title: Combination of 3D tissue engineered scaffold and non-viral gene carrier enhance in vitro DNA expression of mesenchymal stem cells.
abstract: The objective of this study is to enhance the expression of a plasmid DNA for mesenchymal stem cells (MSC) by combination of 3-dimensional (3D) tissue engineered scaffolds and non-viral gene carrier. As a carrier of plasmid DNA, dextran-spermine cationic polysaccharide was prepared by means of reductive-amination between oxidized dextran and the natural oligoamine, spermine. As the MSC scaffold, collagen sponges reinforced by incorporation of poly(glycolic acid) (PGA) fibers were used. A complex of the cationized dextran and plasmid DNA of BMP-2 was impregnated into the scaffolds. MCS were seeded into each scaffold and cultured by a 3D culture method. When MSC were cultured in the PGA-reinforced sponge, the level of BMP-2 expression was significantly enhanced by the cationized dextran-plasmid DNA complex impregnated into the scaffold than by the cationized dextran-plasmid DNA complex in 2-dimensional (2D) (tissue culture plate) culture method. The alkaline phosphatase activity and osteocalcin content of transfected MSC cultured in the PGA-reinforced sponge were significantly higher compared with 2D culture method. We conclude that combination of cationized dextran plasmid DNA complex and 3D tissue engineered scaffold was promising to promote the in vitro gene expression for MSC.
authors.affiliation: International Center for Young Scientists (ICYS), National Institute for Materials Science (NIMS), Nano and Biomaterials Research Building, Namiki 1-1, Tsukuba, Ibaraki 305-0044, Japan.
authors.forename: Hossein
authors.initials: H
authors.lastname: Hosseinkhani
authors.forename: Tony
authors.initials: T
authors.lastname: Azzam
authors.forename: Hisatoshi
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Hiraoka
authors.forename: Hitoyata
authors.initials: H
authors.lastname: Shimokawa
authors.forename: Abraham J
authors.initials: AJ
authors.lastname: Domb
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tabata
mesh_terms: D000818:Animals; D004247:DNA / Q000378:metabolism*; D005822:Genetic Vectors; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D023822:Tissue Engineering; D014162:Transfection
chemical_list: D004247:DNA