Figure 2.

Figure 2.

Increased endothelium permeability in vitro following treatment with Porphyromonas gingivalis whole cells and outer membrane vesicles (OMVs) is gingipain dependent. (A) Movement of fluorescently labeled 70 kDa dextran from the upper well to the lower well in a Transwell assay increased in a time-dependent manner in the absence of human microvascular endothelial cells (HMEC-1; insert only), whereas this movement was almost abolished when a confluent endothelium was cultured on the insert surface (monolayer). Endothelial monolayers were treated with (B) whole bacteria or (C) OMVs from either W83 or ΔK/R-ab for 1.5 h, and then dextran permeability across the endothelium was measured for up to 5 h; phosphate-buffered saline (PBS)–treated endothelium was used as controls. Increased endothelial permeability was significantly increased in a time-dependent manner following exposure to W83 when compared to ΔK/R-ab equivalents and untreated controls for both whole bacteria and OMVs. No significant differences were observed between ΔK/R-ab-treated and uninfected controls. Data are presented as mean ± SD of 3 independent experiments and were analyzed by 1-way analysis of variance followed by Tukey’s post hoc multiple comparisons test. *P < 0.05. **P < 0.01.