title: DNA-lipid-apatite composite layers enhance gene expression of mesenchymal stem cells.
abstract: DNA-apatite composite layer (D-Ap layer) and DNA-lipid-apatite composite layer (DLp-Ap layer) were prepared on ceramic hydroxyapatite disk and scaffold using supersaturated calcium phosphate solutions supplemented with 0.5-5 μg/mL plasmid and 0-10 μL/mL lipid transfection reagent FuGENE®. Both in vitro and in vivo studies were carried out using mesenchymal stem cells (MSCs) and two kinds of gene (luciferase and bone morphogenetic protein (BMP)-2) for demonstrating potential application of the gene transfer system using the D-Ap and DLp-Ap layers in bone tissue engineering. In the in vitro study using luciferase gene, the DLp-Ap layers showed 1-2 orders of magnitudes higher gene transfer efficiency to MSCs than the D-Ap layer. In the in vivo study using BMP-2 gene, DLp-Ap layer slightly increased BMP-2 protein concentration than D-Ap layer, thereby enhancing their osteogenic differentiation than D-Ap layer. The present gene transfer system using the DLp-Ap layers, with the advantages of good biocompatibility, bone-bonding ability, and efficacy in in vitro and in vivo gene transfer to MSCs, would be useful in bone tissue engineering.
authors.affiliation: Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. xp-wang@aist.go.jp
authors.forename: Xiupeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ito
authors.affiliation: Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sogo
authors.affiliation: Human Technology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Hirose
authors.affiliation: Nanosystem Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Oyane
authors.affiliation: Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Tsurushima
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001031:Apatites / Q000494:pharmacology*; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D017886:Durapatite / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D008055:Lipids / Q000494:pharmacology*; D008156:Luciferases / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D015675:Osteocalcin / Q000378:metabolism; D011916:Rats, Inbred F344; D012867:Skin / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry; D014162:Transfection; D014961:X-Ray Diffraction
chemical_list: D001031:Apatites; D055396:Bone Morphogenetic Protein 2; D008055:Lipids; D015675:Osteocalcin; D004247:DNA; D017886:Durapatite; D008156:Luciferases; D000469:Alkaline Phosphatase