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 journalArticle

    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

    Fingerprint

    Gene transfer
    Apatites
    Apatite
    apatites
    genes
    Lipids
    lipids
    DNA
    deoxyribonucleic acid
    Immobilized Nucleic Acids
    composite materials
    Composite materials
    Chemical analysis
    Genes
    Coprecipitation
    CHO Cells
    Bioelectric potentials
    Tissue Engineering
    calcium phosphates
    Tissue engineering

    Keywords

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

    ASJC Scopus subject areas

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

    Cite this

    Improved gene transfer efficiency of a DNA-lipid-apatite composite layer by controlling the layer molecular composition. / Yazaki, Yushin; Oyane, Ayako; Tsurushima, Hideo; Araki, Hiroko; Sogo, Yu; Ito, Atsuo; Yamazaki, Atsushi.

    In: Colloids and Surfaces B: Biointerfaces, Vol. 122, 01.10.2014, p. 465-471.

    Research output: Contribution to journalArticle

    Yazaki, Yushin ; Oyane, Ayako ; Tsurushima, Hideo ; Araki, Hiroko ; Sogo, Yu ; Ito, Atsuo ; Yamazaki, Atsushi. / Improved gene transfer efficiency of a DNA-lipid-apatite composite layer by controlling the layer molecular composition. In: Colloids and Surfaces B: Biointerfaces. 2014 ; Vol. 122. pp. 465-471.
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