Pulmonary gene delivery by chitosan-pDNA complex powder prepared by a supercritical carbon dioxide process.
Chitosan-plasmid DNA (pDNA) complex powders as a pulmonary gene delivery system were prepared with a supercritical carbon dioxide (CO(2)) process and their in vivo activity was evaluated. The powders with mannitol as a carrier were prepared by dispersing aqueous solutions of a luciferase expression plasmid driven by the cytomegalovirus promoter (pCMV-Luc) with or without chitosan as a cationic vector in a supercritical CO(2)/ethanol admixture. The supercritical CO(2) process with a V-shaped nozzle successfully produced chitosan-pDNA powders. The addition of chitosan suppressed the degradation of pCMV-Luc during the supercritical CO(2) process and increased the yield of powders. The luciferase activity in mouse lung was evaluated after pulmonary administration of the powders or pCMV-Luc solutions. The chitosan-pDNA powders increased the luciferase activity in mouse lung compared with pCMV-Luc powders without chitosan or pCMV-Luc solutions with or without chitosan. The chitosan-pDNA powder with an N/P ratio = 5 increased the luciferase activity to 2700% of that of the pCMV-Luc solution. These results suggest that gene powder with chitosan is a useful pulmonary gene delivery system.
Okamoto
Hirokazu
H
Faculty of Pharmacy, Meijo University, 150 Yagotoyama Tempaku-ku, Nagoya 468-8503, Japan. okamotoh@ccmfs.meijo-u.ac.jp
Nishida
Seiko
S
Todo
Hiroaki
H
Sakakura
Yuki
Y
Iida
Kotaro
K
Danjo
Kazumi
K
D000818:Animals; D002245:Carbon Dioxide / Q000737:chemistry*; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry*; D048271:Chitosan; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D005079:Excipients; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics; D008168:Lung / Q000502:physiology*; D051379:Mice; D008813:Mice, Inbred ICR; D008855:Microscopy, Electron, Scanning; D010316:Particle Size; D010957:Plasmids / Q000235:genetics*; D011208:Powders; D011401:Promoter Regions, Genetic / Q000235:genetics; D014162:Transfection
D005079:Excipients; D011208:Powders; D002686:Chitin; D002245:Carbon Dioxide; D004247:DNA; D048271:Chitosan; D008156:Luciferases