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| Effect of Jet Nebulizer on Plasmid DNA Integrity and the Research of Protective Methods |
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Abstract Abstract Objective: To explore the effect of jet nebulizer on plasmid DNA (pDNA) integrity and the protective methods. Methods: In the experiment to determine the effect of nebulization time on pDNA integrity, 5ml pDNA solution was aerosolized. Aerosol samples were collected in the 1st min, 2nd min, 3rd min, 4th min, 5th min and the 10th min. In the experiment to determine the effect of aerosolized solution amount on pDNA integrity, 2 ml、4 ml、6 ml、8ml pDNA solution was aerosolized for 4 min respectively. Aerosol samples were collected in the last minute. In the experiment to determine the protective effectiveness of the two polyplex, 4 ml naked pDNA solution, 4ml PEI-DNA solution and 4 ml liposome-DNA solution was aerosolized for 10 min respectively. Aerosol samples were collected in the last minute. Degradation associated with the aerosolisation process was investigated with agarose gel electrophoresis. Results: As aerosolization time increased from 1min to 10min, the intact pDNA ratio decreased from (83.5±2.2)% to (37.1±2.8)%. As the pDNA solution amount added from 2 ml up to 8ml, the intact DNA ratio ascended from (32.1±3.5)% to (93.6±0.6)%. There was almost no damage to the PEI or cationic lipid protected pDNA solution. Conclusion: The damage to which jet nebulizer exerted on pDNA was related with time and dosage. PEI and cationic lipids provided plasmid DNA with excellent protection, which laid a good foundation for the application of jet nebulizer in future gene aerosol treatment.
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Received: 09 July 2010
Published: 19 October 2010
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| Fund: China Postdoctoral Science Foundation funded project |
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