Spitzer Martin S, Yoeruek Efdal, Sierra Ana, Wallenfels-Thilo Barbara, Schraermeyer Ulrich, Spitzer Bernhard, Bartz-Schmidt Karl U, Szurman Peter
University Eye Clinic Tuebingen, Department I, Eberhard-Karls University Tuebingen, Schleichstr. 12, 72076 Tuebingen, Germany.
Graefes Arch Clin Exp Ophthalmol. 2007 Dec;245(12):1837-42. doi: 10.1007/s00417-007-0568-7. Epub 2007 Mar 9.
To compare the antiproliferative and cytotoxic properties of bevacizumab (Avastin), pegaptanib (Macugen) and ranibizumab (Lucentis) on human retinal pigment epithelium (ARPE19) cells, rat retinal ganglion cells (RGC5) and pig choroidal endothelial cells (CEC).
Monolayer cultures of ARPE19, RGC5 and CEC were used. Bevacizumab (0.1-0.3 mg/ml), pegaptanib (0.025-0.08 mg/ml) or ranibizumab (0.04-0.125 mg/ml) diluted in culture medium were added to the cells. Expression of VEGF-receptors (VEGFR1 and VEGFR2) and von Willebrand factor (a marker for endothelial cells) were analysed by immunohistochemistry. CEC cells were stimulated with VEGF. Cellular proliferative activity was monitored by BrdU-incorporation into cellular DNA. For cytotoxicity assays cells were grown to confluence and then cultured in a serum-depleted medium to ensure a static milieu. MTT-test was performed after one day.
CEC and ARPE19 cells stained positively for VEGFR1 and VEGFR2. More than 95% of the CEC cells were positive for von Willebrand factor. Ranibizumab reduced CEC cell proliferation by 44.1%, bevacizumab by 38.2% and pegaptanib by 35.1% when the drugs were used at their established clinical doses. The differences, however, between the three drugs in respect to cell growth inhibition were not statistically significant. Only a mild antiproliferative effect of bevacizumab or pegaptanib on ARPE19 cells could be observed. Ranibizumab did not alter ARPE19 cell proliferation. No cytotoxicity on RGC5, CEC and ARPE19 cells could be seen.
Bevacizumab, pegaptanib and ranibizumab significantly suppress choroidal endothelial cell proliferation. However, when used at the currently established doses none of the drugs was superior over the others in respect to endothelial cell growth inhibition. The biocompatibility of all three drugs--including the off-label bevacizumab--seems to be excellent when used at the currently recommended intravitreal dose.
比较贝伐单抗（阿瓦斯汀）、培加替尼（麦考真金）和雷珠单抗（ Lucentis）对人视网膜色素上皮（ARPE19）细胞、大鼠视网膜神经节细胞（RGC5）和猪脉络膜内皮细胞（CEC）的抗增殖和细胞毒性特性。
使用ARPE19、RGC5和CEC的单层培养物。将在培养基中稀释的贝伐单抗（0.1 - 0.3 mg/ml）、培加替尼（0.025 - 0.08 mg/ml）或雷珠单抗（0.04 - 0.125 mg/ml）添加到细胞中。通过免疫组织化学分析VEGF受体（VEGFR1和VEGFR2）和血管性血友病因子（内皮细胞标志物）的表达。用VEGF刺激CEC细胞。通过将BrdU掺入细胞DNA中来监测细胞增殖活性。对于细胞毒性测定，将细胞培养至汇合，然后在无血清培养基中培养以确保静态环境。一天后进行MTT试验。
CEC和ARPE19细胞对VEGFR1和VEGFR2染色呈阳性。超过95%的CEC细胞对血管性血友病因子呈阳性。当以既定临床剂量使用药物时，雷珠单抗使CEC细胞增殖减少44.1%，贝伐单抗减少38.2%，培加替尼减少35.1%。然而，三种药物在细胞生长抑制方面的差异无统计学意义。仅观察到贝伐单抗或培加替尼对ARPE19细胞有轻微的抗增殖作用。雷珠单抗未改变ARPE19细胞增殖。未观察到对RGC5、CEC和ARPE19细胞的细胞毒性。
贝伐单抗、培加替尼和雷珠单抗可显著抑制脉络膜内皮细胞增殖。然而，当以目前既定剂量使用时，在抑制内皮细胞生长方面，没有一种药物优于其他药物。当以目前推荐的玻璃体内剂量使用时，所有三种药物（包括未按标签使用的贝伐单抗）的生物相容性似乎都很好。
