Identification of oral bacteria associated with crevicular epithelial cells from chronic periodontitis lesions.
TL;DR: It is 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.
Abstract: 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 > or =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.
Citations
More filters
TL;DR: Current knowledge and emerging mechanisms governing oral polymicrobial synergy and dysbiosis that have both enhanced the understanding of pathogenic mechanisms and aided the design of innovative therapeutic approaches for oral diseases are discussed.
Abstract: The dynamic and polymicrobial oral microbiome is a direct precursor of diseases such as dental caries and periodontitis, two of the most prevalent microbially induced disorders worldwide. Distinct microenvironments at oral barriers harbour unique microbial communities, which are regulated through sophisticated signalling systems and by host and environmental factors. The collective function of microbial communities is a major driver of homeostasis or dysbiosis and ultimately health or disease. Despite different aetiologies, periodontitis and caries are each driven by a feedforward loop between the microbiota and host factors (inflammation and dietary sugars, respectively) that favours the emergence and persistence of dysbiosis. In this Review, we discuss current knowledge and emerging mechanisms governing oral polymicrobial synergy and dysbiosis that have both enhanced our understanding of pathogenic mechanisms and aided the design of innovative therapeutic approaches for oral diseases.
938 citations
TL;DR: Evidence for microbial maternal transmission is increasingly widespread across animals, and collective knowledge compels a paradigm shift—one in which maternal transmission of microbes advances from a taxonomically specialized phenomenon to a universal one in animals.
Abstract: Summary The sterile womb paradigm is an enduring premise in biology that human infants are born sterile. Recent studies suggest that infants incorporate an initial microbiome before birth and receive copious supplementation of maternal microbes through birth and breastfeeding. Moreover, evidence for microbial maternal transmission is increasingly widespread across animals. This collective knowledge compels a paradigm shift—one in which maternal transmission of microbes advances from a taxonomically specialized phenomenon to a universal one in animals. It also engenders fresh views on the assembly of the microbiome, its role in animal evolution, and applications to human health and disease.
655 citations
Cites background from "Identification of oral bacteria ass..."
...[3], digestion [4,5], immune cell development [6–8], dental health [9,10], and resistance to pathogens [11,12]....
[...]
TL;DR: Differences in microbial composition and function are being linked to health and disease states in different organs and tissues in different body habitats.
Abstract: Mammals are complex assemblages of mammalian and bacterial cells organized into functional organs, tissues, and cellular communities. Human biology can no longer concern itself only with human cells: Microbiomes at different body sites and functional metagenomics must be considered part of systems biology. The emergence of metagenomics has resulted in the generation of vast data sets of microbial genes and pathways present in different body habitats. The profound differences between microbiomes in various body sites reveal how metagenomes contribute to tissue and organ function. As next-generation DNA-sequencing methods provide whole-metagenome data in addition to gene-expression profiling, metaproteomics, and metabonomics, differences in microbial composition and function are being linked to health and disease states in different organs and tissues. Global parameters of microbial communities may provide valuable information regarding human health status and disease predisposition. More detailed knowledge...
460 citations
TL;DR: Rheumatoid arthritis (RA) is now clearly a true autoimmune disease with accumulating evidence of pathogenic disease‐specific autoimmunity to citrullinated proteins, and both smoking and Porphyromonas gingivalis are attractive etiological agents for further investigation into the gene/environment/autoimmunities triad of RA.
Abstract: Rheumatoid arthritis (RA) is now clearly a true autoimmune disease with accumulating evidence of pathogenic disease-specific autoimmunity to citrullinated proteins. Citrullination, also termed deimination, is a modification of arginine side chains catalyzed by peptidylarginine deiminase (PAD) enzymes. This post-translational modification has the potential to alter the structure, antigenicity, and function of proteins. In RA, antibodies to cyclic citrullinated peptides are now well established for clinical diagnosis, though we argue that the identification of specific citrullinated antigens, as whole proteins, is necessary for exploring pathogenic mechanisms. Four citrullinated antigens, fibrinogen, vimentin, collagen type II, and alpha-enolase, are now well established, with others awaiting further characterization. All four proteins are expressed in the joint, and there is evidence that antibodies to citrullinated fibrinogen and collagen type II mediate inflammation by the formation of immune complexes, both in humans and animal models. Antibodies to citrullinated proteins are associated with HLA 'shared epitope' alleles, and autoimmunity to at least one antigenic sequence, the CEP-1 peptide from citrullinated alpha-enolase (KIHAcitEIFDScitGNPTVE), shows a specific association with HLA-DRB1*0401, *0404, 620W PTPN22, and smoking. Periodontitis, in which Porphyromonas gingivalis is a major pathogenic bacterium, has been linked to RA in epidemiological studies and also shares similar gene/environment associations. This is also the only bacterium identified that expresses endogenous citrullinated proteins and its own bacterial PAD enzyme, though the precise molecular mechanisms of bacterial citrullination have yet to be explored. Thus, both smoking and Porphyromonas gingivalis are attractive etiological agents for further investigation into the gene/environment/autoimmunity triad of RA.
444 citations
TL;DR: The oral cavity of healthy individuals contains hundreds of different bacterial, viral, and fungal species, many of which can associate to form biofilms, which are resistant to mechanical stress or antibiotic treatment.
Abstract: The oral cavity of healthy individuals contains hundreds of different bacterial, viral, and fungal species. Many of these can associate to form biofilms, which are resistant to mechanical stress or antibiotic treatment. Most are also commensal species, but they can become pathogenic in responses to changes in the environment or other triggers in the oral cavity, including the quality of an individual's personal hygiene. The complexity of the oral microbiome is being characterized through the newly developed tools of metagenomics. How the microbiome of the oral cavity contributes to health and disease is attracting the interest of a growing number of cell biologists, microbiologists, and immunologists.
397 citations
References
More filters
TL;DR: The purpose of the present investigation was to attempt to define communities using data from large numbers of plaque samples and different clustering and ordination techniques, which related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.
Abstract: It has been recognized for some time that bacterial species exist in complexes in subgingival plaque. The purpose of the present investigation was to attempt to define such communities using data from large numbers of plaque samples and different clustering and ordination techniques. Subgingival plaque samples were taken from the mesial aspect of each tooth in 185 subjects (mean age 51 +/- 16 years) with (n = 160) or without (n = 25) periodontitis. The presence and levels of 40 subgingival taxa were determined in 13,261 plaque samples using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments were made at 6 sites per tooth at each visit. Similarities between pairs of species were computed using phi coefficients and species clustered using an averaged unweighted linkage sort. Community ordination was performed using principal components analysis and correspondence analysis. 5 major complexes were consistently observed using any of the analytical methods. One complex consisted of the tightly related group: Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola. The 2nd complex consisted of a tightly related core group including members of the Fusobacterium nucleatum/periodonticum subspecies, Prevotella intermedia, Prevotella nigrescens and Peptostreptococcus micros. Species associated with this group included: Eubacterium nodatum, Campylobacter rectus, Campylobacter showae, Streptococcus constellatus and Campylobacter gracilis. The 3rd complex consisted of Streptococcus sanguis, S. oralis, S. mitis, S. gordonii and S. intermedius. The 4th complex was comprised of 3 Capnocytophaga species, Campylobacter concisus, Eikenella corrodens and Actinobacillus actinomycetemcomitans serotype a. The 5th complex consisted of Veillonella parvula and Actinomyces odontolyticus. A. actinomycetemcomitans serotype b, Selenomonas noxia and Actinomyces naeslundii genospecies 2 (A. viscosus) were outliers with little relation to each other and the 5 major complexes. The 1st complex related strikingly to clinical measures of periodontal disease particularly pocket depth and bleeding on probing.
4,143 citations
"Identification of oral bacteria ass..." refers background or methods or result in this paper
...Specific groups of micro-organisms in the subgingival biofilm described by Socransky et al. (1998) as members of the ‘red and orange complexes’ have been commonly associated with destructive periodontal disease....
[...]
...Periodontal diseases are characterized by chronic inflammatory lesions and destruction of the supporting periodontal tissues, which are associated with specific microbial complexes in subgingival biofilms (Socransky et al., 1998)....
[...]
...The checkerboard technique has been widely used to identify microbial complexes within dental biofilms, as well as those associated with different oral epithelia in various clinical conditions (Socransky et al., 1998; Colombo et al., 2002; Mager et al., 2003)....
[...]
...These results are consistent with other studies that associate a greater frequency of streptococci with periodontal health (Haffajee et al., 1997; Dibart et al., 1998; Socransky et al., 1998)....
[...]
713 citations
"Identification of oral bacteria ass..." refers background in this paper
...The full mouth periodontal destruction patterns, including mean PPD (3?4± 0?2 mm), mean CAL (3?6±0?2 mm), mean percentage of sites with BOP (40±4?1), SB (44±4?3) and SUP (1?3±0?2) and mean number of missing teeth (4?7±2), indicated that most of the subjects presented a moderate to severe form of chronic periodontitis (Armitage, 2004)....
[...]
...7±2), indicated that most of the subjects presented a moderate to severe form of chronic periodontitis (Armitage, 2004)....
[...]
Journal Article•
TL;DR: The method permits the simultaneous determination of the presence of multiple bacterial species in single or multiple dental plaque samples, thus suggesting its usefulness for a range of clinical or environmental samples.
Abstract: A method is introduced for hybridizing large numbers of DNA samples against large numbers of DNA probes on a single support membrane. Denatured DNA from up to 43 samples was fixed in separate lanes on a single membrane mounted in a Miniblotter 45. The membrane was then rotated 90 degrees in the same device, which enabled simultaneous hybridization with 43 different DNA probes. Hybridizations were also performed on lysates of bacterial cells blotted to membranes. A MiniSlot device allowed lysates loaded in parallel channels to be aspirated through the membrane, depositing horizontal lanes on the membrane surface. Hybridizations were performed in vertical lanes with either digoxigenin-labeled whole genomic probes or 16S rRNA-based oligonucleotide probes directly conjugated to alkaline phosphatase. The method permits the simultaneous determination of the presence of multiple bacterial species in single or multiple dental plaque samples, thus suggesting its usefulness for a range of clinical or environmental samples.
677 citations
"Identification of oral bacteria ass..." refers methods in this paper
...…Rockville, MD. DThe Forsyth Institute, Boston, MA. 610 Journal of Medical Microbiology 55 Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Wed, 05 Dec 2018 22:27:13 (Haffajee et al., 1997) of the checkerboard DNA–DNA hybridization method described by Socransky et al. (1994)....
[...]
TL;DR: Invasion was an active process requiring both bacterial and epithelial cell energy production and was inhibited by protease inhibitors, suggesting that P. gingivalis proteases may be involved in the invasion process.
Abstract: Porphyromonas gingivalis, a periodontal pathogen, can invade primary cultures of gingival epithelial cells. Optimal invasion occurred at a relatively low multiplicity of infection (i.e., 100) and demonstrated saturation at a higher multiplicity of infection. Following the lag phase, during which bacteria invaded poorly, invasion was independent of growth phase. P. gingivalis was capable of replicating within the epithelial cells. Invasion was an active process requiring both bacterial and epithelial cell energy production. Invasion was sensitive to inhibitors of microfilaments and microtubules, demonstrating that epithelial cell cytoskeletal rearrangements are involved in bacterial entry. P. gingivalis, but not epithelial cell, protein synthesis was necessary for invasion. Invasion within the epithelial cells was not blocked by inhibitors of protein kinase activity. Invasion was inhibited by protease inhibitors, suggesting that P. gingivalis proteases may be involved in the invasion process. Low-passage clinical isolates of P. gingivalis invaded with higher efficiency than the type strain. Serum inhibited invasion of the type strain but had no effect on the invasion of a clinical isolate. Invasion of gingival epithelial cells by P. gingivalis may contribute to the pathology of periodontal diseases.
516 citations
"Identification of oral bacteria ass..." refers background in this paper
...nucleatum, are capable of elaborating a vast array of virulence factors that allow them to adhere to and invade the host cells, avoid or resist the immune response, and cause periodontal tissue damage (Lamont et al., 1995; Madianos et al., 1996; Han et al., 2000; Rudney et al., 2001, 2005; Vitkov et al., 2005)....
[...]
..., 2001), Porphyromonas gingivalis (Sandros et al., 1994; Lamont et al., 1995; Madianos et al., 1996), Fusobacterium nucleatum (Han et al....
[...]
...…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.,…...
[...]
...…capable of elaborating a vast array of virulence factors that allow them to adhere to and invade the host cells, avoid or resist the immune response, and cause periodontal tissue damage (Lamont et al., 1995; Madianos et al., 1996; Han et al., 2000; Rudney et al., 2001, 2005; Vitkov et al., 2005)....
[...]
TL;DR: Clinical improvement post-SRP was accompanied by a modest change in theSubgingival microbiota, primarily a reduction in P. gingivalis, B. forsythus and T. denticola, suggesting potential targets for therapy and indicating that radical alterations in the subgingival bacteria may not be necessary or desirable in many patients.
Abstract: The purpose of the present investigation was to examine the effect of SRP on clinical and microbiological parameters in 57 subjects with adult periodontitis (mean age 47 +/- 11 years). Subjects were monitored clinically and microbiologically prior to and 3, 6 and 9 months after full-mouth SRP under local anaesthesia. Clinical assessments of plaque, redness, suppuration, BOP, pocket depth and attachment level were made at 6 sites per tooth. The means of duplicate attachment level measurements taken at each visit were used to assess change between visits. Clinical data were averaged within each subject and then averaged across subjects for each visit. Subgingival plaque samples were taken from the mesial aspect of each tooth and the presence and levels of 40 subgingival taxa were determined using whole genomic DNA probes and checkerboard DNA-DNA hybridization. The mean levels and % of sites colonized by each species (prevalence) was computed for each subject at each visit. Differences in clinical and microbiological parameters before and after SRP were sought using the Wilcoxon signed ranks test or the Quade test for more than 2 visits. Overall, there was a mean gain in attachment level of 0.11 +/- 0.23 mm (range -0.53 to 0.64 mm) 3 months post-therapy. There was a significant decrease in the % of sites exhibiting gingival redness (68 to 57%) and BOP (58 to 52%) as well as a mean (+/-SEM) pocket depth (3.3 +/- 0.06 to 3.1 +/- 0.05 mm). Sites with pre-therapy pocket depths of 6 mm pockets showed a significant decrease in pocket depth and attachment level measurements post-therapy. Significant clinical improvements were seen in subjects who had never smoked or were past smokers but not in current smokers. Mean prevalences and levels of P. gingivalis, T. denticola and B. forsythus were significantly reduced after SRP, while A. viscosus showed a significant increase in mean levels. The mean decrease in prevalence of P. gingivalis was similar at all pocket depth categories, while B. forsythus decreased more at shallow and intermediate pockets and A. viscosus increased most at deep sites. P. gingivalis. B. forsythus and T. denticola were equally prevalent among current, past and never smokers pre-therapy, decreased significantly post-SRP in never and past smokers but increased in current smokers. Clinical improvement post-SRP was accompanied by a modest change in the subgingival microbiota, primarily a reduction in P. gingivalis, B. forsythus and T. denticola, suggesting potential targets for therapy and indicating that radical alterations in the subgingival microbiota may not be necessary or desirable in many patients.
456 citations
"Identification of oral bacteria ass..." refers background or methods or result in this paper
...11 On: Wed, 05 Dec 2018 22:27:13 (Haffajee et al., 1997) of the checkerboard DNA–DNA hybridization method described by Socransky et al....
[...]
...Microbial data were expressed as the mean percentage of colonized sites (prevalence) and mean levels (6104 cells) of each species in the 49 subjects (Haffajee et al., 1997; Colombo et al., 2002)....
[...]
...Microbial data were expressed as the mean percentage of colonized sites (prevalence) and mean levels (610(4) cells) of each species in the 49 subjects (Haffajee et al., 1997; Colombo et al., 2002)....
[...]
...…Rockville, MD. DThe Forsyth Institute, Boston, MA. 610 Journal of Medical Microbiology 55 Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Wed, 05 Dec 2018 22:27:13 (Haffajee et al., 1997) of the checkerboard DNA–DNA hybridization method described by Socransky et al. (1994)....
[...]
...These results are consistent with other studies that associate a greater frequency of streptococci with periodontal health (Haffajee et al., 1997; Dibart et al., 1998; Socransky et al., 1998)....
[...]