[Effects of fibroblast growth factor-10 on the secretions of transforming growth factor-alpha, platelet-derived growth factor-AB and vascular endothelial growth factor by normal adult human keratinocytes in culture].
OBJECTIVE
To evaluate the effect of fibroblast growth factor-10 (FGF-10) on the secretion of transforming growth factor-alpha (TGF-alpha), platelet-derived growth factor-AB (PDGF-AB) and vascular endothelial growth factor (VEGF) by human keratinocytes.


METHODS
Concentrations of FGF-10 used were 4, 16, 125 and 500 ng/ml. Serum-free keratinocyte growth medium without EGF or with EGF were as negative control and positive control, respectively. Cells were cultured at 2500, 37500 cells/cm 2 in dishes in serum-free medium and supernatants were collected at 24, 48 and 72 hours. The concentrations of TGF-alpha PDGF-AB and VEGF in cell culture supernatant were measured by using TGF-alpha, PDGF-AB and VEGF enzyme linked immunoadsorbent assay (ELISA) kits, respectively and cell numbers were counted by haemocytometer.


RESULTS
For cells cultured at low density and cells were subconfluent, TGF-alpha in each group was low and there was no significant difference among them at 24 hours. At 48 hours, both in absolute concentrations and on a per-cell basis, FGF-10 enhanced the secretion of TGF-alpha by cells cultured at low density in a dose-dependent fashion. At 72 hours, the concentrations of TGF-alpha in all dose of FGF-10 were significantly higher than that in negative control (P<0.05). The production of TGF-alpha in pg/10(6) cells in 500 ng/ml FGF-10 reached statistical significance compared with negative control (P<0.05). The secretion of TGF-alpha in FGF-10 were lower than that in positive control at 48 and 72 hours. For cells cuttured at 37500 cells/cm(2), the secretion of TGF-alpha stimulated by FGF-10 was similar to that when cells cuttured at lower density. PDGF-AB in each group was undetectable. When cells cuttured at high density and reached 100 percent confluent, PDGF-AB could not be detected at 24 hours. At 48 hours, both in absolute concentrations and on a per-cell basis, PDGF-AB productions in 125 and 500 ng/ml of FGF-10 were markedly greater than those of negative control at 48 and 72 hours (P<0.05), and the secretion of PDGF-AB stimulated by FGF-10 was in a dose-dependent fashion. For cells cultured at low density, although the concentrations of VEGF in 16-500 ng/ml of FGF-10 were significantly higher than that in negative control (P<0.05) at 72 hours, on a per-cell basis, they were not greater than that of negative control. The secretion of VEGF in positive control was greater than that of FGF-10 at 24-72 hours.


CONCLUSIONS
The effect of FGF-10 on increasing secretion of TGF-alpha and PDGF-AB might mediate the actions of FGF-10 on keratinocytes, fibroblasts and endothelial cells to promote re-epithelialization as well as granulation tissue deposition during wound healing.
Sun
Xiao-qing
XQ
304 th Hospital of People's Liberation Army, Beijing 100037, China.
Fu
Xiao-bing
XB
Chen
Wei
W
Sun
Tong-zhu
TZ
D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D051526:Fibroblast Growth Factor 10; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000378:metabolism; D013997:Time Factors; D015291:Transforming Growth Factors / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
C496463:FGF10 protein, human; D051526:Fibroblast Growth Factor 10; D036341:Intercellular Signaling Peptides and Proteins; D010982:Platelet-Derived Growth Factor; D042461:Vascular Endothelial Growth Factor A; C095477:platelet-derived growth factor AB; D005346:Fibroblast Growth Factors; D015291:Transforming Growth Factors