A Device for Gene Transfer to the Anterior Segment of the Eye Using Anionic Hydrogel Contact Lens

Abstract

Purpose: : 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).

Methods: : 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.

Results: : 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.

Conclusions: : 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.

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