Assessing Internalized Porphyromonas gingivalis in Host Endothelial Cells
Transkript
During Porphyromonas gingivalis infection, the fimbriae on the anaerobic bacteria bind to specific receptors on host cells.
This binding initiates intracellular signaling cascades, resulting in actin rearrangement and bacterial internalization within phagosomes.
To assess the survival of internalized Porphyromonas gingivalis within endothelial cells in vitro, take a multi-well plate containing endothelial cells adhered to coverslips.
Add Porphyromonas gingivalis suspension. Incubate to facilitate bacterial internalization.
Remove media and wash using an anaerobic buffer to remove non-internalized bacteria.
Add media containing antibiotics, impermeable to the endothelial cells, to eliminate the surface-attached bacteria.
Incubate with a mild surfactant to lyse the endothelial cells, releasing the bacteria without causing their lysis.
Dilute the collected lysate with media. Prepare serial dilutions of the lysate.
Plate the dilutions on blood agar plates. Incubate under anaerobic conditions for Porphyromonas gingivalis to grow and form visible colonies.
Count the colonies to assess the ability of Porphyromonas gingivalis to invade and survive within endothelial cells.
