title: Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer.
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.
authors.affiliation: Nanosystem Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan. a-oyane@aist.go.jp
authors.forename: Ayako
authors.initials: A
authors.lastname: Oyane
authors.forename: Yushin
authors.initials: Y
authors.lastname: Yazaki
authors.forename: Hiroko
authors.initials: H
authors.lastname: Araki
authors.forename: Yu
authors.initials: Y
authors.lastname: Sogo
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ito
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamazaki
authors.forename: Hideo
authors.initials: H
authors.lastname: Tsurushima
mesh_terms: D000818:Animals; D001031:Apatites / Q000737:chemistry*; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D008055:Lipids / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D013056:Spectrophotometry, Ultraviolet; D014961:X-Ray Diffraction
chemical_list: D001031:Apatites; D008055:Lipids; D004247:DNA