Ari Seyhmus, Nergiz Yusuf, Aksit Ihsan, Sahin Alparslan, Cingu Kursat, Caca Ihsan
1 Department of Ophthalmology, Dicle University Medical School , Diyarbakir, Turkey .
J Ocul Pharmacol Ther. 2015 Mar;31(2):100-5. doi: 10.1089/jop.2014.0005. Epub 2015 Jan 9.
To evaluate the effects of intracameral injection of ranibizumab and bevacizumab on the corneal endothelium by scanning electron microscopy (SEM).
Twenty-eight female rabbits were randomly divided into four equal groups. Rabbits in groups 1 and 2 underwent intracameral injection of 1 mg/0.1 mL and 0.5 mg/0.05 mL ranibizumab, respectively; group 3 was injected with 1.25 mg/0.05 mL bevacizumab. All three groups were injected with a balanced salt solution (BSS) into the anterior chamber of the left (fellow) eye. None of the rabbits in group 4 underwent an injection. Corneal thickness and intraocular pressure were measured before the injections, on the first day, and in the first month after injection. The rabbits were sacrificed and corneal tissues were excised in the first month after injection. Specular microscopy was used for the corneal endothelial cell count. Endothelial cell density was assessed and comparisons drawn between the groups and the control. Micrographs were recorded for SEM examination. The structure of the corneal endothelial cells, the junctional area of the cell membrane, the distribution of microvillus, and the cell morphology of the eyes that underwent intracameral injection of vascular endothelial growth factor (VEGF), BSS, and the control group were compared.
Corneal thickness and intraocular pressure were not significantly different between the groups that underwent anti-VEGF or BSS injection and the control group on the first day and in the first month of injection. The corneal endothelial cell count was significantly diminished in all three groups; predominantly in group 1 and 2 (P<0.05). The SEM examination revealed normal corneal endothelial histology in group 3 and the control group. Eyes in group 1 exhibited indistinctness of corneal endothelial cell borders, microvillus loss in the luminal surface, excessive blebbing, and disintegration of intercellular junctions. In group 2, the cell structure of the corneal endothelium and intercellular junctions were normal. However, a relative reduction was observed in the microvillus density of endothelial cells. Although eyes in group 3 were morphologically similar to fellow eyes and the control group, disarrangement in endothelial cell borders was evident.
The SEM examination pointed out deterioration in endothelial cell morphology after intracameral injection of 1 and 0.5 mg ranizumab. However, the effects of intracameral bevacizumab injection on corneal endothelial cells were similar to those found in fellow eyes and the control group. Further large-scale studies that examine the cellular changes by transmission electron microscopy are required to support the results of the present study that evaluates the structural changes in endothelial cells by SEM.
通过扫描电子显微镜(SEM)评估前房注射雷珠单抗和贝伐单抗对角膜内皮的影响。
28只雌性兔子随机分为四组,每组数量相等。第1组和第2组兔子分别前房注射1mg/0.1mL和0.5mg/0.05mL雷珠单抗;第3组注射1.25mg/0.05mL贝伐单抗。所有三组均向左眼(对侧眼)前房注射平衡盐溶液(BSS)。第4组兔子均未接受注射。在注射前、注射第一天及注射后第一个月测量角膜厚度和眼压。在注射后第一个月处死兔子并切除角膜组织,用镜面显微镜进行角膜内皮细胞计数,评估内皮细胞密度并在各实验组与对照组之间进行比较,记录显微照片用于SEM检查,比较前房注射血管内皮生长因子(VEGF)、BSS的实验组及对照组兔眼的角膜内皮细胞结构、细胞膜连接区、微绒毛分布及细胞形态。
抗VEGF或BSS注射组与对照组在注射第一天及第一个月时角膜厚度和眼压无显著差异。所有三组的角膜内皮细胞计数均显著减少,主要是第1组和第2组(P<0.05)。SEM检查显示第3组和对照组角膜内皮组织学正常。第1组兔眼角膜内皮细胞边界模糊,腔面微绒毛缺失,大量气泡形成,细胞间连接解体。第2组角膜内皮细胞结构和细胞间连接正常,但内皮细胞微绒毛密度相对降低。虽然第3组兔眼形态与对侧眼及对照组相似,但内皮细胞边界排列紊乱。
SEM检查指出前房注射1mg和0.5mg雷珠单抗后内皮细胞形态恶化。然而,前房注射贝伐单抗对角膜内皮细胞的影响与对侧眼及对照组相似。需要进一步通过透射电子显微镜检查细胞变化的大规模研究来支持本研究通过SEM评估内皮细胞结构变化的结果。