Department of Ophthalmology, University of Cologne, Cologne, Germany.
Invest Ophthalmol Vis Sci. 2013 Sep 11;54(9):6133-42. doi: 10.1167/iovs.13-12460.
The effect of autologous serum eyedrops on the corneal vasculature is undefined. Therefore, we analyzed the corneal vascular effects of serum eyedrops in comparison with and in combination with bevacizumab eyedrops.
In vitro, blood and lymphatic endothelial cells were treated with serum eyedrops, bevacizumab eyedrops, or a combination of both, and cell proliferation was measured. In vivo, inflammatory corneal neovascularization was induced by suture placement. Subsequently, corneal blood and lymphatic vessel progression and regression were analyzed after treatment with serum or bevacizumab eyedrops or a combination of both. Hemangiogenesis and lymphangiogenesis were quantified in whole mounts using CD31 and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1); inflammatory cell infiltration was analyzed using CD11b. Furthermore, corneal expression levels of interleukin 1β, tumor necrosis factor α, vascular endothelial growth factor (VEGF) A, VEGF-C, and VEGF-D were analyzed by real-time PCR.
In vitro, serum increased and bevacizumab decreased endothelial cell proliferation. In vivo, serum eyedrops had no significant effect on corneal vessel progression or regression. Bevacizumab eyedrops reduced blood and lymphatic vessel progression and promoted blood and lymphatic vessel regression. The combination of serum eyedrops and bevacizumab eyedrops attenuated the anti(lymph)angiogenic effects of bevacizumab. Inflammatory corneal cell counts were not significantly altered by serum or bevacizumab eyedrops. Serum eyedrops changed the proinflammatory and pro(lymph)angiogenic status of the cornea. Bevacizumab eyedrops significantly reduced proinflammatory and pro(lymph)angiogenic factor expression. Higher doses of bevacizumab did not restore its anti(lymph)angiogenic effects when used in combination with serum.
The counteracting effects of serum eyedrops and bevacizumab eyedrops on the corneal vasculature should be taken into account when combined therapeutic regimens are considered.
自体血清滴眼液对角膜血管的影响尚不清楚。因此，我们分析了血清滴眼液与贝伐单抗滴眼液联合应用对角膜血管的影响。
在体外，用血清滴眼液、贝伐单抗滴眼液或两者联合处理血液和淋巴管内皮细胞，并测量细胞增殖情况。在体内，通过缝线放置诱导炎症性角膜新生血管形成。随后，用血清或贝伐单抗滴眼液或两者联合处理后，分析角膜血管的进展和消退。使用 CD31 和淋巴管内皮透明质酸受体 1（LYVE-1）对整个角膜进行血管生成和淋巴管生成的定量分析；用 CD11b 分析炎症细胞浸润。此外，通过实时 PCR 分析角膜中白细胞介素 1β、肿瘤坏死因子 α、血管内皮生长因子（VEGF）A、VEGF-C 和 VEGF-D 的表达水平。
在体外，血清增加而贝伐单抗减少内皮细胞增殖。在体内，血清滴眼液对角膜血管的进展或消退没有显著影响。贝伐单抗滴眼液可减少血管和淋巴管的进展，并促进血管和淋巴管的消退。血清滴眼液和贝伐单抗滴眼液的联合使用可减弱贝伐单抗的抗（淋）血管生成作用。血清或贝伐单抗滴眼液对炎症性角膜细胞计数没有显著影响。血清滴眼液改变了角膜的促炎和促（淋）血管生成状态。贝伐单抗滴眼液显著降低了促炎和促（淋）血管生成因子的表达。当与血清联合使用时，较高剂量的贝伐单抗并不能恢复其抗（淋）血管生成作用。
在考虑联合治疗方案时，应考虑血清滴眼液和贝伐单抗滴眼液对角膜血管的拮抗作用。
