April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Gene Transfer to the Corneal Epithelial Cells Using a Hydrogel Contact Lens
Author Affiliations & Notes
  • T. Matsunaga
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
    Research and Development, SEED Co. Ltd., Saitama, Japan
  • T. Sato
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
    Research and Development, SEED Co. Ltd., Saitama, Japan
  • Y. Watanabe
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
    Research and Development, SEED Co. Ltd., Saitama, Japan
  • H. Toshida
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
  • T. Fujimaki
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
  • A. Murakami
    Department of Ophthalmology, Juntendo University, Tokyo, Japan
  • Footnotes
    Commercial Relationships  T. Matsunaga, None; T. Sato, None; Y. Watanabe, None; H. Toshida, None; T. Fujimaki, None; A. Murakami, None.
  • Footnotes
    Support  Grants-in-Aid for Scientific Research from Japan Society for the Promotion of Science #20592061
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1733. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      T. Matsunaga, T. Sato, Y. Watanabe, H. Toshida, T. Fujimaki, A. Murakami; Gene Transfer to the Corneal Epithelial Cells Using a Hydrogel Contact Lens. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1733.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : The aim of the present study was to investigate non-viral gene transfection into the corneal tissues using a newly developed hydrogel contact lens both in vitro and in vivo.

Methods: : We have developed a new contact lens from a polymer gel which contained phosphate group in its side chains. A plasmid DNA which expressed Green Fluorescent Protein (GFP) was used for reporter gene. The plasmid DNA was absorbed and retained in the gel through the formation of calcium-phosphate-DNA complex. In vitro, release behavior of the DNA from the lens was measured with a microplate reader. Human corneal epithelial cells (HCE-T) were cultured on the contact lens. Transfection efficiency and expression of GFP was monitored by a fluorescence microscope. In vivo,the contact lenses were worn on rabbit eyes, GFP expression was also observed with a fluorescence microscope.

Results: : The retaining capacity of calcium was increased according to the content of phosphate groups in the gel. The retaining of the DNA in the gel was depended on the phosphate group in a dose-dependent manner; the gel containing 10% phosphate group retained the DNA twofold than 5% phosphate group. Furthermore, sustained release of plasmid DNA from the gel with 10% phosphate group was observed for three days or longer. GFP expression was observed both in HCE-T in vitro and rabbit corneal epithelium in vivo.

Conclusions: : The newly developed hydrogel contact lens might be one of the efficient devices for non-viral gene transfection to the corneal epithelial cells. It is suggested that this lens could be applied for gene therapy for corneal diseases.

Keywords: gene transfer/gene therapy • cornea: epithelium • contact lens 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×