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Yoko Miura, Ingrid Fritz, Salvatore Grisanti, Martin Rudolf; Effect of Apolipoprotein A1-mimetic peptides on cell viability of cultured retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3296.
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© ARVO (1962-2015); The Authors (2016-present)
Previous studies of our research group have shown that ApoA-I mimetic peptides like D-4F and L-4F are able to reduce pathological lipid deposits in and on top of Bruch’s membrane (Rudolf et al. ARVO 2011, 2013). Since the function of the retinal pigment epithelium (RPE) plays a central role in photoreceptor health and function we evaluate the effect of these peptides on RPE cell viability in vitro.
Third passage porcine RPE cell cultures were treated with different concentrations of L-4F or its stereoisomer D-4F (0; 0.1; 10; 100; 500; 1000; 2000; 4000 µg/ml) for 48 hrs. Nonfunctional scrambled L-4F (sL-4F) served as a control. MTT assays (Dimethylthiazol diphenyltetrazolium bromide) were performed to analyse the peptides’ cytotoxicity on RPE cells. RPE cell proliferation activity was assessed by measuring the optical density at 577 nm using a photometric multiplate reader. Further test runs were performed in smaller concentration steps to detect the maximum tolerated dose of these peptides.
D-4F and L-4F showed no significant effect in MTT assays with concentration ≤ 10 µg/ml. With higher concentrations, L-4F ≤ 375 µg/ml and ≤ 125 µg/ml for D-4F induced a significant increase of optical density (p<0.01). L-4F concentrations ≥ 500 µg/ml reduced significantly RPE cell proliferation (p<0.01). The same effect was detected for D-4F at concentrations ≥ 250 µg/ml. The control peptide sL-4F did not show any significant difference on RPE cell proliferation activity.
Our results demonstrated cytotoxic effects in RPE cell culture if exposed to concentrations of L-4F ≥500 µg/ml and ≥ 250 µg/ml of D-4F. In RPE cell cultures concentrations up to 10 µg/ml can be considered safe for both peptides. Additional tests are needed to explain sufficiently the increase of optical density for concentrations between 10 µg/ml and the mentioned toxicity levels above.
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