Intracellular
            <i>Actinobacillus actinomycetemcomitans</i>
            and
            <i>Porphyromonas gingivalis</i>
            in Buccal Epithelial Cells Collected from Human Subjects

Rudney, Joel D.; Chen, Ruoqiong; Sedgewick, Gerald

doi:10.1128/iai.69.4.2700-2707.2001

Cited by 158 publications

(167 citation statements)

References 43 publications

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“…Yet, the complex relationship between the subgingival microbiota and the epithelium of the gingival crevice in such infections has only been partially determined. A great deal of evidence has accumulated which corroborates the ability of putative periodontal pathogens, such as Prevotella intermedia (Dorn et al, 1998), Actinobacillus actinomycetemcomitans (FivesTaylor et al, 1995;Meyer et al, 1996;Rudney et al, 2001), Porphyromonas gingivalis (Sandros et al, 1994;Lamont et al, 1995;Madianos et al, 1996), Fusobacterium nucleatum (Han et al, 2000), and several species of oral spirochaetes (Peters et al, 1999), to adhere to and invade oral epithelial tissues and cells. However, most of these studies were performed in vitro, using few reference bacterial strains, and oral epithelial cell lines, primary epithelial cell cultures or cells from the buccal mucosa.…”

Section: Introductionmentioning

confidence: 96%

Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions

Colombo¹,

Silva²,

Haffajee

et al. 2006

Journal of Medical Microbiology

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Bacterial invasion of host epithelial cells plays an important role in the pathogenesis of periodontal diseases; however, the interactions between subgingival species and the gingival crevice cells are not fully understood. This study determined the prevalence of a group of oral bacterial species on or in epithelial cells derived from periodontal pockets and the gingival crevice of subjects with periodontitis. Samples of epithelial cells were obtained from 120 sites with periodontal pockets ¢4 mm and 92 periodontally healthy sites from 49 patients (mean age 46?3±1?4 years; 43 % males) with chronic periodontitis. Bacteria in or on epithelial cells were separated from unattached bacteria by Percoll density-gradient centrifugation. The presence and levels of 33 oral species were determined in epithelial cell samples by whole genomic DNA probes and the checkerboard method. The most frequently detected species were Porphyromonas gingivalis (42 %), Treponema denticola (38 %), Prevotella intermedia (37 %), Streptococcus intermedius (36 %), Campylobacter rectus (35 %), Streptococcus sanguinis (35 %) and Streptococcus oralis (34 %). Species of Actinomyces were found in low prevalence and levels. The data indicated that there were more micro-organisms on or in epithelial cells obtained from periodontal pockets than from healthy sulci; however, no significant differences regarding the percentage and level of any specific species were found between these sites. Veillonella parvula, S. oralis, Streptococcus gordonii and Streptococcus mitis tended to be more prevalent in sites without disease. These findings demonstrated that a wide range of oral species may be detected on or in crevicular epithelial cells from sites with periodontitis and from periodontally healthy sulci.

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Section: Introductionmentioning

confidence: 96%

Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions

Colombo¹,

Silva²,

Haffajee

et al. 2006

Journal of Medical Microbiology

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“…Indeed, intracellular P. gingivalis cells remain viable and capable of spreading between host cells [5], and epithelial cells can survive for prolonged periods after infection with P. gingivalis [6]. Moreover, epithelial cells recovered from the oral cavity show high levels of intracellular P. gingivalis [7][8][9], indicating that an intracellular location is an integral component of the lifestyle of this organism and contributes to persistence in the oral cavity.…”

Section: Introductionmentioning

confidence: 99%

Quantitative proteomics of intracellular Porphyromonas gingivalis

Xia¹,

Wang²,

Taub³

et al. 2007

Proteomics

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Whole-cell quantitative proteomic analyses were conducted to investigate the change from an extracellular to intracellular lifestyle for Porphyromonas gingivalis, a Gram-negative intracellular pathogen associated with periodontal disease. Global protein abundance data for P. gingivalis strain ATCC 33277 internalized for 18 hours within human gingival epithelial cells and controls exposed to gingival cell culture medium were obtained at sufficient coverage to provide strong evidence that these changes are profound. A total of 385 proteins were over-expressed in internalized P. gingivalis relative to controls; 240 proteins were shown to be under-expressed. This represented in total about 28% of the protein encoding ORFs annotated for this organism, and slightly less than half of the proteins that were observed experimentally. Production of several proteases, including the classical virulence factors RgpA, RgpB, and Kgp, was decreased. A separate validation study was carried out in which a 16-fold dilution of the P. gingivalis proteome was compared to the undiluted sample in order to assess the quantitative false negative rate (all ratios truly alternative). Truly null (no change) abundance ratios from technical replicates were used to assess the rate of quantitative false positives over the entire proteome. A global comparison between the direction of abundance change observed and previously published bioinformatic gene pair predictions for P. gingivalis will assist with future studies of P. gingivalis gene regulation and operon prediction.

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“…The best hybridization conditions were found to be at a temperature of 59 • C for 150 min, which make our FISH protocol as fast as the previously reported for DNA-FISH (Zijnge et al, 2010;Rudney et al, 2001;Colombo et al, 2007;Stoltenberg et al, 1993;Sunde et al, 2003;Rudney et al, 2005a,b;Johnson et al, 2008) and suitable both for clinical, if it proves useful (Colombo et al, 2009;Sayehmiri et al, 2015;Hong et al, 2015), and research purposes.…”

Section: Discussionmentioning

confidence: 55%

“…The PgPNA1007 probe has a higher theoretical performance in P. gingivalis detection compared with the DNA probes used in several previous studies (Rudney et al, 2001;Colombo et al, 2007;Stoltenberg et al, 1993;Sunde et al, 2003;Rudney et al, 2005a,b;Johnson et al, 2008). Its theoretical performance is only comparable to the DNA probe previously developed by Sunde et al (2003).…”

Section: Discussionmentioning

confidence: 78%

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Novel strategy to detect and locate periodontal pathogens: The PNA-FISH technique

Mendes

Rocha

Azevedo

et al. 2016

Microbiological Research

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Purpose: We aim to develop peptic nucleic acid (PNA) probes for the identification and localization of Aggregatibacter actinomycetemcomintans and Porphyromonas gingivalis in sub-gingival plaque and gingival biopsies by Fluorescence in situ Hybridization (FISH).Methods: A PNA probe was designed for each microorganism. The PNA-FISH method was optimized to allow simultaneous hybridization of both microorganisms with their probe (PNA-FISH multiplex). After being tested on representative strains of P. gingivalis and A. actinomycetemcomitans, the PNA-FISH method was then adapted to detect microorganisms in the subgingival plaque and gingival samples, collected from patients with severe periodontitis. Results:The best hybridization conditions were found to be 59 • C for 150 min for both probes (PgPNA1007 and AaPNA235). The in silico sensitivity and specificity was both 100% for PgPNA1007 probe and 100% and 99.9% for AaPNA235 probe, respectively. Results on clinical samples showed that the PNA-FISH method was able to detect and discriminate target bacteria in the mixed microbial population of the subgingival plaque and within periodontal tissues.Conclusion: This investigation presents a new highly accurate method for P. gingivalis and A. actino-mycetemcomitans detection and co-location in clinical samples, in just few hours. With this technique we were able to observe spatial distribution of these species within polymicrobial communities in the periodontal pockets and, for the first time with the FISH method, in the organized gingival tissue.2

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Intracellular Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in Buccal Epithelial Cells Collected from Human Subjects

Cited by 158 publications

References 43 publications

Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions

Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions

Quantitative proteomics of intracellular Porphyromonas gingivalis

Novel strategy to detect and locate periodontal pathogens: The PNA-FISH technique

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