June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Characterization of a Rabbit Model of Choroidal Neovascularization (CNV) and its Treatment with Lucentis®
Author Affiliations & Notes
  • Craig B Struble
    Covance Laboratories, Madison, Wisconsin, United States
  • Megan Krueger
    Covance Laboratories, Madison, Wisconsin, United States
  • Carol Rasmussen
    OSOD, LLC, Madison, Wisconsin, United States
  • Anne Goulding
    OSOD, LLC, Madison, Wisconsin, United States
  • Mike Neider
    OSOD, LLC, Madison, Wisconsin, United States
  • Hugh Wabers
    OSOD, LLC, Madison, Wisconsin, United States
  • Paul Miller
    OSOD, LLC, Madison, Wisconsin, United States
  • Ismail Zaitoun
    Ophthalmology, University of Wisconsin, Madison, Wisconsin, United States
  • Nadar Sheibani
    Ophthalmology, University of Wisconsin, Madison, Wisconsin, United States
  • Steve Van Adestine
    Covance Laboratories, Madison, Wisconsin, United States
  • Alok Sharma
    Covance Laboratories, Madison, Wisconsin, United States
  • mike Nork
    OSOD, LLC, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Craig Struble, None; Megan Krueger, None; Carol Rasmussen, None; Anne Goulding, None; Mike Neider, None; Hugh Wabers, None; Paul Miller, None; Ismail Zaitoun, None; Nadar Sheibani, None; Steve Van Adestine, None; Alok Sharma, None; mike Nork, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4076. doi:
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    • Get Citation

      Craig B Struble, Megan Krueger, Carol Rasmussen, Anne Goulding, Mike Neider, Hugh Wabers, Paul Miller, Ismail Zaitoun, Nadar Sheibani, Steve Van Adestine, Alok Sharma, mike Nork; Characterization of a Rabbit Model of Choroidal Neovascularization (CNV) and its Treatment with Lucentis®. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4076.

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

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Abstract

Purpose : To characterize an earlier CNV model in Dutch Belted rabbits (Ni, et al. 2005) using modified procedures and additional evaluation techniques.

Methods : 22 young adult Dutch Belted rabbits received subretinal injections of heparin-sepharose beads with fibroblast growth factor and LPS (100 ng bFGF, 100 ng in 50 µL of ~3% collagen gel, pH 7-7.2) in the right eye. A 2-step injection technique was used. After insertion of a 23G cannula through the pars plana, a 41G needle (Dutch Ophthalmic Research Center) was used to create a subretinal bleb with BSS followed by injection of the formulation with a blunt 26G needle through the same retinotomy. Bruch’s Membrane was perforated with a micro-vitreoretinal blade. Angiography with sodium fluorescein and indocyanin green (ICG), fundus photography, and sdOCT were conducted periodically. After angiography at Weeks 2 and 3, 16 animals were selected based on CNV lesion size and placed into 2 groups of 8 animals. Animals were dosed on Weeks 4 and 10. Groups 1 and 2 received intravitreal saline and Lucentis®, respectively. The area of the lesions visualized with ICG angiography were quantified with Image J up to 26 weeks post-lesion induction. Animals were sacrificed at 26 weeks and eyes were microscopically evaluated using H&E and Masson’s Trichrome stains, and VEGF-, B4 lectin-, and PDGF-Rβ- immune-labels.

Results : All injected eyes showed CNV lesion growth at 2 weeks continuing up to 26 weeks when the study was terminated. Treatment with Lucentis® significantly inhibited the growth of CNV lesions for up to 16 weeks as evaluated with ICG angiography (p<0.01 or p<0.05). OCT showed a thinning of the CNV lesion in the treated group. Masson’s Trichrome showed slight to minimal staining in the chorioretinal scar in both groups. VEGF immunolabeling in the chorioretinal scar occurred in the vascular endothelium of the control group, but not in the Lucentis®-treated Group. B4 lectin immunolabeling occurred more intensely in the vascular endothelium of the control group, compared to the treated group and PDGF-Rβ was observed in both groups of animals demonstrating the presence of pericytes in the CNV lesion.

Conclusions : Persistent CNV lesions were produced in Dutch Belted rabbits. The model was validated using Lucentis® as a positive control and characterized with fluorescein and ICG angiography, fundus photography, sdOCT, and histo- and immunohistochemistry

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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