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In vivo efficacy of Vmb-9 in suppressing corneal neovascularization after alkali injury. ( A ) Representative images from BALB/cJ mice subjected to corneal alkali burn and treated with vehicle or Vmb-9 are shown at days 0, 1, 3, and 7 post-injury. Vehicle-treated eyes display progressive corneal neovascularization, whereas Vmb-9-treated eyes show marked attenuation of vascular ingrowth over time. For each time point, fluorescein staining ( top ) highlights epithelial integrity, ring light images ( middle ) document gross cornea integrity, and white-light images ( bottom ) show overall corneal clarity. Arrows indicated neovascularization in untreated mice. Right , Corneal whole-mount immunostaining for CD31 (blood vessels) and LYVE1 (lymphatic vessels) confirms a reduction in both hemangiogenesis and lymphangiogenesis in Vmb-9-treated corneas. ( B, C ) Quantitative analysis of CD31 + B and LYVE1 + C was performed using skeletonized images and analyzed with the Analyze Skeleton (2D/3D) plugin in Fiji (ImageJ, version 2.16.0), from which network parameters including the number of branches and number of junctions were extracted for each condition.

Journal: Investigative Ophthalmology & Visual Science

Article Title: Development of Miniprotein Binders to Inhibit VEGF-Induced Corneal Neovascularization

doi: 10.1167/iovs.67.5.61

Figure Lengend Snippet: In vivo efficacy of Vmb-9 in suppressing corneal neovascularization after alkali injury. ( A ) Representative images from BALB/cJ mice subjected to corneal alkali burn and treated with vehicle or Vmb-9 are shown at days 0, 1, 3, and 7 post-injury. Vehicle-treated eyes display progressive corneal neovascularization, whereas Vmb-9-treated eyes show marked attenuation of vascular ingrowth over time. For each time point, fluorescein staining ( top ) highlights epithelial integrity, ring light images ( middle ) document gross cornea integrity, and white-light images ( bottom ) show overall corneal clarity. Arrows indicated neovascularization in untreated mice. Right , Corneal whole-mount immunostaining for CD31 (blood vessels) and LYVE1 (lymphatic vessels) confirms a reduction in both hemangiogenesis and lymphangiogenesis in Vmb-9-treated corneas. ( B, C ) Quantitative analysis of CD31 + B and LYVE1 + C was performed using skeletonized images and analyzed with the Analyze Skeleton (2D/3D) plugin in Fiji (ImageJ, version 2.16.0), from which network parameters including the number of branches and number of junctions were extracted for each condition.

Article Snippet: BALB/cJ mice aged 6 weeks were obtained from The Jackson Laboratory (strain 000651; Bar Harbor, ME, USA) and allowed to acclimate for 1 week upon arrival.

Techniques: In Vivo, Staining, Immunostaining