[Effects of basic fibroblast growth factor and vascular endothelial growth factor on the proliferation, migration and adhesion of human periodontal ligament stem cells in vitro].

Objective To evaluate the effects of basic fibroblast growth factor(FGF-2) and vascular endothelial growth factor (VEGF) on the proliferation,migration,and adhesion of human periodontal ligament stem cells(PDLSC) in vitro.Methods Human PDLSC were cultured in vitro using tissue culture method.The cells were cultured and incubated with various concentrations of FGF-2 and VEGF [A:α-MEM with 2％ fetal bovine serum(FBS) (control 1) ; B:A supplemented with 20 μg/L FGF-2 ; C:A supplemented with 10 μg/L VEGF;D:A supplemented with 20 μg/L FGF-2 and 10 μg/L VEGF; E:α-MEM with 10％ FBS(control 2) ; F:E supplemented with 20 μg/L FGF-2 ; G:E supplemented with 10 μg/L VEGF; H:E supplemented with 20 μg/L FGF-2 and 10 μg/L VEGF].Soluble tetrazolium salts assay was used to evaluate the proliferative capacity on the 1st,3rd,5th and 7th d.Then the groups were changed according to result of the proliferation assay (control:α-MEM with 2％ FBS; FGF-2 group:control supplemented with 20 μg/L FGF-2 ; VEGF:control supplemented with 10 μg/L VEGF ; Combination group:control supplemented with 20 μg/L FGF-2 and 10 μg/L VEGF).The cell cycle,migration and adhesion capacities were evaluated using flow cytometer,soluble tetrazolium salts assay,cell adhesion assay and scratch woundhealing motility assay.Results In 2％ volume fraction serum containing medium,FGF-2 and VEGF did not stimulate the cell proliferation.However,in 10％ serum condition,in groups treated with FGF-2 for 3,5 or 7 d,the A value was (1.22 ± 0.17,2.15 ± 0.19,2.72 ± 0.11) respectively,which were significantly higher than that in the control group (0.76 ± 0.16,1.25 ± 0.06,1.64 ±.0.09) (P ＜ 0.01) while lower than that in the group treated with FGF-2 and VEGF in combination on the 5 th and 7 th d (2.46 ±0.17,3.18 ±0.27) (P ＜0.05).The A value in the VEGF group on the 5 th and 7 th d is higher than the control group while lower than the FGF-2 group(1.66 ± 0.05,2.13 ± 0.13) (P ＜ 0.05).Flow cytometer showed that the proliferation index in VEGF group [(34.3 ±2.0)％] were significantly lower than those in FGF-2 [(46.8 ±3.2)％] group and (FGF-2 + VEGF) group[(45.0 ±4.0)％] but higher than in the control group [(14.5 ± 1.7)％] (P ＜ 0.01).The cell migration assay indicated that the group stimulated with FGF-2 showed no migration promoted effect.Cell adhesion assay showed that the ratio of the adhesive cells number to the original cells number is greater in the FGF-2 group(79 ±4) than in the VEGF group(62 ±4) (P ＜ 0.05).Light microscope identified a better cellular morphology on the adhesive surface in the group with FGF-2 than groups without FGF-2.Conclusions Both FGF-2 and VEGF could simulate the proliferation of PDLSC in a dose dependent manner,and showed an synergistic effect.FGF-2 was more effective to promote the adhesive capacity of PDLSC compared with VEGF.VEGF could facilitate the migration of PDLSC to the wound side. Key words: Fibroblast growth factor 2;  Vascular endothelial growth factors;  Periodontal regeneration;  Periodontal ligament stem cells