The effects of growth factors on the proliferation and in vitro angiogenesis of human macular inner choroidal endothelial cells.
OBJECTIVE
To investigate the effect of VEGF(165), FGF2, IGF-1, PDGF-AA, PDGF-BB and IL-1 beta on the proliferation and angiogenic tube formation of human macular inner choroidal endothelial cells (ICEC).


METHODS
The proliferation of human macular ICECs after exposure to the aforementioned growth factors was determined by using both a WST-1 colorimetric assay and a cell-counting technique. The effect of growth factors on ICEC angiogenesis was assessed by sprout formation using a three-dimensional in vitro Matrigel duplex assay.


RESULTS
Using both the WST-1 assay and a cell-counting technique, VEGF(165) and FGF2 both significantly increased human macular ICEC proliferation. The effect of equimolar concentrations of VEGF(165) and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1 beta on proliferation up to a growth factor concentration of 1000 pmol/l. The angiogenesis assay found a significant effect on sprout formation for VEGF(165) and FGF-2. Again, the effect of equimolar concentrations of VEGF(165) and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1 beta on sprout formation at 1000 pmol/l.


CONCLUSIONS
Both VEGF(165) and FGF2 significantly increase human macular ICEC proliferation and sprout formation in an angiogenesis assay. When present together, their effect was additive. IGF-1, PDGF-AA, PDGF-BB and IL-1 beta did not have any significant effect on proliferation or sprout formation in vitro. These results suggest that targeting other growth factors such as FGF2, in addition to VEGF, may be beneficial in the treatment of neovascular age-related macular degeneration.
Browning
A C
AC
Division of Ophthalmology and Visual Sciences, Eye, Ear, Nose and Throat Centre, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK.
Dua
H S
HS
Amoaku
W M
WM
D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002829:Choroid / Q000166:cytology / Q000187:drug effects*; D020256:Choroidal Neovascularization / Q000473:pathology*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D006133:Growth Substances; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2