As an essential first step in understanding the interaction between subgingival species and gingival epithelial cells, is to determine which species are present on or in epithelial cells recovered from the human gingival crevice or periodontal pocket.
The Specific Aims of this project will be as follows: 1) to isolate and identify cultivable bacterial species which are attached to or invade into epithelial cells derived from the gingival crevice or periodontal pocket, 2) to seek uncultivable species which invade into or adhere to epithelial cells, 3) to enumerate predominant bacterial species, using DNA probes, which are on or in epithelial cells derived from subjects of different age and racial/ethnic groups and different states of health or disease and 4) to use light and electron microscopy to determine the species, numbers and location (on and/or in) of bacteria associated with crevicular epithelial cells. Epithelial cells will be isolated from samples taken from the gingival crevice or periodontal pocket of subjects of different ages, gender and racial/ethnic group. Initial cultural microbiology studies will be performed on samples from 32 subjects. Cultivable bacteria recovered from such samples will be used to define a set of DNA probes for subsequent Specific Aims. PCR techniques will be used to seek uncultivable species from harvested crevicular epithelial cells and DNA probes to such species added to the battery defined by cultural studies. The complete set of probes will be used to examine samples from a larger group of 60 Black, 60 Hispanic, 60 Asiatic and 60 White subjects. Each racial/ethnic group will consist of equal numbers of male and female, young and old and periodontally healthy and periodontally diseased subjects. 4 to 8 epithelial call samples will be taken from each subject and assayed for their content of up 50 species using the DNA probes. Finally, epithelial cell samples will be examined by light and electron microscopy for the number and location of bacteria of different morphotypes. Bacterial cells will be identified in such samples by immunocytochemical and/or in situ hybridization techniques.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
1R37DE010977-01A1
Application #
2131988
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1995-04-01
Project End
2000-03-31
Budget Start
1995-04-01
Budget End
1996-03-31
Support Year
1
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Forsyth Institute
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02142
Bogren, Anna; Teles, Ricardo; Torresyap, Gay et al. (2007) A three-year prospective study of adult subjects with gingivitis. I: clinical periodontal parameters. J Clin Periodontol 34:1-6
Teles, R P; Bogren, A; Patel, M et al. (2007) A three-year prospective study of adult subjects with gingivitis II: microbiological parameters. J Clin Periodontol 34:7-17
Haffajee, Anne D; Teles, Ricardo P; Socransky, Sigmund S (2006) The effect of periodontal therapy on the composition of the subgingival microbiota. Periodontol 2000 42:219-58
Teles, Ricardo P; Haffajee, Anne D; Socransky, Sigmund S (2006) Microbiological goals of periodontal therapy. Periodontol 2000 42:180-218
Socransky, Sigmund S; Haffajee, Anne D (2005) Periodontal microbial ecology. Periodontol 2000 38:135-87
Haffajee, A D; Japlit, M; Bogren, A et al. (2005) Differences in the subgingival microbiota of Swedish and USA subjects who were periodontally healthy or exhibited minimal periodontal disease. J Clin Periodontol 32:33-9
Gapski, R; Barr, J L; Sarment, D P et al. (2004) Effect of systemic matrix metalloproteinase inhibition on periodontal wound repair: a proof of concept trial. J Periodontol 75:441-52
Socransky, S S; Haffajee, A D; Smith, C et al. (2004) Use of checkerboard DNA-DNA hybridization to study complex microbial ecosystems. Oral Microbiol Immunol 19:352-62
Haffajee, A D; Bogren, A; Hasturk, H et al. (2004) Subgingival microbiota of chronic periodontitis subjects from different geographic locations. J Clin Periodontol 31:996-1002
Haffajee, A D; Uzel, N G; Arguello, E I et al. (2004) Clinical and microbiological changes associated with the use of combined antimicrobial therapies to treat ""refractory"" periodontitis. J Clin Periodontol 31:869-77

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