A novel in vivo model of puncture-induced iris neovascularization

PLoS One. 2017 Jun 28;12(6):e0180235. doi: 10.1371/journal.pone.0180235. eCollection 2017.

Abstract

Purpose: To assess iris neovascularization by uveal puncture of the mouse eye and determine the role of angiogenic factors during iris neovascularization.

Methods: Uveal punctures were performed on BalbC mouse eyes to induce iris angiogenesis. VEGF-blockage was used as an anti-angiogenic treatment, while normoxia- and hypoxia-conditioned media from retinal pigment epithelium (RPE) cells was used as an angiogenic-inducer in this model. Iris vasculature was determined in vivo by noninvasive methods. Iris blood vessels were stained for platelet endothelial cell adhesion molecule-1 and vascular sprouts were counted as markers of angiogenesis. Expression of angiogenic and inflammatory factors in the puncture-induced model were determined by qPCR and western blot.

Results: Punctures led to increased neovascularization and sprouting of the iris. qPCR and protein analysis showed an increase of angiogenic factors, particularly in the plasminogen-activating receptor and inflammatory systems. VEGF-blockage partly reduced iris neovascularization, and treatment with hypoxia-conditioned RPE medium led to a statistically significant increase in iris neovascularization.

Conclusions: This study presents the first evidence of a puncture-induced iris angiogenesis model in the mouse. In a broader context, this novel in vivo model of neovascularization has the potential for noninvasive evaluation of angiogenesis modulating substances.

MeSH terms

  • Animals
  • Blotting, Western
  • Disease Models, Animal
  • Female
  • Fluorescent Antibody Technique
  • Iris / blood supply
  • Iris / injuries*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Neovascularization, Pathologic / etiology*
  • Neovascularization, Pathologic / pathology
  • Real-Time Polymerase Chain Reaction
  • Vascular Endothelial Growth Factor A / antagonists & inhibitors
  • Wounds, Penetrating / pathology*

Substances

  • Vascular Endothelial Growth Factor A

Grants and funding

The presented work was supported by grants from the Regnérs Foundation for Research in Eye Diseases, the Swedish Eye Foundation, the Cronqvist Foundation, and the Karolinska Institutet; Ophélie Beauhean was partly supported by an Erasmus Trainee scholarship.