Methods: Periodontal ligament-endothelial cells (PDL-EC) and gingiva-endothelial cells (G-EC) were isolated by coupling to monoclonal anti-CD31 magnetic beads. Both PDL-EC and G-EC were characterized to definitively demonstrate the culture represented pure EC. All EC were seeded on 96-well plate coated with collagen I, collagen IV, fibronectin, gelatin, and poly-D-lysine (PDL) as well as uncoated 96-well plate. Then, they were cultured in EGM®-2MV media for 2, 6, 24 hours (cell attachment), and, for 1, 2, 3, 5, 7 days (cell proliferation). Their growth was determined by resazurin reduction assay (AlamarBlue®) and direct cell counting. Data were shown as growth curves and the percentage of cell attachment and cell proliferation, respectively.
Results: Both PDL-EC and G-EC revealed the specific EC characteristics and showed more slowly growth rate than HUVEC in all ECM protein coating. For cell attachment (2 and 6h), only collagen I increased significantly (p<0.05) the % cell attachment of PDL-EC and G-EC while gelatin and PDL decreased significantly (p<0.05) the % cell attachment of those, as comparing to uncoated. For proliferation, in addition to collagen I, all ECM protein decreased significantly (p<0.05) the % cell proliferation of PDL-EC and G-EC, even though sometimes no statistic difference was detected.
Conclusions: ECM protein influence on growth of cultured periodontal-derived EC in vitro. Collagen I is an optimal substratum to enhance those cell growth.