Mechanisms Controlling the Effects of Bevacizumab (Avastin) on the Inhibition of Early but Not Late Formed Corneal Neovascularization

Abstract

PurposeTo evaluate the effects and underlying mechanisms of early and late subconjunctival injection of bevacizumab on the inhibition of corneal neovascularization (NV).MethodsCorneal NV was induced by closed eye contact lens wear followed by a silk suture tarsorrhaphy in rabbits. Weekly subconjunctival injections of bevacizumab (5.0 mg) for 1 month were started immediately (early treatment group) or 1 month after induction of corneal NV with continuous induction (late treatment group). The severity of corneal NV was evaluated. Immunostaining was used to evaluate the intracorneal diffusion of bevacizumab, and the existence of pericytes and smooth muscle cells around the NV. The expression of AM-3K, an anti-macrophage antibody, vascular endothelial growth factor (VEGF) with its receptors (VEGFR1 and VEGFR2), and vascular endothelial apoptosis were also evaluated. Western blot analysis was performed to quantify the expression level of VEGF, VEGFR1 and VEGFR2 on corneal epithelium and stroma in different groups.ResultsEarly treatment with bevacizumab inhibited corneal NV more significantly than late treatment. Intracorneal diffusion of bevacizumab was not different among different groups. Immunostaining showed pericytes and smooth muscle cells around newly formed vessels as early as 2 weeks after induction. Immunostaining and Western blot analysis showed that VEGF, VEGFR1, and VEGFR2 on corneal stroma increased significantly in no treatment groups and late treatment groups, but not in early treatment group. Bevacizumab significantly inhibited macrophage infiltration in the early but not late treatment group. Sporadic vascular endothelial apoptosis was found at 4 weeks in the late but not early treatment group.ConclusionsEarly but not late injection of bevacizumab inhibited corneal NV. Late injection of bevacizumab did not alter macrophage infiltration, and can't inhibit the expression of VEGF, VEGFR1, and VEGFR2 on corneal vessels. The inhibition of corneal NV in early treatment group does not occur via vascular endothelial apoptosis.

Recent drug therapies for corneal neovascularization

“…Late treatment with bevacizumab did not suppress macrophage infiltration. It could not inhibit the expression of VEGF, VEGFR1, and VEGFR2 on corneal vessels either . Hamdan et al.…”

Regulation of matrix metalloproteinases 2 and 9 in corneal neovascularization

“…As far as we know, some drugs have been confirmed to possess the capacity for inhibition of CNV, such as bevacizumab, triamcinolone acetonide and dexamethasone which can be taken to treat CNV, are partially effective and with some side-effects (Chen et al, 2014;Choksi et al, 2013;Mehrjardi, Ghaffari, Mahbod, & Hashemi, 2014;Papathanassiou, Theodoropoulou, Analitis, Tzonou, & Theodossiadis, 2013). Currently, there is no specific and approved drug for the treatment of CNV (Menzel-Severing, 2012).…”

Mechanisms of corneal neovascularization and modern options for its suppression

“…Некоторые случаи демонстрируют резистентность к анти-VEGF-терапии. Причинами рефрактерности к лечению может быть применение анти-VEGF препаратов в поздних фазах ангиогенеза, когда просвет сосуда уже сформирован и перициты создают плотные контакты, в результате потребность в VEGF для функционирования сосудов снижается [62,63]. Другим механизмом рефрактерности может быть регуляция ангиогенеза через альтернативные пути посредством активации других стимуляторов ангиогенеза [64].…”