Ethanol enhanced in vivo gene delivery with non-ionic polymeric micelles inhalation.
Modifications of both carriers and host barriers have been investigated for efficient inhalation gene delivery to lung. Here we used a biocompatible, non-ionic poly(ethyleneoxide)-poly(propyleneoxide)-poly(ethyleneoxide) (PEO-PPO-PEO) polymeric micelles (PM) as a carrier and combined it with ethanol to enhance membrane penetration of delivered DNA. The inhalation delivery with six 100 microg doses of pCMV-Lac Z with PM co-formulated with 10%-40% ethanol to nude mice in 2 days at 8 h interval was performed. The beta-galatosidase (beta-Gal) activity was assessed using chlorophenol red-beta-d galactopyranoside (CPRG) and X-gal staining for quantitative and qualitative analysis in tissues. The results showed that beta-Gal activity was significantly increased by 38% in lung around bronchioles when inhalation with PM and 10% ethanol was given. The 10% ethanol also increased the intracellular apparent permeability by 42% in stomach and by 141% in intestine at 48 h after the first dosage of delivery. Also delivery of DNA encoding a functional human cystic fibrosis transmembrane protein (CFTR) using the same inhalation delivery method co-formulated with 10% ethanol, an increased expression of CFTR in lung was detected by immunostaining. We concluded that 10% ethanol co-formulated with the PM system could enhance inhaled gene delivery to airway and gastrointestinal (GI) tract.
Chao
Yen-Chin
YC
College of Pharmacy, Taipei Medical University, 250 Wu Hsing Street, Taipei 110, Taiwan.
Chang
Shwu-Fen
SF
Lu
Shao-Chun
SC
Hwang
Tzyh-Chang
TC
Hsieh
Wei-Hsien
WH
Liaw
Jiahorng
J
D000280:Administration, Inhalation; D000818:Animals; D004247:DNA; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000737:chemistry*; D018014:Gene Transfer Techniques; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008823:Micelles; D018625:Microscopy, Atomic Force; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols / Q000008:administration & dosage* / Q000737:chemistry; D011108:Polymers / Q000008:administration & dosage / Q000737:chemistry; D011409:Propylene Glycols / Q000008:administration & dosage* / Q000737:chemistry; D013047:Specific Pathogen-Free Organisms; D001616:beta-Galactosidase / Q000378:metabolism
D008823:Micelles; C116176:PEO-PPO-PEO; D011108:Polymers; D011409:Propylene Glycols; D000431:Ethanol; D011092:Polyethylene Glycols; D004247:DNA; D001616:beta-Galactosidase