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Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer

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Abstract

A surface-mediated gene transfer system using biocompatible apatite-based composite layers has great potential for tissue engineering. Among the apatite-based composite layers developed to date, we focused on a DNA-lipid-apatite composite layer (DLp-Ap layer), which has the advantage of relatively high efficiency as a non-viral system. In this study, various lipid transfection reagents, including a newly developed reagent, polyamidoamine dendron-bearing lipid (PD), were employed to prepare the DLp-Ap layer, and the preparation condition was optimized in terms of efficiency of gene transfer to epithelial-like CHO-K1 cells in the presence of serum. The optimized DLp-Ap layer derived from PD had the highest gene transfer efficiency among all the apatite-based composite layers prepared in this study. In addition, the optimized DLp-Ap layer demonstrated higher gene transfer efficiency in the presence of serum than the conventional particle-mediated systems using commercially available lipid transfection reagents. It was also shown that the optimized DLp-Ap layer mediated the area-specific gene transfer on its surface, i.e., DNA was preferentially transferred to the cells adhering to the surface of the layer. The present gene transfer system using the PD-derived DLp-Ap layer, with the advantages of high efficiency in the presence of serum and area-specificity, would be useful in tissue engineering.

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Acknowledgments

This work was supported by Grant-in-Aid for young scientist (B) (22700499) from the Ministry of Education, Culture, Sport, Science and Technology of Japan. We acknowledge Katayama Chemical Industries, Co., Ltd., Japan for supplying PD. A part of this work was conducted at the Nano-Processing Facility, supported by IBEC Innovation Platform, National Institute of Advanced Industrial Science and Technology (AIST), Japan.

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Authors and Affiliations

  1. Nanosystem Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8562, Japan

    Ayako Oyane, Hiroko Araki & Hideo Tsurushima

  2. Department of Resources and Environmental Engineering, Waseda University, 3-4-1 Ohkubo, Shinjuku, Tokyo, 169-8555, Japan

    Yushin Yazaki & Atsushi Yamazaki

  3. Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Yu Sogo & Atsuo Ito

  4. Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki, 305-8575, Japan

    Hideo Tsurushima

Authors
  1. Ayako Oyane
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  2. Yushin Yazaki
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  3. Hiroko Araki
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  4. Yu Sogo
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  5. Atsuo Ito
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  6. Atsushi Yamazaki
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  7. Hideo Tsurushima
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Correspondence to Ayako Oyane or Hideo Tsurushima.

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Oyane, A., Yazaki, Y., Araki, H. et al. Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer. J Mater Sci: Mater Med 23, 1011–1019 (2012). https://doi.org/10.1007/s10856-012-4581-y

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  • DOI: https://doi.org/10.1007/s10856-012-4581-y

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