Assessing Internalized Porphyromonas gingivalis in Host Endothelial Cells

Following infection of HUVEC cells, wash the cells three times with anaerobic PBS. Next, add 2 milliliters of VEGF medium supplemented with pre-optimized concentrations of antibiotics to the HUVEC cells. Incubate the cells for one hour. Aspirate the medium, and add 2 milliliters of 1% saponin solution in BHI to each well. Incubate for 15 minutes.

Next, scrape the lysed cells, and collect the lysate in a centrifuge tube. Dilute the lysate 1 to 1 with BHI medium, and then prepare serial dilutions starting at 1 to 100. Plate 200 microliters of the suitable concentrations on blood agar plates. Allow the bacteria to grow for seven days in an anaerobic chamber, and then count the colonies.