TY - JOUR
T1 - Improved gene transfer efficiency of a DNA-lipid-apatite composite layer by controlling the layer molecular composition
AU - Yazaki, Yushin
AU - Oyane, Ayako
AU - Tsurushima, Hideo
AU - Araki, Hiroko
AU - Sogo, Yu
AU - Ito, Atsuo
AU - Yamazaki, Atsushi
N1 - Funding Information:
This work was supported in part by a JSPS KAKENHI: Grant-in-Aid for young scientist (B) (22700499) and in part by a Research Grant from the Foundation, Oil & Fat Industry Kaikan , Japan. We acknowledge Katayama Chemical Industries, Co., Ltd., Japan for supplying the Lps (PD and mPD). A part of this work was conducted at the Nano-Processing Facility, which is supported by the IBEC Innovation Platform, National Institute of Advanced Industrial Science and Technology (AIST) , Japan.
Publisher Copyright:
© 2014 Elsevier B.V.
PY - 2014/10/1
Y1 - 2014/10/1
N2 - 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.
AB - 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.
KW - Apatite
KW - Composite
KW - DNA
KW - Gene transfer
KW - Lipid
KW - Lipofection
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U2 - 10.1016/j.colsurfb.2014.07.001
DO - 10.1016/j.colsurfb.2014.07.001
M3 - Article
C2 - 25096721
AN - SCOPUS:84910091919
SN - 0927-7765
VL - 122
SP - 465
EP - 471
JO - Colloids and Surfaces B: Biointerfaces
JF - Colloids and Surfaces B: Biointerfaces
ER -