2.0 ؋ 10 6 cells per mm 2 , there was a small increase in the incorporation of radiolabeled nucleosides per cell. At 2.5 ؋ 10 6 to 4.0 ؋ 10 6 cells per mm 2 of enamel surface, there was a marked increase in the incorporation of radiolabeled nucleosides per cell which appeared to be cell-density dependent. The predominant species group in developing dental plaque films during densitydependent growth was the sanguis streptococci; however, most other species present showed similar patterns of increased DNA synthesis as the density noted above approached 2.5 ؋ 10 6 to 4.0 ؋ 10 6 cells per mm .The accumulation of bacterial dental plaques on tooth surfaces in humans is a common but complex process. Bacterial plaque is positively associated with the formation of dental caries and periodontal disease. Large expenditures of time and money by the research community have been and are being made to study dental plaque formation in order to understand this complex biofilm. In vitro studies have found that the initial steps in plaque formation occur via specific adherence mechanisms. A myriad of bacterial adhesins and salivary pellicle receptors have been studied for potential relevant adherence mechanisms (recently reviewed by Kolenbrander and London [21]). In most of these systems, e.g., adherence assays, bacteria exhibit saturation kinetics until a 0.1 to 30% surface coverage of the experimental pellicle-coated hydroxyapatite surface occurs (2,8,14,15,27,28). Adherence among bacteria has also been studied by a number of various in vitro assays (7,16,36,38,39). Recently, a preformed plaque model consisting of a continuous layer of different bacterial species formed on enamel or plastic surfaces (with saliva coating) was used as the substratum. These in vitro studies of interbacterial adherence have also determined saturation kinetics using several different species of indigenous oral bacteria as the continuous layer (38). However, the numbers of bacteria saturating either saliva-coated enamel or a continuous layer of bacteria were too low to explain the large, rapid increase in plaque biomass that occurs in vivo by adherence alone (33). Therefore, an original model system was used to quantitatively measure plaque accumulation in the mouth on a measured area of a tooth surface (24, 25). Thus, this study was designed to enumerate the in situ adherenc...

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“…Certain bacteria may adhere preferentially to preformed plaque rather than directly to the tooth surface (5). Finally, coadhesion between interdependent species in dental plaque may promote synergistic physiological relations (7,17).…”

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confidence: 99%

“…Thus, several researchers have recently proposed studying coadhesion of one strain, free in suspension, to a preformed layer of the second strain adhering on a given surface. For example, Liljemark et al used a continuous layer of Streptococcus sanguis cells affixed to plastic and enamel surfaces with a commercial adhesive (Cell-TAK) to study coadhesion with Haemophilus parainfluenzae and Streptococcus sobrinus (17). Ellen and coworkers have studied attachment of Bacteroides gingivalis to preformed layers of Actinomyces viscosus strains on salivacoated hydroxyapatite beads (7,27).…”

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Adhesion of Actinomyces viscosus to Porphyromonas (Bacteroides) gingivalis-coated hexadecane droplets

1991

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Interbacterial adhesion (coadhesion) is considered a major determinant of dental plaque ecology. In this report, we studied several aspects of the adhesion of Porphyromonas (Bacteroides) gingivalis to hexadecane in order to use the liquid hydrocarbon as a convenient substratum for coadhesion assays. Washed suspensions of hydrophobic P. gingivalis 2561 cells were vortexed with hexadecane to yield highly stable cell-coated droplets. Kinetics of coadhesion between Actinomyces viscosus cells and P. gingivalis-coated hexadecane droplets (PCHD) was subsequently studied. Aliquots of PCHD were added to A. viscosus suspensions, and the mixtures were gently rotated. Avid adhesion of A. viscosus cells to the immobilized P. gingivalis layer could be readily measured by the decrease in turbidity in the aqueous phase, following phase separation. Despite the ability of A. viscosus cells to adsorb to hexadecane following vigorous mixing, gentle mixing did not appreciably promote adhesion to bare hexadecane. Moreover, extensive microscopic examinations revealed that A. viscosus cells adhered exclusively to the bound P. gingivalis cells rather than to exposed areas of hexadecane. Coadhesion of A. viscosus to the PCHD appeared to follow first-order kinetics, attaining 80% levels within 30 min. Electron micrographs revealed A. viscosus cells adhering to the P. gingivalis cell layer adsorbed at the hexadecane-water interface. Interestingly, P. gingivalis cells did not appear to penetrate the hexadecane. A. viscosus mutants lacking type 1 or type 2 fimbriae or both were still able to bind to the PCHD. No obvious correlation was observed between relative hydrophobicity of A. viscosus strains and their binding to PCHD. However, defatted bovine serum albumin, an inhibitor of hydrophobic interactions, was the most potent inhibitor among those tested. The data suggest that this approach provides a simple, quantitative technique for studying kinetics of bacterial coadhesion which is amenable to both light and electron microscopic observation.

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Utilization of a Continuous Streptococcal Surface to Measure Interbacterial Adherence in vitro and in vivo

Cited by 27 publications

References 33 publications

Does Variability in Salivary Protein Concentrations Influence Oral Microbial Ecology and Oral Health?

Does Variability in Salivary Protein Concentrations Influence Oral Microbial Ecology and Oral Health?

Adherence, accumulation, and cell division of a natural adherent bacterial population

Adhesion of Actinomyces viscosus to Porphyromonas (Bacteroides) gingivalis-coated hexadecane droplets

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