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Impressum
Effects of advanced glycation endproducts and the cytokines bFGF/VEGF on the tube formation of choroidal endothelial cells
1Wang Y. S., 2Friedrichs U., 2Eichler W., 2Hoffmann S.; 2Wiedemann P.
2Department of Ophthalmology, Faculty of Medicine, University of Leipzig, Germany; 1Recipient of Alexander von Humboldt Foundation Fellowship, from Department of Ophthalmology, Xijing Hospital, Fourth Military Medical University, Xi'an, People's Republic of China
Objective: Age-related macular degeneration (AMD) is a leading
cause of blindness in the elderly. Advanced glycated endproducts (AGE)
are suspected to be involved in the pathogenesis of exsudative AMD. The
increased expression of AGE has been shown previously by immunohistological
methods in membranes from AMD patients. In this study we evaluated the
effects of AGE and the cytokines bFGF and VEGF on choroidal endothelial
cell (CEC) tube formation in vitro.
Methods: Bovine CEC were isolated with lycopersicion esculentum
agglutinin coated Dynabeads and cultured in EGM medium. Subconfluent cells
were used for the tube formation assay. The CEC (100,000 cells/well) were
seeded in a Vitrogen collagen matrix and stimulated with AGE (50- 500
µg/ml), VEGF (50 ng/ml) or bFGF (50 ng/ml). After a time course
of 3 days, photographs were made and digitalized. Tube formation was evaluated
from the digitalized pictures by help of the KS300 software program.
Results: The cytokines VEGF and bFGF enhanced strongly the tube
formation of CEC in a collagen matrix. In this condition, stimulation
of CEC with 50 ng/ml bFGF resulted in about 1.2-fold increase in the tube
length over the basal level (P<0.05). The AGE showed no effect on CEC
tube formation (P>0.05).
Conclusion: VEGF and bFGF are strong inducers of CEC tube formation
in vitro. Despite their described effects on the upregulation of angiogenic
cytokines in retinal endothelial/epithelial cells, AGE apparently have
no direct effect on CEC tube formation. Therefore, further studies with
AGE prestimulated retinal pigment epithelial cells and CEC in a Vitrogen
tube formation assay are needed to investigate indirect effects of AGE
on CEC tube formation.
Support: Deutsche Forschungsgemeinschaft, WI 880\9-1