A Quantitative Comparison of Selected Bacteria in Human Carious Dentine by Microscopic Counts

Ozaki, Kazumi; Matsuo, Takashi; Nakae, Hajime; Noiri, Yuichiro; Yoshiyama, Masahiro; Ebisu, Shigeyuki

doi:10.1159/000261635

Cited by 37 publications

(39 citation statements)

References 9 publications

(10 reference statements)

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“…and the total anaerobic load in carious dentine following bacterial enumeration by real-time PCR is in agreement with the previous observations. The reported inability to cultivate P. gingivalis or P. endodontalis from carious dentine is pertinent to this argument (19), even though the latter species can be identified in dentine caries by immunohistological staining with speciesspecific antisera (24). Both of these species were readily detected by real-time PCR in 52 and 60% of samples, respectively.…”

Section: Discussionmentioning

confidence: 94%

Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

et al. 2002

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The bacteria found in carious dentine were correlated with the tissue response of the dental pulps of 65 teeth extracted from patients with advanced caries and pulpitis. Standardized homogenates of carious dentine were plated onto selective and nonselective media under anaerobic and microaerophilic conditions. In addition, real-time PCR was used to quantify the recovery of anaerobic bacteria. Primers and fluorogenic probes were designed to detect the total anaerobic microbial load, the genera Prevotella and Fusobacterium, and the species Prevotella melaninogenica, Porphyromonas endodontalis, Porphyromonas gingivalis, and Micromonas (formerly Peptostreptococcus) micros. The pulpal pathology was categorized according to the cellular response and degenerative changes. Analysis of cultured bacteria showed a predominance of gram-positive microorganisms, particularly lactobacilli. Gram-negative bacteria were also present in significant numbers with Prevotella spp., the most numerous anaerobic group cultured. Real-time PCR analysis indicated a greater microbial load than that determined by colony counting. The total number of anaerobes detected was 41-fold greater by real-time PCR than by colony counting, while the numbers of Prevotella and Fusobacterium spp. detected were 82-and 2.4-fold greater by real-time PCR than by colony counting, respectively. Real-time PCR also identified M. micros, P. endodontalis, and P. gingivalis in 71, 60, and 52% of carious samples, respectively. Correlation matrices of the real-time PCR data revealed significant positive associations between M. micros and P. endodontalis detection and inflammatory degeneration of pulpal tissues. These anaerobes have been strongly implicated in endodontic infections that occur as sequelae to carious pulpitis. Accordingly, the data suggest that the presence of high levels of these bacteria in carious lesions may be indicative of irreversible pulpal pathology.The microbial populations involved in dental caries are known to be highly complex and variable and have not yet been fully identified, although key organisms are generally recognized to be associated with disease progression. The bacteria involved in caries initiation and early caries development, particularly the mutans group streptococci and lactobacilli, have been well documented (34). As the lesion progresses, there is a transition from predominantly facultative gram-positive bacteria in early caries to anaerobic gram-positive rods and cocci and gram-negative rods in deep carious lesions (15). Previous research has associated the presence of anaerobic gram-negative rods, such as Fusobacterium, Prevotella, and Porphyromonas, with symptomatic teeth (11, 19), infected pulps (38), and periapical abscesses (29, 35), whereas anaerobic gram-positive cocci, such as peptostreptococci, have been associated with apical infections (10).While it is well recognized that bacteria and their products play a major role in dental caries and associated pulpal inflammation (5), attempts to correlate clinical signs an...

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Section: Discussionmentioning

confidence: 94%

Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

et al. 2002

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“…The occurrence of coadhesive interactions in vivo might explain why typically a limited number of species are found in an infected root canal. The presence of gram-negative bacteria in infected dentin (3,11,15,34) and within root canals with intact dentin walls (28,40,41,42,43) suggests that these bacteria can penetrate dentin in vivo. Associations between gram-negative bacteria and other bacteria that are capable of dentinal tubule invasion may thus allow invasion by the gram-negative organisms.…”

Section: Discussionmentioning

confidence: 99%

Coinvasion of Dentinal Tubules by Porphyromonas gingivalis and Streptococcus gordonii Depends upon Binding Specificity of Streptococcal Antigen I/II Adhesin

et al. 2000

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Cell wall-anchored polypeptides of the antigen I/II family are produced by many species of oral streptococci. These proteins mediate adhesion of streptococci to salivary glycoproteins and to other oral microorganisms and promote binding of cells to collagen type I and invasion of dentinal tubules. Since infections of the root canal system have a mixed anaerobic bacterial etiology, we investigated the hypothesis that coadhesion of anaerobic bacteria with streptococci may facilitate invasive endodontic disease. Porphyromonas gingivalis ATCC 33277 cells were able to invade dentinal tubules when cocultured with Streptococcus gordonii DL1 (Challis) but not when cocultured with Streptococcus mutans NG8. An isogenic noninvasive mutant of S. gordonii, with production of SspA and SspB (antigen I/II family) polypeptides abrogated, was deficient in binding to collagen and had a 40% reduced ability to support adhesion of P. gingivalis. Heterologous expression of the S. mutans SpaP (antigen I/II) protein in this mutant restored collagen binding and tubule invasion but not adhesion to P. gingivalis or the ability to promote P. gingivalis coinvasion of dentin. An isogenic afimbrial mutant of P. gingivalis had 50% reduced binding to S. gordonii cells but was unaffected in the ability to coinvade dentinal tubules with S. gordonii wild-type cells. Expression of the S. gordonii SspA or SspB polypeptide on the surface of Lactococcus lactis cells endowed these bacteria with the abilities to bind P. gingivalis, penetrate dentinal tubules, and promote P. gingivalis coinvasion of dentin. The results demonstrate that collagen-binding and P. gingivalis-binding properties of antigen I/II polypeptides are discrete functions. Specificity of antigen I/II polypeptide recognition accounts for the ability of P. gingivalis to coinvade dentinal tubules with S. gordonii but not with S. mutans. This provides evidence that the specificity of interbacterial coadhesion may influence directly the etiology of pulpal and periapical diseases.

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“…Further, considerable evidence also links Actinomyces spp. with caries, especially root surface caries [Ozaki et al, 1994;Schüpbach et al, 1996]. However, limited epidemiological data are available on the role of Actinomyces spp.…”

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confidence: 99%

<i>Actinomyces</i> spp. in Supragingival Plaque of Ethnic Chinese Preschool Children with and without Active Dental Caries

Tang

Yip²,

Samaranayake

et al. 2003

Caries Res

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Very limited molecular epidemiological data are available on the role of Actinomyces spp. in the pathogenesis of caries in the primary dentition. Therefore, we investigated their distribution in supragingival plaque of ethnic Chinese preschool children from Singapore and Hong Kong, either with or without active caries. Plaque samples were taken from intact interproximal enamel areas using dental floss. Bacterial genomic DNA of each sample was extracted and variable regions of 16S ribosomal DNA amplified and labelled with digoxigenin. Oligonucleotide probes specific for Actinomyces bovis, Actinomyces gerencseriae, Actinomyces israelii, Actinomyces meyeri, Actinomyces odontolyticus, catalase-negative Actinomyces naeslundii (genospecies 1 and 2) and catalase-positive Actinomyces naeslundii genospecies 2 (previously Actinomyces viscosus serotype II) were used to detect these species using Southern hybridization with a Minislot and Miniblotter system. A. odontolyticus, A. gerencseriae and A. meyeri were detected with similar frequency in both Singapore and Hong Kong samples or in those with and without active caries. However, the prevalence of A. naeslundii was significantly different in the two locales (p < 0.05). A. odontolyticus (88.7%), A. gerencseriae (56.6%) and A. naeslundii (50.9%) were detected in a majority of the samples and the positive hybridization signals of A. gerencseriae in the caries-active group were stronger than from the caries-free group. A. bovis and A. israelii were undetectable in any of the samples. These data imply that A. odontolyticus, A. naeslundii and A. gerencseriae may play an important role in supragingival plaque formation on primary teeth in ethnic Chinese, with others such as A. meyeri contributing.

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A Quantitative Comparison of Selected Bacteria in Human Carious Dentine by Microscopic Counts

Cited by 37 publications

References 9 publications

Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

Coinvasion of Dentinal Tubules by Porphyromonas gingivalis and Streptococcus gordonii Depends upon Binding Specificity of Streptococcal Antigen I/II Adhesin

<i>Actinomyces</i> spp. in Supragingival Plaque of Ethnic Chinese Preschool Children with and without Active Dental Caries

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