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.2003 Apr;20(4):576-83.
doi: 10.1023/a:1023238530504.

Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers

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Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers

Leena Pitkänen et al. Pharm Res.2003 Apr.

Abstract

Purpose: To investigate the role of vitreous in nonviral gene delivery into retinal pigment epithelial (RPE) cells.

Methods: Human RPE cell line D407 was cultured in six-well plates. Bovine vitreous, hyaluronan, or DMEM was added on the cells. Complexes of DNA and cationic carriers (polyethyleneimine, poly-L-lysine, DOTAP liposomes) were pipetted onto the vitreous, hyaluronan, or DMEM. Cellular uptake of DNA was studied with ethidium monoazide DNA and gene expression with GFP-plasmid complexes. FITC-dextrans and FITC-polylysines were used to probe the effects of the size and cationic charge on permeation in the vitreous in a similar experimental setup. Fluorescent cells were analyzed by flow cytometry.

Results: Vitreous decreased the cellular uptake of DNA complexes 2-30 times, and GFP expression was also impaired. In hyaluronan solutions the cellular uptake of the complexes was also decreased significantly in most cases. In vitreous, cellular uptake of all FITC-dextrans decreased slightly, and uptake of poly-L-lysines was decreased substantially, whereas in hyaluronan solutions the effects were mild or nonexistent.

Conclusions: Polymeric and liposomal gene delivery is substantially limited by the vitreous. This is probably because of the size and charge of the retinal gene delivery after intravitreal injections.

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