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Porphyromonas gingivalis (P. gingivalis) has been implicated as an important pathogen in periodontal diseases, and its protease (Arg-gingipain) is one of the most important disease-causing factors. We have examined the effect of Arg-gingipain on the cell adhesion molecules of human periodontal ligament fibroblasts (HPLF). The Arg-gingipain was purified from the culture supernatant of P. gingivalis. Confluent monolayer cultures of HPLF were incubated for up to 24 hr with Arg-gingipain. As for the morphological changes, HPLF detached from the dish at 16 hr. The growth rate as evaluated by the MTT assay increased at 12 hr, and then the rate decreased. The cell adhesion molecules (ICAM-1, intercellular adhesion molecule-1; CD54, VCAM-1, vascular cell adhesion molecule-1; CD106, and VLA-4, very late antigen-4; CD49d/cd29) were constitutively expressed on HPLF by flow cytometric analysis. The amounts of fibronectin were increased at 12 hr and then, its production was decreased. The levels of ICAM-1, VCAM-1 and VLA-4 expressions were increased; however, after 16 hr, the levels of expression of all cell adhesion molecules were decreased. Our experiments showed that adhesion decreases as a result of destruction of the extracellular matrix when bacterial protease brings about rapid cell destruction. This may suggest that the increase of expression of cell adhesion molecules is due to a mechanism that compensates for the decreased cellular-extracellular matrix. Alternatively, it may suggest that after the expression is activated, cell adhesion molecules help to increase bacterial adhesion to cells.

Effect of proteases from Porphyromonas gingivalis on adhesion molecules of human periodontal ligament fibroblast cells.