Characterization of periodontal cells in an in vitro healing model

Objectives: The differences between proliferation and migration of periodontal cells decided the result of periodontal regenerative therapy. An in vitro wound model was developed in our lab to show difference between gingival fibroblasts (GF) and periodontal ligament cells(PDLC) during the wound healing process. The aim of this research was to compare the levels of both proliferation and cellular wound filling between GF and PDL using our in vitro wound healing model.Methods: Primary culture of human PDLC and GF cells were established from explanted tissue, then they were passaged in 35mm culture dish with a cover glass(20x20)mm. Diploid cultures of both cell types were grown to confluence on the cover glasses, then the in vitro wounds were mechanically created by removing a 5mm wide band of the cell layer across the cover glasses on the dishes. The dishes were then incubated for 2,6,or 9 days in media containing 10% fetal bovine serum (FBS). At each time point, cells were fixed and stained with crystal blue. Cells were quantified within the boundaries by computer-assisted histomorphomertry.Results: The in vitro model for periodontal wound healing established in this research showed that GF filled in the wound site at a significantly faster than PDL at day 6, 9 .Conclusions: Our findings demonstrate the difference between PDL and GF with respect to proliferation and wound filling in an in vitro model, which suggests the regenerative process of PDLC maybe interfered by faster growth of GF.