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 3.973
Times Cited 3
WOS Category BIOPHYSICS MATERIALS SCIENCE, BIOMATERIALS CHEMISTRY, PHYSICAL
Resource
Human and Animal Cells