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T. Matsunaga, T. Sato, Y. Watanabe, T. Fujimaki, A. Murakami; A Device for Gene Transfer to the Anterior Segment of the Eye Using Anionic Hydrogel Contact Lens. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3114.
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To investigate non-viral device for gene transfer to the anterior segment of the eye using a newly developed anionic hydrogel soft contact lens (SCL).
We have developed a newly SCL which contained imidazolium group and phosphate group in its side chains. The anionic group of SCL retained DNA through calcium ion (SCL-DNA). The phosphate group and the DNA through calcium ion and let SCL retain DNA (SCL-DNA). Rabbit limbal epithelial cells (RLEC) were cultured onto 24-well plate and cultured with the SCL-DNA. RLEC also cultured onto the SCL-DNA. Expression of the transduced gene was monitored by fluorescence microscopy. Transfection efficiency was estimated as the percentage of GFP-positive cells identified. In vivo, rabbit corneas after wearing the SCL-DNAs were observed expression of the GFP in with a fluorescence microscope.
The retaining capacity of the DNA in the SCL was depended on the anionic content, and the SCL-DNA was released of the DNA for 24 hours or longer. In RLEC cultured with or on the SCL-DNA, expression of the GFP was observed depending on anionic content of the SCL-DNA. Furthermore, the expression of the GFP in the corneal epithelium cell which wore the DNA-SCL was observed to not only surface layer but also basal layer.
The newly developed SCL which contained imidazolium group and phosphate group in its side chains lens might be one of the efficient devices for non-viral gene transfection to corneal epithelial cells. It is suggested that this SCL could be applied for gene therapy for corneal diseases.
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