Approximately 1/3 of the population is affected by chronic periodontitis, a bacterially induced destruction of the attachment of the tooth to bone. The gingival crevice is host to a large number of bacteria, most of them currently uncultivated. The pathogenesis of periodontitis, although not well understood, is thought to involve polymicrobial consortia that appear to include many of these uncultivated bacteria. The gingival crevice offers colonizing bacteria a complex environment that includes a non-shedding tooth surface, a continuously desquamating epithelial wall covering the underlying connective tissue, and the narrow fluid-filled space in between. Many species of bacteria have been shown to attach to or invade the tissues, and these invasive bacteria are probably shed from the tooth-associated biofilm. Both the biofilm community and invasive bacteria are assumed to play an important role in pathogenesis. The goal of this project is to compare the bacterial community profiles found in periodontal health and disease from each of the physical compartments in the gingival crevice: the tooth, the crevicular fluid, and the tissue. Comparisons across disease status and physical location will elucidate the most important species and the most critical physical location for pathogens or beneficial species and further our understanding of the dynamics within this complex microbial community. A combination of rDNA 16S cloning and sequencing, PCR, and real-time PCR will be used for these studies. Experiments to further our understanding of polymicrobial interactions are also planned. Data on presence of cultivated and uncultivated species will be obtained by PCR, and analyzed for bacterial co- associations using a new statistical approach, Cluster Analysis for Categorical Data (CACD). This approach was developed by our research team to determine physically co-associated complexes of bacteria. These experiments should bring us closer to understanding the complex interactions that make up the crevicular ecosystem and biofilm. Finally, using the tools developed in the first two aims we will cultivate and characterize as-yet-uncultivated species that show associations with periodontitis or health. We will isolate these bacterial species using a combination of newly developed cultivation procedures and molecular tracking methods. The proposed studies will bring us closer to understanding the bacterial etiology of periodontitis, and closer to targeted therapies to promote health and treat disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE010467-14
Application #
8082736
Study Section
Oral, Dental and Craniofacial Sciences Study Section (ODCS)
Program Officer
Lunsford, Dwayne
Project Start
1995-02-01
Project End
2013-05-31
Budget Start
2011-06-01
Budget End
2013-05-31
Support Year
14
Fiscal Year
2011
Total Cost
$345,466
Indirect Cost
Name
Ohio State University
Department
Dentistry
Type
Schools of Dentistry
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Griffen, Ann L; Beall, Clifford J; Campbell, James H et al. (2012) Distinct and complex bacterial profiles in human periodontitis and health revealed by 16S pyrosequencing. ISME J 6:1176-85
Griffen, Ann L; Beall, Clifford J; Firestone, Noah D et al. (2011) CORE: a phylogenetically-curated 16S rDNA database of the core oral microbiome. PLoS One 6:e19051
Igboin, Christina O; Moeschberger, Melvin L; Griffen, Ann L et al. (2011) Porphyromonas gingivalis virulence in a Drosophila melanogaster model. Infect Immun 79:439-48
Igboin, Christina O; Tordoff, Kevin P; Moeschberger, Melvin L et al. (2011) Porphyromonas gingivalis-host interactions in a Drosophila melanogaster model. Infect Immun 79:449-58
Igboin, Christina O; Griffen, Ann L; Leys, Eugene J (2009) Porphyromonas gingivalis strain diversity. J Clin Microbiol 47:3073-81
Kumar, Purnima S; Griffen, Ann L; Moeschberger, Melvin L et al. (2005) Identification of candidate periodontal pathogens and beneficial species by quantitative 16S clonal analysis. J Clin Microbiol 43:3944-55
Kumar, P S; Griffen, A L; Barton, J A et al. (2003) New bacterial species associated with chronic periodontitis. J Dent Res 82:338-44
Leys, Eugene J; Lyons, Sharon R; Moeschberger, Melvin L et al. (2002) Association of Bacteroides forsythus and a novel Bacteroides phylotype with periodontitis. J Clin Microbiol 40:821-5
Becker, Mitzi R; Paster, Bruce J; Leys, Eugene J et al. (2002) Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol 40:1001-9
Rumpf, R W; Griffen, A L; Leys, E J (2000) Phylogeny of Porphyromonas gingivalis by ribosomal intergenic spacer region analysis. J Clin Microbiol 38:1807-10

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