Premium
Combined ultrasound‐targeted microbubble destruction and polyethylenimine‐mediated plasmid DNA delivery to the rat retina: enhanced efficiency and accelerated expression
Author(s) -
Li Hongli,
Qian Jin,
Yao Chunfang,
Wan Caifeng,
Li Fenghua
Publication year - 2016
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2875
Subject(s) - polyethylenimine , retina , transfection , gene delivery , genetic enhancement , in vivo , microbiology and biotechnology , retinal , retinal pigment epithelium , transgene , biology , chemistry , gene , biochemistry , genetics , neuroscience
Background Gene therapy has potential in the treatment of refractory retinal diseases. It is important to develop an effective delivery system in the retina. The present study aimed to investigate the efficacy and safety of ultrasound (US)‐targeted microbubble destruction (UTMD)‐mediated polyethylenimine (PEI) to the rat retina. Methods Gene transfer was examined by injecting PEI/plasmid DNA (pDNA) with or without microbubbles (MBs) into the subretinal space of rats that were then exposed to US. We investigated enhanced green fluorescent protein (eGFP) expression on flat fundus oculi and performed quantitative analysis. Hematoxylin and eosin staining was used to observe tissue damage. Results UTMD significantly enhanced PEI/pDNA transfection efficiency safely by increasing both the transgene expression per cell and the percentage of transfected cells of the retina. PEI/pDNA combined with UTMD significantly increased the number of DNA gene copies and the mRNA level in the retinal pigment epithelium (RPE) and neural retina, respectively, compared to PEI/pDNA alone. Conclusions The present study demonstrates that enhanced and accelerated pDNA expression can be achieved in the retina/RPE cells in vivo by UTMD physical techniques combined with a PEI chemical vector. Our study provides useful information for further in vivo retinal gene therapy work. Copyright © 2016 John Wiley & Sons, Ltd.