Aim: Treatment of exudative age-related macular degeneration by using vascular endothelial growth factor (VEGF) antagonists is the gold standard today. So far, several bioactive molecules have been approved for therapeutic use. In this study, we investigate the effects of ranibizumab (Lucentis®), bevacizumab (Avastin®), and aflibercept (Eylea®) on primary human retinal pigment epithelial (hRPE) cells in vitro. Methods: hRPE cells were prepared from donor eyes and cultured under standard culture conditions. Scleral fibroblasts also prepared from donor tissue served as physiological controls. The impact of the anti-VEGF molecules on cell viability was investigated with the trypan blue exclusion assay, whereas proliferation was measured using the MTT assay. Biological activity of the molecules was quantified in a VEGF-enzyme-linked immunosorbent assay (ELISA). Results: All tested substances were biologically active in vitro. They displayed no cytotoxicity on RPE cells or scleral fibroblasts. However, proliferation of RPE cells was significantly decreased after treatment with ranibizumab or bevacizumab but not with aflibercept. Conclusions: The humanized antibodies (fragments) interfered specifically with the RPE cells. The thereby measured inhibition of cell proliferation may indicate possible side effects on the physiology of RPE cells.

1.
Pascolini D, Mariotti SP: Global estimates of visual impairment: 2010. Br J Ophthalmol 2012; 96: 614–618.
2.
Brody BL, Gamst AC, Williams RA, Smith AR, Lau PW, Dolnak D, Rapaport MH, Kaplan RM, Brown SI: Depression, visual acu ity, comorbidity, and disability associated with age-related macular degeneration. Ophthalmology 2001; 108: 1893–1900; discussion 1900–1891.
3.
Rosenfeld PJ, Brown DM, Heier JS, Boyer DS, Kaiser PK, Chung CY, Kim RY, Group MS: Ranibizumab for neovascular age-related macular degeneration. N Engl J Med 2006; 355: 1419–1431.
4.
Brown DM, Kaiser PK, Michels M, Soubrane G, Heier JS, Kim RY, Sy JP, Group AS: Rani-bizumab versus verteporfin for neovascular age-related macular degeneration. N Engl J Med 2006; 355: 1432–1444.
5.
Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA: Vascular endothelial growth factor and angiogenesis. Pharmacol Rev 2004; 56: 549–580.
6.
van Wijngaarden P, Coster DJ, Williams KA: Inhibitors of ocular neovascularization: promises and potential problems. JAMA 2005; 293: 1509–1513.
7.
Thomas M, Mousa SS, Mousa SA: Comparative effectiveness of aflibercept for the treatment of patients with neovascular age-related macular degeneration. Clin Ophthalmology 2013; 7: 495–501.
8.
Grunwald JE, Daniel E, Huang J, Ying GS, Maguire MG, Toth CA, Jaffe GJ, Fine SL, Blodi B, Klein ML, Martin AA, Hagstrom SA, Martin DF, Group CR: Risk of geographic atrophy in the comparison of age-related macular degeneration treatments trials. Ophthalmology 2014; 121: 150–161.
9.
Bhisitkul RB, Mendes TS, Rofagha S, Enanoria W, Boyer DS, Sadda SR, Zhang K: Macular atrophy progression and 7-year vision outcomes in subjects from the ANCHOR, MARINA, and HORIZON studies: the SEVEN-UP study. Am J Ophthalmol 2015; 159: 915–924 e912.
10.
Takashima A: Establishment of fibroblast cultures. Curr Protoc Cell Biol 2001, Doi 10.1002/0471143030.cb0201s00
11.
Malik D, Tarek M, Caceres del Carpio J, Ramirez C, Boyer D, Kenney MC, Kuppermann BD: Safety profiles of anti-VEGF drugs: bevacizumab, ranibizumab, aflibercept and ziv-aflibercept on human retinal pigment epithelium cells in culture. Br J Ophthalmol 2014; 98(suppl 1):i11–i16.
12.
Campa C: Effect of VEGF and anti-VEGF compounds on retinal pigment epithelium permeability: an in vitro study. Eur J Ophthalmol 2013; 23: 690–696.
13.
Klettner A, Tahmaz N, Dithmer M, Richert E, Roider J: Effects of aflibercept on primary RPE cells: toxicity, wound healing, uptake and phagocytosis. Br J Ophthalmol 2014; 98: 1448–1452.
14.
Alge CS, Suppmann S, Priglinger SG, Neubauer AS, May CA, Hauck S, Welge-Lussen U, Ueffing M, Kampik A: Comparative proteome analysis of native differentiated and cultured dedifferentiated human RPE cells. Invest Ophthalmol Vis Sci 2003; 44: 3629–3641.
15.
Ablonczy Z, Dahrouj M, Tang PH, Liu Y, Sambamurti K, Marmorstein AD, Crosson CE: Human retinal pigment epithelium cells as functional models for the RPE in vivo. Invest Ophthalmol Vis Sci 2011; 52: 8614–8620.
16.
Group CR, Martin DF, Maguire MG, Ying GS, Grunwald JE, Fine SL, Jaffe GJ: Rani-bizumab and bevacizumab for neovascular age-related macular degeneration. N Engl J Med 2011; 364: 1897–1908.
17.
Spitzer MS, Yoeruek E, Sierra A, Wallenfels-Thilo B, Schraermeyer U, Spitzer B, Bartz-Schmidt KU, Szurman P: Comparative anti-proliferative and cytotoxic profile of bevaciz-umab (Avastin), pegaptanib (Macugen) and ranibizumab (Lucentis) on different ocular cells. Graefes Arch Clin Exp Ophthalmol 2007; 245: 1837–1842.
18.
Spitzer MS, Wallenfels-Thilo B, Sierra A, Yoeruek E, Peters S, Henke-Fahle S, Bartz-Schmidt KU, Szurman P; Tuebingen Beva-cizumab Study G: Antiproliferative and cytotoxic properties of bevacizumab on different ocular cells. Br J Ophthalmol 2006; 90: 1316–1321.
19.
Brar VS, Sharma RK, Murthy RK, Chalam KV: Evaluation of differential toxicity of varying doses of bevacizumab on retinal ganglion cells, retinal pigment epithelial cells, and vascular endothelial growth factor-enriched choroidal endothelial cells. J Ocul Pharmacol Ther 2009; 25: 507–511.
20.
Puddu A, Sanguineti R, Traverso CE, Viviani GL, Nicolo M: Response to anti-VEGF-A treatment of retinal pigment epithelial cells in vitro. Eur J Ophthalmol 2016; 26: 425–430.
21.
Klettner AK, Kruse ML, Meyer T, Wesch D, Kabelitz D, Roider J: Different properties of VEGF-antagonists: bevacizumab but not ran-ibizumab accumulates in RPE cells. Graefes Arch Clin Exp Ophthalmol 2009; 247: 1601–1608.
22.
Klettner A, Mohle F, Roider J: Intracellular bevacizumab reduces phagocytotic uptake in RPE cells. Graefes Arch Clin Exp Ophthalmol 2010; 248: 819–824.
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