Effects of serum on hepatocyte growth factor secretion and activation by periodontal ligament and gingival fibroblasts.
BACKGROUND
Hepatocyte growth factor (HGF)/scatter factor is a paracrine growth factor secreted by mesenchymal cells, which exerts an effect on a variety of epithelial cell types. Our recent study revealed that periodontal ligament fibroblasts (PLF) and gingival fibroblasts (GF) cultured in the presence of serum which contains various stimulants produced HGF or HGF-like factor, a predominant chemoattractant for gingival epithelial cells, and suggested that it could be involved in epithelial down-growth in periodontitis.


METHODS
To clarify whether serum in medium stimulates PLF and GF to synthesize or activate HGF, the effect of fetal bovine serum (FBS) on HGF production was determined by enzyme-linked immunoabsorbent assay (ELISA), and its chemotactic activity for gingival epithelial cells was examined by modified Boyden chamber assay.


RESULTS
One to 10% FBS in the culture medium stimulated HGF secretion in a dose-dependent manner and the chemotactic activity was decreased by treatment with anti-hHGF neutralizing antibody. Furthermore, fibroblast-conditioned medium incubated with FBS and aprotinin reduced its chemotactic activity. Interestingly, serum-free culture of PLF and GF produced potent chemoattractants for gingival epithelial cells other than HGF.


CONCLUSIONS
These results show that FBS stimulates both HGF secretion and activation by PLF and GF.
Ohshima
Mitsuhiro
M
Department of Biochemistry, Nihon University School of Dentistry, Tokyo, Japan.
Yokosuka
Ryoji
R
Yamazaki
Yoshitaka
Y
Tokunaga
Tsuyoshi
T
Maeno
Masao
M
Otsuka
Kichibee
K
D000818:Animals; D000906:Antibodies; D007611:Aprotinin / Q000494:pharmacology; D001769:Blood; D002417:Cattle; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology; D002633:Chemotaxis / Q000187:drug effects; D018920:Coculture Techniques; D003470:Culture Media; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D015194:Diffusion Chambers, Culture; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D017228:Hepatocyte Growth Factor / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D019899:Paracrine Communication / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology / Q000378:metabolism*; D013223:Statistics as Topic
D000906:Antibodies; D002630:Chemotactic Factors; D003470:Culture Media; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D017228:Hepatocyte Growth Factor; D007611:Aprotinin