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RESEARCH REPORT |
1 Department of Oral Science, School of Dentistry, and
2 Biomedical Imaging and Processing Laboratory, Department of Neuroscience, University of Minnesota, 17-252 Moos Tower, 515 Delaware St. SE, Minneapolis, MN 55455, USA;
* corresponding author, jrudney{at}tc.umn.edu
Previously, we used in situ hybridization and confocal microscopy to detect the periodontal pathogens Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis within buccal epithelial cells taken directly from the mouth. This study tested the hypothesis that the intracellular flora of buccal cells is polymicrobial. Mixtures containing a red fluorescent universal probe paired with green fluorescent versions of either A. actinomycetemcomitans-, P. gingivalis-, or T. forsythensis-specific probes were hybridized with buccal cells collected from each of 38 healthy humans. We verified co-localization of probe pairs within cells by generating three-dimensional reconstructions. Intracellular bacteria were detected in every subject. Each cell that was labeled with a species-specific probe also contained bacteria recognized only by the universal probe. Bacteria labeled with specific probes often occupied smaller regions within larger masses of bacteria. Those findings suggest that future studies of invasion by oral bacteria may need to include microbial consortia.
KEY WORDS: polymicrobial • bacterial invasion • Actinobacillus actinomycetemcomitans • Porphyromonas gingivalis • Tannerella forsythensis
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