Mechanisms controlling the effects of bevacizumab (avastin) on the inhibition of early but not late formed corneal neovascularization
Authors: Fung-Rong Hu
Hsiao-Sang Chu
Lu-Ping Chow
Yan-Ming Chen
Chung-Tien Lin
Chih-Ta Chen
Wei-Li Chen
Journal: PLoS ONE (2014)
Discipline: Multidisciplinary
Keywords: bevacizumab; corneal neovascularization; early treatment; late treatment; intracorneal diffusion
Affiliations: Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan; Center of Corneal Tissue Engineering and Stem Cell Biology, National Taiwan University Hospital, Taipei, Taiwan., Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan; Department of Ophthalmology, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan., Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan., Department of Veterinary Medicine, College of Bio-Resource and Agriculture, National Taiwan University, Taipei, Taiwan., Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.

Abstract:  Purpose: To evaluate the effects and underlying mechanisms of early and late subconjunctival injection of bevacizumab on the inhibition of corneal neovascularization (NV). Methods: Corneal 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. Results: Early 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. Conclusions: Early 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. 

Retracted: No

PubPeer comments:
 Figure 5. * Red boxes: The GAPDH panel from Figure 5A (Corneal epithelium) looks very similar to the GAPDH panel from Figure 5C (Corneal stroma), albeit shown at a different exposure. Reported to the journal in October 2015, but no action taken as of today. ![file] ,