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Chitosan-DNA nanoparticles delivered by intrabiliary infusion enhance liver-targeted gene delivery.

Dai H, Jiang X, Tan GC, Chen Y, Torbenson M, Leong KW, Mao HQ - Int J Nanomedicine (2006)

Bottom Line: Chitosan-DNA nanoparticles administrated through the portal vein or tail vein did not produce detectable luciferase expression.Luciferase expression in the kidney, lung, spleen, and heart was negligible compared with that in the liver.These results suggest that RII is a promising routine to achieve liver-targeted gene delivery by non-viral nanoparticles; and both gene carrier characteristics and mode of administration significantly influence gene delivery efficiency.

View Article: PubMed Central - PubMed

Affiliation: Tissue and Therapeutic Engineering Lab, Division of Johns Hopkins in Singapore, Singapore.

ABSTRACT
The goal of this study was to examine the efficacy of liver-targeted gene delivery by chitosan-DNA nanoparticles through retrograde intrabiliary infusion (RII). The transfection efficiency of chitosan-DNA nanoparticles, as compared with PEI-DNA nanoparticles or naked DNA, was evaluated in Wistar rats by infusion into the common bile duct, portal vein, or tail vein. Chitosan-DNA nanoparticles administrated through the portal vein or tail vein did not produce detectable luciferase expression. In contrast, rats that received chitosan-DNA nanoparticles showed more than 500 times higher luciferase expression in the liver 3 days after RII; and transgene expression levels decreased gradually over 14 days. Luciferase expression in the kidney, lung, spleen, and heart was negligible compared with that in the liver. RII of chitosan-DNA nanoparticles did not yield significant toxicity and damage to the liver and biliary tree as evidenced by liver function analysis and histopathological examination. Luciferase expression by RII of PEI-DNA nanoparticles was 17-fold lower than that of chitosan-DNA nanoparticles on day 3, but it increased slightly over time. These results suggest that RII is a promising routine to achieve liver-targeted gene delivery by non-viral nanoparticles; and both gene carrier characteristics and mode of administration significantly influence gene delivery efficiency.

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Related in: MedlinePlus

Confocal fluorescence images indicating DNA distribution in the right lobe of the liver that received naked DNA and nanoparticles via RII. Liver samples were collected 4 hours after intrabiliary infusion of chitosan-DNA or PEI-DNA nanoparticles containing 200 μg of Cy5 labeled DNA (pGeneGrip) or 200 μg of naked Cy5-DNA. (a–c): distribution of DNA (red); (d–h): co-localization of DNA (red) and KCs (green). KCs were stained with FITC-labeled mouse anti-rat macrophage F-6-J mAb. (i–k): co-localization of DNA (red) and endothelial cells (ECs, green). Endothelial cells were identified by immunostaining with FITC-labeled mouse anti-rat RECA-1 mAb.Abbreviation: PEI, polyethylenimine; EC, endothelial cell; KC, Kupffer cell.
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f7-ijn-1-507: Confocal fluorescence images indicating DNA distribution in the right lobe of the liver that received naked DNA and nanoparticles via RII. Liver samples were collected 4 hours after intrabiliary infusion of chitosan-DNA or PEI-DNA nanoparticles containing 200 μg of Cy5 labeled DNA (pGeneGrip) or 200 μg of naked Cy5-DNA. (a–c): distribution of DNA (red); (d–h): co-localization of DNA (red) and KCs (green). KCs were stained with FITC-labeled mouse anti-rat macrophage F-6-J mAb. (i–k): co-localization of DNA (red) and endothelial cells (ECs, green). Endothelial cells were identified by immunostaining with FITC-labeled mouse anti-rat RECA-1 mAb.Abbreviation: PEI, polyethylenimine; EC, endothelial cell; KC, Kupffer cell.

Mentions: To characterize the distribution of nanoparticles and naked DNA following RII, plasmid DNA was labeled with Cy5 using pGeneGrip technology before nanoparticle preparation. Tissues from major organs were harvested 4 hours after infusion, and cryo-sectioned for the examination of Cy5-labelled plasmid. For animals that received nanoparticles, the reporter gene was detected throughout the liver, although the pattern was rather heterogeneous; occasionally clusters of Cy5 fluorescence were found in close proximity to vessel structure or portal triads (Figures 7a, b). Only a low level of reporter gene was found in naked DNA-transfected rats, highlighting the importance of gene carriers. Comparing the two nanoparticle groups, more fluorescence was observed for the PEI-DNA nanoparticle group than the chitosan-DNA nanoparticle group. The relative levels of reporter gene observed in the tissue, however, seemed to contradict the relative levels of transgene expression in the liver: chitosan-DNA nanoparticles showed a lower level of presence in the liver, but yielded higher level of luciferase expression, in contrast to PEI-DNA nanoparticles. This may be due to the fact that chitosan is degradable, therefore resulting in higher level of DNA release following cell uptake.


Chitosan-DNA nanoparticles delivered by intrabiliary infusion enhance liver-targeted gene delivery.

Dai H, Jiang X, Tan GC, Chen Y, Torbenson M, Leong KW, Mao HQ - Int J Nanomedicine (2006)

Confocal fluorescence images indicating DNA distribution in the right lobe of the liver that received naked DNA and nanoparticles via RII. Liver samples were collected 4 hours after intrabiliary infusion of chitosan-DNA or PEI-DNA nanoparticles containing 200 μg of Cy5 labeled DNA (pGeneGrip) or 200 μg of naked Cy5-DNA. (a–c): distribution of DNA (red); (d–h): co-localization of DNA (red) and KCs (green). KCs were stained with FITC-labeled mouse anti-rat macrophage F-6-J mAb. (i–k): co-localization of DNA (red) and endothelial cells (ECs, green). Endothelial cells were identified by immunostaining with FITC-labeled mouse anti-rat RECA-1 mAb.Abbreviation: PEI, polyethylenimine; EC, endothelial cell; KC, Kupffer cell.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC1828073&req=5

f7-ijn-1-507: Confocal fluorescence images indicating DNA distribution in the right lobe of the liver that received naked DNA and nanoparticles via RII. Liver samples were collected 4 hours after intrabiliary infusion of chitosan-DNA or PEI-DNA nanoparticles containing 200 μg of Cy5 labeled DNA (pGeneGrip) or 200 μg of naked Cy5-DNA. (a–c): distribution of DNA (red); (d–h): co-localization of DNA (red) and KCs (green). KCs were stained with FITC-labeled mouse anti-rat macrophage F-6-J mAb. (i–k): co-localization of DNA (red) and endothelial cells (ECs, green). Endothelial cells were identified by immunostaining with FITC-labeled mouse anti-rat RECA-1 mAb.Abbreviation: PEI, polyethylenimine; EC, endothelial cell; KC, Kupffer cell.
Mentions: To characterize the distribution of nanoparticles and naked DNA following RII, plasmid DNA was labeled with Cy5 using pGeneGrip technology before nanoparticle preparation. Tissues from major organs were harvested 4 hours after infusion, and cryo-sectioned for the examination of Cy5-labelled plasmid. For animals that received nanoparticles, the reporter gene was detected throughout the liver, although the pattern was rather heterogeneous; occasionally clusters of Cy5 fluorescence were found in close proximity to vessel structure or portal triads (Figures 7a, b). Only a low level of reporter gene was found in naked DNA-transfected rats, highlighting the importance of gene carriers. Comparing the two nanoparticle groups, more fluorescence was observed for the PEI-DNA nanoparticle group than the chitosan-DNA nanoparticle group. The relative levels of reporter gene observed in the tissue, however, seemed to contradict the relative levels of transgene expression in the liver: chitosan-DNA nanoparticles showed a lower level of presence in the liver, but yielded higher level of luciferase expression, in contrast to PEI-DNA nanoparticles. This may be due to the fact that chitosan is degradable, therefore resulting in higher level of DNA release following cell uptake.

Bottom Line: Chitosan-DNA nanoparticles administrated through the portal vein or tail vein did not produce detectable luciferase expression.Luciferase expression in the kidney, lung, spleen, and heart was negligible compared with that in the liver.These results suggest that RII is a promising routine to achieve liver-targeted gene delivery by non-viral nanoparticles; and both gene carrier characteristics and mode of administration significantly influence gene delivery efficiency.

View Article: PubMed Central - PubMed

Affiliation: Tissue and Therapeutic Engineering Lab, Division of Johns Hopkins in Singapore, Singapore.

ABSTRACT
The goal of this study was to examine the efficacy of liver-targeted gene delivery by chitosan-DNA nanoparticles through retrograde intrabiliary infusion (RII). The transfection efficiency of chitosan-DNA nanoparticles, as compared with PEI-DNA nanoparticles or naked DNA, was evaluated in Wistar rats by infusion into the common bile duct, portal vein, or tail vein. Chitosan-DNA nanoparticles administrated through the portal vein or tail vein did not produce detectable luciferase expression. In contrast, rats that received chitosan-DNA nanoparticles showed more than 500 times higher luciferase expression in the liver 3 days after RII; and transgene expression levels decreased gradually over 14 days. Luciferase expression in the kidney, lung, spleen, and heart was negligible compared with that in the liver. RII of chitosan-DNA nanoparticles did not yield significant toxicity and damage to the liver and biliary tree as evidenced by liver function analysis and histopathological examination. Luciferase expression by RII of PEI-DNA nanoparticles was 17-fold lower than that of chitosan-DNA nanoparticles on day 3, but it increased slightly over time. These results suggest that RII is a promising routine to achieve liver-targeted gene delivery by non-viral nanoparticles; and both gene carrier characteristics and mode of administration significantly influence gene delivery efficiency.

Show MeSH
Related in: MedlinePlus