Improved gene transfer efficiency of a DNA-lipid-apatite composite layer by controlling the layer molecular composition

Yushin Yazaki, Ayako Oyane, Hideo Tsurushima, Hiroko Araki, Yu Sogo, Atsuo Ito, Atsushi Yamazaki

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

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/cm2. 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.

Original languageEnglish
Pages (from-to)465-471
Number of pages7
JournalColloids and Surfaces B: Biointerfaces
Volume122
DOIs
Publication statusPublished - 2014 Oct 1

Keywords

  • Apatite
  • Composite
  • DNA
  • Gene transfer
  • Lipid
  • Lipofection

ASJC Scopus subject areas

  • Biotechnology
  • Surfaces and Interfaces
  • Physical and Theoretical Chemistry
  • Colloid and Surface Chemistry

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