Parisi Lorenzo, Fuhrer Reto, Zinkernagel Martin, Enzmann Volker
Department of Ophthalmology, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland.
Department of Biomedical Research, University of Bern, Bern, Switzerland.
Ophthalmologica. 2019;241(3):137-142. doi: 10.1159/000490430. Epub 2018 Jul 12.
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.
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).
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.
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.
使用血管内皮生长因子（VEGF）拮抗剂治疗渗出性年龄相关性黄斑变性是目前的金标准。到目前为止，已有几种生物活性分子被批准用于治疗。在本研究中，我们在体外研究了雷珠单抗（Lucentis®）、贝伐单抗（Avastin®）和阿柏西普（Eylea®）对原代人视网膜色素上皮（hRPE）细胞的影响。
从供体眼中制备hRPE细胞，并在标准培养条件下培养。同样从供体组织制备的巩膜成纤维细胞作为生理对照。用台盼蓝排斥试验研究抗VEGF分子对细胞活力的影响，而用MTT试验测量细胞增殖。在VEGF酶联免疫吸附测定（ELISA）中对分子的生物活性进行定量。
所有测试物质在体外均具有生物活性。它们对RPE细胞或巩膜成纤维细胞均无细胞毒性。然而，用雷珠单抗或贝伐单抗治疗后，RPE细胞的增殖显著降低，但用阿柏西普治疗后则没有。
人源化抗体（片段）特异性干扰RPE细胞。由此测得的细胞增殖抑制可能表明对RPE细胞生理有潜在副作用。
