[Effects of NGF on proliferation, mitotic cycle, collagen synthesis and migration of human dermal fibroblasts in vitro].
OBJECTIVE
To investigate the effects of NGF on the proliferation, mitotic cycle, collagen synthesis and migration of human dermal fibroblasts (HDFs), and to explore the function of NGF on the wound healing.


METHODS
The 3rd generation of HDFs were incubated with various concentrations of NGF (0, 25, 50, 100, 200 and 400 ng/mL), the cell proliferation was measured with MTT assay. After treated with NGF at 0, 100 ng/mL, the cell cycle of HDFs was determined by flow cytometry (FCM). Hydroxyproline and real-time fluorescence quantitative PCR (FQ-PCR) were used to measure collagen synthesis at protein level and mRNA level respectively. The in vitro cell scratch wound model was set up to observe the effect of NGF (0, 50, 100 and 200 ng/mL) on the migration of HDFs after 24 hours of culture.


RESULTS
Absorbance value of HDFs for different concentrations of NGF (0, 25, 50, 100, 200, and 400 ng/mL) showed that NGF did not influence the proliferation of HDFs (P > 0.05). When HDFs were treated with NGF at 0 and 100 ng/mL, the result of FCM analysis showed that percentage of HDFs in G0/G1, S, G2/M phases were not changed (P > 0.05). Compared with control group, the expression of Col I and Col III were not significantly different, measured by both hydroxyproline and FQ-PCR (P > 0.05). The rates of HDFs' migration at various concentrations of NGF (0, 50, 100, 200 ng/mL) were 52.12% +/- 6.50%, 80.67% +/- 8.51%, 66.33% +/- 3.58%, and 61.19% +/- 0.97%, respectively, indicating that NGF could significantly enhanced the migration of HDFs at 50 and 100 ng/mL (P < 0.05).


CONCLUSIONS
NGF does not influence proliferation, mitotic cycle and collagen synthesis of HDFs, but significantly enhanced migration in an in vitro model of wounded fibroblasts.
Gan
Huali
H
Division of Stem Cells and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu Sichuan 610041, PR China.
Xie
Huiqi
H
Chen
Xiaohe
X
Luo
Jingcong
J
D002453:Cell Cycle / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020932:Nerve Growth Factor / Q000494:pharmacology*
D003094:Collagen; D020932:Nerve Growth Factor