Use of perfluorocarbon (fluorinert) to enhance reporter gene expression following intratracheal instillation into the lungs of Balb/c mice: Implications for nebulized delivery of plasmids
✍ Scribed by Aditya Das; Ralph Niven
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 923 KB
- Volume
- 90
- Category
- Article
- ISSN
- 0022-3549
- DOI
- 10.1002/jps.1086
No coin nor oath required. For personal study only.
✦ Synopsis
Per¯uorocarbons combine high respiratory gas dissolving capabilities with extreme chemical and biological inertness and therefore offer an attractive option as an excipient in the area of pulmonary therapeutics. Per¯uorocarbons have also been shown to `¯oat' mucus, because of their high densities (1.9±2.5 g/mL), which may hold potential in gene delivery for cystic ®brosis patients, in terms of enhancing penetration through highly viscous mucus and thereby providing access to target epithelial cells to correct the gene defect. Additionally, their low surface tension allows for better dispersion. A commonly available per¯urocarbon, heptacosa¯uorotributylamine (Fluorinert), was used to deliver either plasmid DNA (pDNA) alone or cationic-lipid-complexed plasmid DNA to the lungs of Balb/c mice by direct intratracheal instillation. The complexes consisted of supercoiled (SC) plasmid DNA (4.7 Kb, 0.625 mg/mL) and lipid (ethyldimyristoyl phosphatidylcholine [EDMPC]/cholesterol [1:1 mole ratio], with pDNA (3:1 mg pDNA/mM EDMPC in 20 mM Tris-HCl pH 8.0) expressing chloramphenicol acetyl transferase (CAT) or b-galactosidase (b-Gal). pDNA alone was supplemented with 14% w/v Fluorinert. Cationic lipid/pDNA complexes were supplemented with 3, 8, and 14% w/v Fluorinert. Results showed that the CAT expression from pDNA alone was enhanced 24 Â using 14% w/v Fluorinert, whereas that from the cationic-lipidformulated pDNA was enhanced 7 Â using 14% w/v Fluorinert. Immunohistochemistry showed that b-Gal expression was primarily from epithelial cells and not from F4/80 or MAC3 antigen-stained cells (predominantly macrophages), indicating ef®cient delivery.