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Architecture of Intact Natural Human Plaque Biofilms Studied by Confocal Laser Scanning Microscopy
S.R. Wood
Division of Oral Biology
J. Kirkham
Division of Oral Biology
P.D. Marsh
Division of Oral Biology
R.C. Shore
Division of Oral Biology
B. Nattress
Division of Restorative Dentistry, Leeds Dental Institute, University of Leeds, Clarendon Way, Leeds LS2 9LU, United Kingdom
C. Robinson
Division of Oral Biology
Determination of the structure of human plaque will be of great benefit in the prediction of its formation and also the effects of treatment. However, a problem lies in the harvesting of undisturbed intact plaque samples from human volunteers and the viewing of the biofilms in their natural state. In this study, we used an in situ device for the in vivo generation of intact dental plaque biofilms on natural tooth surfaces in human subjects. Two devices were placed in the mouths of each of eight healthy volunteers and left to generate biofilm for 4 days. Immediately upon removal from the mouth, the intact, undisturbed biofilms were imaged by the non-invasive technique of confocal microscopy in both reflected light and fluorescence mode. Depth measurements indicated that the plaque formed in the devices was thicker round the edges at the enamel/nylon junction (range = 75-220 µm) than in the center of the devices (range = 35-215 µm). The reflected-light confocal images showed a heterogeneous structure in all of the plaque biofilms examined; channels and voids were clearly visible. This is in contrast to images generated previously by electron microscopy, suggesting a more compact structure. Staining of the biofilms with fluorescein in conjunction with fluorescence imaging suggested that the voids were fluid-filled. This more open architecture is consistent with recent models of biofilm structure from other habitats and has important implications for the delivery of therapeutics to desired targets within the plaque.
Key Words: biofilm • plaque • architecture • confocal microscopy.
Journal of Dental Research, Vol. 79, No. 1,
21-27 (2000)
DOI: 10.1177/00220345000790010201
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