June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Hypoxia induces formation of capillaries that resemble leaky vessels in CNV
Author Affiliations & Notes
  • Marina Valerevna Tikhonovich
    Division of Experimental Vitreoretinal Surgery, Center for Ophthalmology, Tübingen, Germany
    Faculty of medicine, Lomonosov Moscow State University, Moscow, Russian Federation
  • Antje Biesemeier
    Division of Experimental Vitreoretinal Surgery, Center for Ophthalmology, Tübingen, Germany
  • Ulrich Schraermeyer
    Division of Experimental Vitreoretinal Surgery, Center for Ophthalmology, Tübingen, Germany
  • Footnotes
    Commercial Relationships   Marina Tikhonovich, None; Antje Biesemeier, None; Ulrich Schraermeyer, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4078. doi:
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    • Get Citation

      Marina Valerevna Tikhonovich, Antje Biesemeier, Ulrich Schraermeyer; Hypoxia induces formation of capillaries that resemble leaky vessels in CNV. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4078.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Ischemia of the posterior eye chamber is one of the severe problems in ophthalmology. However, little is known about changes of existing fully formed choroidal blood vessels under ischemic conditions. This study was based on a new model and it reports findings in the choroid of rats 14 hours after ischemia.

Methods : The study involved 16 adult six weeks old male rats (32 eyes). Eight pigmented and eight albino animals were used. Ischemia was modeled by incubating the eyes in PBS or DMEM at 4°C during fourteen hours after enucleation. Eyes fixed immediately after enucleation were used as a control. Transmission electron microscopy and immunohistochemical staining for HIF-1α and VEGF were performed. Statistical analysis was performed by using a nonparametric Mann-Whitney test.

Results : Retinal and choroidal cells were still alive after 14 hours of incubation at 4°C and they showed an increase of HIF-1α and VEGF compared to the control. Due to ischemia the endothelial cells of the choriocapillaris changed their morphology and made microvillar projections into the choriocapillaris lumen, which were often fenestrated. Because of the projections the area of the endothelial cell surface increased four (p<0.001) times and the length of the endothelial cell contour by 58% (p<0.001) in comparison to the control group. The endothelial cell projections tried to build up new lumina inside the existing choriocapillaris. In severe cases, the projections formed a labyrinth structure which nearly closed the vessel lumen. In addition, a lot of gaps between the endothelial cells were found. In the choroidal melanocytes of pigmented rats, a great number of degrading melanosomes were found after 14 hours of ischemia.

Conclusions : By using this easy, fast and cheap model of ischemia the first steps of choroidal neovascularization (CNV) were shown: the increase of synthesis of HIF-1α and VEGF which is very likely responsible for the formation of gaps between endothelial cells and the formation of fenestrated endothelial cell projections into the vessel lumen. Their abundance can block perfusion of blood cells and thereby inhibit closure of the endothelial gaps by platelets, but still allow the flow of plasma. This may be the reason for a permanent leakage of fluid as it was found in so-called labyrinth capillaries in surgically excised human CNV membranes before (DOI 10.1007/s00417-014-2733-0).

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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