Subconjunctival aflibercept inhibits corneal angiogenesis and VEGFR-3+CD11b+ cells.

Abstract

Purpose: This study aimed to investigate the effects of subconjunctival injection of aflibercept, a soluble protein decoy for VEGFR-1 and VEGFR-2, on corneal angiogenesis and VEGFR-expressing CD11b⁺ cells in a mouse model of suture-induced corneal neovascularization.

Methods: Corneal neovascularization was induced in BALB/c mice by placing three sutures on the cornea. Immediately after surgery, either 200 µg aflibercept (5 µL) or an equal volume of phosphate-buffered saline (PBS) was administered into the subconjunctival space. Seven days after later, corneal new vessels were quantified through clinical examination and measurement of the CD31-stained area in corneal flat mounts. The levels of pro-angiogenic and inflammatory markers in the cornea were evaluated using RT-qPCR. The percentages of VEGFR-2⁺CD11b⁺ cells and VEGFR-3⁺CD11b⁺ cells were analyzed in the cornea, blood, and draining cervical lymph nodes (DLNs) using flow cytometry.

Results: Subconjunctival injection of aflibercept significantly reduced the growth of corneal new vessels compared to subconjunctival PBS injection. The mRNA levels of Cd31, vascular growth factors (Vegfc and Angpt1), and pro-angiogenic/inflammatory markers (Tek/Tie2, Mrc1, Mrc2, and Il6) in the cornea were downregulated by subconjunctival aflibercept. Also, the percentage of VEGFR-3⁺CD11b⁺ cells in the cornea, blood, and DLNs was decreased by aflibercept, whereas that of VEGFR-2⁺CD11b⁺ cells was unaffected.

Conclusion: Subconjunctival aflibercept administration inhibits inflammatory angiogenesis in the cornea and reduces the numbers of cornea-infiltrating and circulating VEGFR-3⁺CD11b⁺ cells.