Aim: To investigate the effect of VEGF165, FGF2, IGF-1, PDGF-AA, PDGF-BB and IL-1β 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, VEGF165 and FGF2 both significantly increased human macular ICEC proliferation. The effect of equimolar concentrations of VEGF165 and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1β on proliferation up to a growth factor concentration of 1000 pmol/l. The angiogenesis assay found a significant effect on sprout formation for VEGF165 and FGF-2. Again, the effect of equimolar concentrations of VEGF165 and FGF2 was additive. There was no significant effect for IGF-1, PDGF-AA, PDGF-BB or IL-1β on sprout formation at 1000 pmol/l.
Conclusions: Both VEGF165 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-1β 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.
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Funding: This work was supported by a grant from the British Eye Research Foundation (formerly the Iris Fund). The grant provider played no role in the study design, analysis, writing of this manuscript or the decision to publish this research.
Competing interests: None.
Ethics approval: The research had the approval of the local research ethics committee (Nottingham Q1060301).
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