Transmission electron microscopy of early microbial colonization of human enamel and root surfaces in vivo

Nyvad, Bente; Fejerskov, O.

doi:10.1111/j.1600-0722.1987.tb01845.x

Cited by 53 publications

(41 citation statements)

References 39 publications

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“…Although these numbers are 10-to 100-fold lower than total counts reported in previous studies (25), the increase in cell numbers between 4 and 8 h in the present study and in reference 24 was strikingly similar: 2-to 40-fold. Collectively, the relative amounts of colonization at 4 and 8 h and the relative ratios of actinomyces to strepto- cocci at 4 and 8 h, as well as the predominance of streptococci in early dental plaque, support and extend previous work (21)(22)(23)25).…”

Section: Discussionsupporting

confidence: 72%

Coaggregation-Mediated Interactions of Streptococci and Actinomyces Detected in Initial Human Dental Plaque

et al. 2003

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Streptococci and actinomyces that initiate colonization of the tooth surface frequently coaggregate with each other as well as with other oral bacteria. These observations have led to the hypothesis that interbacterial adhesion influences spatiotemporal development of plaque. To assess the role of such interactions in oral biofilm formation in vivo, antibodies directed against bacterial surface components that mediate coaggregation interactions were used as direct immunofluorescent probes in conjunction with laser confocal microscopy to determine the distribution and spatial arrangement of bacteria within intact human plaque formed on retrievable enamel chips. In intrageneric coaggregation, streptococci such as Streptococcus gordonii DL1 recognize receptor polysaccharides (RPS) borne on other streptococci such as Streptococcus oralis 34. To define potentially interactive subsets of streptococci in the developing plaque, an antibody against RPS (anti-RPS) was used together with an antibody against S. gordonii DL1 (anti-DL1). These antibodies reacted primarily with single cells in 4-h-old plaque and with mixed-species microcolonies in 8-h-old plaque. Anti-RPS-reactive bacteria frequently formed microcolonies with anti-DL1-reactive bacteria and with other bacteria distinguished by general nucleic acid stains. In intergeneric coaggregation between streptococci and actinomyces, type 2 fimbriae of actinomyces recognize RPS on the streptococci. Cells reactive with antibody against type 2 fimbriae of Actinomyces naeslundii T14V (anti-type-2) were much less frequent than either subset of streptococci. However, bacteria reactive with anti-type-2 were seen in intimate association with anti-RPS-reactive cells. These results are the first direct demonstration of coaggregation-mediated interactions during initial plaque accumulation in vivo. Further, these results demonstrate the spatiotemporal development and prevalence of mixed-species communities in early dental plaque.The human oral cavity harbors a complex microbial ecosystem characterized by spatiotemporal variability in species composition. Despite this variability, consensus exists that supra-and subgingival dental plaques develop according to reproducible patterns. Analyses of species composition in supragingival dental plaque have shown that the majority (47 to 90%) of cultivable bacteria are Streptococcus sanguinis (formerly S. sanguis [31]), Streptococcus oralis, and Streptococcus mitis (biovar 1) (24) and that one-third of the remaining bacteria are Actinomyces naeslundii (25). Scanning electron microscopy has shown isolated cells and clusters of cells after 4 h of accumulation, larger "microcolonies" after 8 h, and confluent monolayers after 12 h (22). Transmission electron microscopy has shown that some colonies consisted of gram-negative cells together with gram-positive cells; thus, multispecies colonies were unambiguously identifiable by 24 h (23). These and similar studies together form the basis for present understanding of community evolution in early sup...

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Section: Discussionsupporting

confidence: 72%

Coaggregation-Mediated Interactions of Streptococci and Actinomyces Detected in Initial Human Dental Plaque

et al. 2003

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“…Plaque biovars were seen to develop at exactly the same rate from individual to individual, reaching a plateau around 12 hours after stent insertion (Nyvad & Kilian, 1987). Electron microscopy confirmed a change in species composition over the next 12 hours with both Gram-positive and Gram-negative bacteria appearing, providing evidence for direct interaction between species in the biofilm (Nyvad & Fejerskov, 1987b). Further studies confirmed the importance of cell-to-cell recognition in early plaque development and examination of undisturbed plaque.…”

Section: Bacterial Interactions During Biofilm Developmentsupporting

confidence: 56%

“…The proximity of phenotypes allows for their interactions and influences their ability to survive within the biofilm. Among the early studies of spatial relationships in plaque biofilm formation are the studies by Nyvad and colleagues (Nyvad, 1993;Nyvad & Fejerskov, 1987a;Nyvad & Fejerskov, 1987b;Nyvad & Killian, 1987). Using a stent that holds enamel pieces (commonly used in supragingival oral film investigations), they placed it in the oral cavity and monitored the formation of plaque biofilms.…”

Section: Bacterial Interactions During Biofilm Developmentmentioning

confidence: 99%

The Microbial Aetiology of Periodontal Diseases

Africa¹

2012

Periodontal Diseases - A Clinician's Guide

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“…With increasing age of the biofilm, microcolonies of A. naeslundii and other non-streptococci were seen to extend perpendicularly from the supporting surface as 'chimney' structures and palisades like those observed by electron microscopy of multi-layered dental plaque (Listgarten et al, 1975;Nyvad & Fejerskov, 1987b;Rosan et al, 1976). The morphogenesis of these particular structures can only be speculated on.…”

Section: Discussionmentioning

confidence: 99%

“…For the first time, our study provides a presentation of the spatio-temporal organization of A. naeslundii in relation to other bacteria in initial multi-layer dental biofilms formed in vivo up to 48 h and definitively demonstrates that A. naeslundii preferentially occupies the inner part of early multilayered biofilms. We therefore infer that the densely packed, pleomorphic Gram-positive bacteria with thick cell walls previously observed close to the enamel surface in electron micrographs (Listgarten et al, 1975;Nyvad & Fejerskov, 1987b;Schroeder & De Boever, 1970) are A. naeslundii.…”

Section: Discussionmentioning

confidence: 99%

Actinomyces naeslundii in initial dental biofilm formation

et al. 2009

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The combined use of confocal laser scanning microscopy (CLSM) and fluorescent in situ hybridization (FISH) offers new opportunities for analysis of the spatial relationships and temporal changes of specific members of the microbiota of intact dental biofilms. The purpose of this study was to analyse the patterns of colonization and population dynamics of Actinomyces naeslundii compared to streptococci and other bacteria during the initial 48 h of biofilm formation in the oral cavity. Biofilms developed on standardized glass slabs mounted in intra-oral appliances worn by ten individuals for 6, 12, 24 and 48 h. The biofilms were subsequently labelled with probes against A. naeslundii (ACT476), streptococci (STR405) or all bacteria (EUB338), and were analysed by CLSM. Labelled bacteria were quantified by stereological tools. The results showed a notable increase in the number of streptococci and A. naeslundii over time, with a tendency towards a slower growth rate for A. naeslundii compared with streptococci. A. naeslundii was located mainly in the inner part of the multilayered biofilm, indicating that it is one of the species that attaches directly to the acquired pellicle. The participation of A. naeslundii in the initial stages of dental biofilm formation may have important ecological consequences. INTRODUCTIONDental biofilm is an archetypal example of a complex biofilm (Costerton et al., 1999;Davies, 2003;DuPont, 1997). Biofilm formation on tooth surfaces follows the same basic rules as biofilm formation elsewhere in nature. Dental biofilms develop readily because of the optimal temperature, the rich nutrient supply in the oral cavity, and the hard non-shedding surface. They are easily accessible for experimentation using intra-oral devices (Auschill et al., 2004;Kilian et al., 1979;Nyvad & Fejerskov, 1987a;Palmer et al., 2003), and therefore dental biofilms can be used to demonstrate colonization phenomena and ecological principles of universal interest. Concurrent with the increasing recognition of the significance of biofilms in infectious diseases, the development of techniques such as confocal laser scanning microscopy (CLSM), fluorescence in situ hybridization (FISH) and immunofluorescence has enabled visualization of bacteria in their natural undisturbed environment. This offers a substantial improvement upon previous microbiological studies of bacteria grown in planktonic settings (Anwar et al., 1992;Davies, 2003).Previous studies of dental biofilm that took advantage of these methods mainly focused on streptococci Dige et al., 2007; Hannig et al., 2007;Palmer et al., 2003) because culture-based studies suggested that this group of bacteria is prominent during the initial stages of biofilm formation on teeth (Li et al., 2004;Nyvad & Kilian, 1987. However, other genera such as Actinomyces are also among the earliest colonizers of dental surfaces and may constitute up to 27 % of the pioneer bacteria (Kilian et al., 1979;Li et al., 2004;Nyvad & Kilian, 1987). Several culturebased studies indicated that Ac...

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Transmission electron microscopy of early microbial colonization of human enamel and root surfaces in vivo

Cited by 53 publications

References 39 publications

Coaggregation-Mediated Interactions of Streptococci and Actinomyces Detected in Initial Human Dental Plaque

Coaggregation-Mediated Interactions of Streptococci and Actinomyces Detected in Initial Human Dental Plaque

The Microbial Aetiology of Periodontal Diseases

Actinomyces naeslundii in initial dental biofilm formation

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