Ben-Arzi Assaf, Spector Itay, Keshet Yariv, Gal-Or Orly, Bahar Irit, Dotan Assaf
Department of Ophthalmology, Rabin Medical Center, Petach Tikva, Israel.
Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Invest Ophthalmol Vis Sci. 2025 Jun 2;66(6):1. doi: 10.1167/iovs.66.6.1.
Sustained increase in intraocular pressure (IOP) following intravitreal injection of anti-vascular endothelial growth factor (anti-VEGF) in the treatment of retinal disease has been theoretically attributed to aggregation of anti-VEGF in the iridocorneal angle. However, previous studies by our group showed full clearance of intravitreally injected bevacizumab, aflibercept, and ranibizumab. The objective of this study was to further analyze and compare the clearance of these anti-VEGF agents from the eye after a single injection in a rat model.
Brown Norway rats received an intravitreal injection of 3 µl anti-VEGF at the standard concentration 3 days following induction of choroidal neovascularization. The eyes were processed at 0, 3, 6, 24, and 48 hours thereafter, and immunofluorescence was evaluated with confocal microscopy and 3D reconstruction analysis. The signal concentration was calculated, and the drug clearance rate was measured. The immunohistochemistry process was validated with negative and positive control groups.
The immunofluorescent signal was positive for all anti-VEGF agents at the trabecular meshwork, Schlemm's canal, and episcleral veins. Anti-VEGF immunostaining peaked immediately after injection and then decreased gradually to negligible at 48 hours (P < 0.05). All three agents demonstrated an identical pattern (P > 0.05). The clearance rate of anti-VEGF from the iridocorneal angle ranged between 98.68% and 99.87% at 48 hours.
Bevacizumab, aflibercept, and ranibizumab cleared completely from the iridocorneal angle at 48 hours in the brown Norway rats following a single intravitreal injection and with similar a clearance rate. These findings support our earlier studies refuting the anti-VEGF aggregation hypothesis.
在视网膜疾病治疗中，玻璃体内注射抗血管内皮生长因子（anti-VEGF）后眼压持续升高，理论上归因于抗VEGF在虹膜角膜角的聚集。然而，我们团队之前的研究表明，玻璃体内注射的贝伐单抗、阿柏西普和雷珠单抗可完全清除。本研究的目的是在大鼠模型中进一步分析和比较单次注射后这些抗VEGF药物从眼内的清除情况。
在诱导脉络膜新生血管形成3天后，给挪威棕色大鼠玻璃体内注射3 μl标准浓度的抗VEGF。此后在0、3、6、24和48小时对眼睛进行处理，并用共聚焦显微镜和三维重建分析评估免疫荧光。计算信号浓度，并测量药物清除率。免疫组化过程用阴性和阳性对照组进行验证。
在小梁网、施莱姆管和巩膜静脉处，所有抗VEGF药物的免疫荧光信号均为阳性。抗VEGF免疫染色在注射后立即达到峰值，然后逐渐下降，在48小时时降至可忽略不计（P<0.05）。所有三种药物表现出相同的模式（P>0.05）。48小时时，抗VEGF从虹膜角膜角的清除率在98.68%至99.87%之间。
在挪威棕色大鼠单次玻璃体内注射后，贝伐单抗、阿柏西普和雷珠单抗在48小时时从虹膜角膜角完全清除，清除率相似。这些发现支持了我们早期反驳抗VEGF聚集假说的研究。
