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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.
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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.
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.
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.
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.
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