Open AccessFluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells
Author(s) -
Belton Carol M.,
Izutsu Kenneth T.,
Goodwin Paul C.,
Park Yoonsuk,
Lamont Richard J.
Publication year - 1999
Publication title -
cellular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.542
H-Index - 138
eISSN - 1462-5822
pISSN - 1462-5814
DOI - 10.1046/j.1462-5822.1999.00022.x
Subject(s) - porphyromonas gingivalis , intracellular , biology , epithelium , fluorescence microscope , microbiology and biotechnology , bacteria , population , actin , in vitro , fluorescence , biochemistry , medicine , genetics , physics , environmental health , quantum mechanics
We have developed a fluorescence imaging technique using a DNA‐binding dye to visualize, over time, the physical interactions between Porphyromonas gingivalis and human gingival epithelial cells in vitro . The results extend previous observations of P. gingivalis invasion of gingival epithelial cells based on indirect measurements. An intracellular location for P. gingivalis was established by optical sectioning of images in the z ‐plane. Kinetic analysis showed that P. gingivalis invasion of epithelial cells is a rapid and efficient process, reaching completion after 12 min. Imaging of infected monolayers revealed that over 90% of a population of gingival epithelial cells contained bacteria. Furthermore, only vital bacteria were capable of invasion, and intracellular bacteria congregated in the perinuclear region of the epithelial cells. P. gingivalis remained inside the epithelial cells over a 24 h period and induced rearrangement of the actin cytoskeleton along with alteration of the size and shape of the epithelial cells. These findings provide direct evidence that entry rates of P. gingivalis into gingival epithelial cells are high and rapid, and that internalized bacteria initially localize in a specific region of the epithelial cells.