Kupffer cells are central in the removal of nanoparticles from the organism

Abstract

Background: The study aims at revealing the fate of nanoparticles administered intravenously and intraperitoneally to adult female mice, some of which were pregnant. Gold nanoparticles were chosen as a model because these particles have been found to be chemically inert and at the same time are easily traced by autometallography (AMG) at both ultrastructural and light microscopic levels.

Results: Gold nanoparticles were injected intravenously (IV) or intraperitoneally (IP) and traced after 1, 4 or 24 hours. For IV injections 2 and 40 nm particles were used; for IP injections 40 nm particles only. The injected nanoparticles were found in macrophages only, and at moderate exposure primarily in the Kupffer cells in the liver. IV injections resulted in a rapid accumulation/clustering of nanoparticles in these liver macrophages, while the uptake in spleen macrophages was moderate. IP injections were followed by a delayed uptake in the liver and included a moderate uptake in macrophages located in mesenteric lymph nodes, spleen and small intestine. Ultrastructurally, the AMG silver enhanced nanocrystals were found in lysosome-like organelles of the Kupffer cells and other macrophages wherever located.Accumulations of gold nanoparticles were not found in any other organs analysed, i.e. kidneys, brain, lungs, adrenals, ovaries, placenta, and fetal liver, and the control animals were all void of AMG staining.

Conclusion: Our results suggest that: (1) inert gold nanoparticles do not penetrate cell membranes by non-endocytotic mechanisms, but are rather taken up by endocytosis; (2) gold nanoparticles, independent of size, are taken up primarily by Kupffer cells in the liver and secondarily by macrophages in other places; (3) gold nanoparticles do not seem to penetrate the placenta barrier; (4) the blood-brain barrier seems to protect the central nervous system from gold nanoparticles; (5) 2 nanometer gold particles seem to be removed not only by endocytosis by macrophages, and we hypothesize that part of these tiny nanoparticles are released into the urine as a result of simple filtration in the renal glomeruli.

Figure 1

Micrographs demonstrating AMG silver enhanced clustered gold nanoparticles in mouse after in vivo exposure. (a) 40 nm gold particles clustered in Kupffer cells of the liver from a pregnant mouse. (b) Section from a fetal liver taken from an embryo of the same animal. Note that the fetal tissue is completely void of staining. The pregnant animal was intravenously injected 40 nm gold particles and allowed to survive for 24 hours. Both sections were 3 micron Epon sections counterstained with toluidine blue. (c) Section from the liver of a pregnant mouse, which served as a control and was exposed to saline. The section is completely void of AMG staining; (d) Micrograph of nanogold particles in a spleen macrophage. The animal was treated intravenously with 40 nm gold particles 24 hours before being sacrificed. 3 micron Epon section counterstained with Toluidine blue. (e) Enhanced gold nanoparticles in macrophages of a mesenterial lymph node. The animal was injected 40 nm gold nanoparticles intraperitoneally and allowed to survive for 4 hours, 30 μm thick cryo section, counterstained with toluidine blue. (f) Micrograph of a mesenterial lymph node of a mouse which was exposed to saline intraperitoneally and served as control. Scalebars = 20 μm.

Figure 2

Electron micrographs showing AMG enhanced clustered gold nanoparticles in the lysosomes of a Kupffer cell (a) and a spleen macrophage (b). The animal was exposed to 40 nm gold nanoparticles intravenously and allowed to survive for 24 hours. Scalebars = 2 μm.