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 wound-healing 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.