論文 - 詳細
RRC ID | 65105 |
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著者 | Sato T, Nakata M, Yang Z, Torizuka Y, Kishimoto S, Ishihara M. |
タイトル | In vitro and in vivo gene delivery using chitosan/hyaluronic acid nanoparticles: Influences of molecular mass of hyaluronic acid and lyophilization on transfection efficiency. |
ジャーナル | J Gene Med |
Abstract |
BACKGROUND:Lyophilization is an effective method for preserving nonviral gene vectors. To improve the stability and transgene expression of lyophilized plasmid DNA (pDNA) complexes, we coated the surfaces of pDNA/chitosan complexes with hyaluronic acid (HA) of varying molecular masses. The transgene expression of pDNA/chitosan/HA ternary complexes was characterized in vitro and in vivo. METHODS:pDNA complexes were lyophilized overnight and the resultant products with spongy, porous consistencies were stored at -30, 4 or 25°C for 2 weeks. Rehydrated complexes were characterized using gel retardation assays, aiming to confirm complex formation, measure particle size and evaluate zeta potential, as well as conduct luciferase gene reporter assays. The anti-tumor effects of pDNA ternary complexes were evaluated using suicide gene (pTK) coding thymidine kinase in Huh7-implanted mice. RESULTS:Transfection efficiencies of pDNA/chitosan/HA ternary complexes were dependent on the average molecular masses of HA. The coating of pDNA/chitosan complexes with HA maintained the cellular transfection efficiencies of lyophilized pDNA ternary complexes. Furthermore, intratumoral injection of lyophilized, rehydrated pDNA ternary complexes into tumor-bearing mice showed a significant suppression of tumor growth. CONCLUSIONS:The coating of pDNA/chitosan complexes with high-molecular-weight HA augmented the stability and cellular transfection ability of the complexes after lyophilization-rehydration. |
巻・号 | 19(8) |
公開日 | 2017-8-1 |
DOI | 10.1002/jgm.2968 |
PMID | 28667693 |
MeSH | Animals Chitosan / therapeutic use* DNA / genetics Electrophoretic Mobility Shift Assay Freeze Drying Gene Transfer Techniques Genes, Reporter Genetic Therapy / methods* Genetic Vectors Humans Hyaluronic Acid / therapeutic use* Luciferases / genetics Mice Nanoparticles Particle Size Plasmids Thymidine Kinase / genetics Transfection |
IF | 3.258 |
リソース情報 | |
ヒト・動物細胞 | HuH-7(RCB1366) |