RRC ID |
45221
|
Author |
Yazaki Y, Oyane A, Tsurushima H, Araki H, Sogo Y, Ito A, Yamazaki A.
|
Title |
Improved gene transfer efficiency of a DNA-lipid-apatite composite layer by controlling the layer molecular composition.
|
Journal |
Colloids Surf B Biointerfaces
|
Abstract |
Surface-mediated nonviral gene transfer systems using biocompatible apatite-based composite layers have potential use in tissue engineering applications. Herein, we investigated a relatively efficient system based on a DNA-lipid-apatite composite layer (DLp-Ap layer): an apatite (Ap) layer with immobilized DNA and lipid (Lp) complexes (DLp complexes). DLp-Ap layers were fabricated on substrates using supersaturated calcium phosphate coprecipitation solutions supplemented with DLp complexes, and the molecular compositions of the DLp-Ap layers were controlled by varying the net DNA concentrations and Lp/DNA ratios in the coprecipitation solutions. Increases in both the DNA concentration and Lp/DNA ratio in the coprecipitation solution increased the DLp complex content of the resulting DLp-Ap layer. However, a higher DLp complex content did not always provide increased gene transfer efficiency to the CHO-K1 cells, because there was a threshold content of approximately 10μg/cm(2). In addition, DLp-Ap layers with similar DLp complex contents exhibited different gene transfer efficiencies, most likely due to the different Lp/DNA ratios in the layers. Notably, the optimized Lp/DNA ratios in the coprecipitation solutions for maximizing the gene transfer efficiency were lower than those of the conventional particle-mediated lipofection systems. These findings will serve as a useful design guide for the preparation of DLp-Ap layers with high gene transfer efficiency.
|
Volume |
122
|
Pages |
465-471
|
Published |
2014-10-1
|
DOI |
10.1016/j.colsurfb.2014.07.001
|
PII |
S0927-7765(14)00363-4
|
PMID |
25096721
|
MeSH |
Apatites / chemistry*
DNA / chemistry*
Gene Transfer Techniques*
Lipids / chemistry*
Microscopy, Electron, Scanning
|
IF |
4.389
|
Times Cited |
3
|
WOS Category
|
BIOPHYSICS
MATERIALS SCIENCE, BIOMATERIALS
CHEMISTRY, PHYSICAL
|
Resource |
Human and Animal Cells |
CHO-K1 |