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Published September 2005 | public
Journal Article

A Nanoparticle-Based Model Delivery System To Guide the Rational Design of Gene Delivery to the Liver. 2. In Vitro and In Vivo Uptake Results

Abstract

In Part 1 of our work (1), four nanoparticles were synthesized specifically for the purpose of identifying design constraints to guide next generation gene delivery to the liver. The four nanoparticles are Gal-50 and Gal-140 (galactosylated 50 and 140 nm nanoparticles) and MeO-50 and MeO-140 (methoxy-terminated 50 and 140 nm nanoparticles). All four particles have the same surface charge, and Gal-50 and Gal-140 have the same surface galactose density (ca. 25−30 pmol/cm^2). Here, the hepatocyte uptake in vitro and hepatic distribution in vivo of these four nanoparticles is investigated. With freshly isolated hepatocytes, Gal-50 nanoparticles are taken up to a greater extent than are MeO-50, and both 50 nm beads are taken up to a much greater extent than either of the 140 nm nanoparticles. In mice, about 90% of the in vivo dose of Gal-140 nanoparticles is found within the liver 20 min after tail-vein injection. TEM and immunohistochemistry images confirm that Gal-140 nanoparticles are primarily internalized by Kupffer cells, though isolated examples of a few Gal-140 in hepatocytes are also observed. Gal-50 nanoparticles are overwhelmingly found in vesicles throughout the cytoplasm of hepatocytes, with only isolated examples of Kupffer cell uptake 20 min after tail vein injections in mice. Despite similar surface charge and ligand density, 50 nm nanoparticles are primarily found in hepatocytes while 140 nm nanoparticles are primarily observed in Kupffer cells. These results clearly show that slightly anionic, galactose-PEGylated nanoparticles with 25−30 pmol/cm^2 galactose should be about 50 nm in diameter to preferentially target hepatocytes while they should be about 140 nm in diameter to selectively target Kupffer cells.

Additional Information

© 2005 American Chemical Society. Received 14 April 2005. Published online 31 August 2005. Published in print 1 September 2005. We are grateful to Dr. Hide Tsukamoto for generously offering the services of his lab to isolate nonparenchymal liver cells and for helpful discussions. Dr. Rochelle Diamond went out of her way to assist with in vitro FACS experiments, and we thank George McNamara of the CHLARI Image Core for advice and assistance in acquiring images following immunhistochemical staining.

Additional details

Created:
August 19, 2023
Modified:
October 24, 2023