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Effect of PEDF in Combination with Growth-factors on Proliferation of Bovine Retinal Endothelial Cells
1Baldysiak-Figiel A., 2Kampmeier J., 2Lang G. K., 2Lang G. E.,
1Universität Ulm, Augenklinik und Poliklinik, Augenlabor (Ulm)
2Universität Ulm, Augenklinik und Poliklinik (Ulm)
Purpose: Proliferative retinopathies such as diabetic retinopathy or age-related macular degeneration account for major causes of blindness in industrialized countries. One of the possible targets in the therapy of these disorders is neovascularization, induced by growth factors. Recently, the novel serine protease inhibitor PEDF was shown to have strong antiproliferative effects on retinoblastoma cells. In this study we investigated if PEDF alters the effects of IGF-1, VEGF, bFGF and TGF ß2 on proliferation of bovine retinal microvascular endothelial cells (BREC).
Method: Initially the maximum effective stimulatory and inhibitory concentration of PEDF was determined. The medium on confluent cell cultures was replaced with serum free medium for 24 h and then kept for another 24 h with IGF-1 (50ng/ml), bFGF (10ng/ml), VEGF (20ng/ml), TGF ß2 (50ng/ml) or PEDF (1ng/ml) alone or in combination of the growth factors with PEDF. 3H-thymidine incorporation as a direct measure of BREC proliferation was assessed by liquid scintillation counting. In all experiments untreated serum free negative controls were used. Statistical analysis was performed using the two sample t-test.
Results: The proliferation of BREC significantly increased (p=0.03) in the presence of PEDF at the concentration of 100ng/ml. PEDF directly inhibited the proliferation of BREC at the concentration of 1ng/ml (p=0.003). As compared to untreated control PEDF at the concentration of 1ng/ml showed significant inhibition of bFGF, VEGF and IGF-1 induced proliferation (p<0.0001). Moreover, the combination of PEDF (1ng/ml) and TGF ß2 (50ng/ml) had a strong synergistic antiproliferative effect (p<0,0001).
Conclusions: Our data show that PEDF alone has a concentration-dependent modulatory effect on the proliferation of bovine retinal microvascular endothelial cells. PEDF is able to inhibit the proliferation of endothelial cells induced by VEGF, IGF-1 and bFGF in vitro. Additionally, PEDF acts synergistically with TGF ß2 towards inhibition of BREC proliferation. This data show that the anti-proliferative properties of PEDF can be of possible therapeutic value for proliferative retinopathies.