Mechanism of Cellular Uptake and Impact of Ferucarbotran on Macrophage Physiology

• Published in: PloS one Vol. 6; no. 9; p. e25524
• Main Authors: Yang, Chung-Yi, Tai, Ming-Fong, Lin, Chih-Peng, Lu, Chen-Wen, Wang, Jaw-Lin, Hsiao, Jong-Kai, Liu, Hon-Man
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.09.2011; Public Library of Science (PLoS)
• Subjects: Animals; Brain cancer; Cell Line; Cell proliferation; Cell Survival; Clathrin; Contrast agents; Contrast media; Contrast Media - pharmacology; Cytometry; Dextrans - pharmacokinetics; Dose-Response Relationship, Drug; Endocytosis; Ferric oxide; Flow cytometry; Flow Cytometry - methods; Iron; Iron content; Iron oxides; Labeling; Labelling; Macrophages; Macrophages - cytology; Macrophages - drug effects; Magnetic resonance; Magnetic resonance imaging; Magnetite Nanoparticles - chemistry; Materials Science; Medical imaging; Medical imaging equipment; Medicine; Membrane potential; Mice; Mitochondria; Monocytes - cytology; Nanoparticles; Nanoparticles - chemistry; NMR; Nuclear magnetic resonance; Oxygen; Particulates; Physiological aspects; Reactive Oxygen Species; Resonance; Rodents; Scattering; Scattering, Radiation; Spectrophotometers; Spectrophotometry - methods; Time Factors; United States > US; Taiwan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0025524

Superparamagnetic iron oxide (SPIO) nanoparticles are contrast agents used for magnetic resonance imaging. Ferucarbotran is a clinically approved SPIO-coated carboxydextran with a diameter of about 45-60 nm. We investigated the mechanism of cellular uptake of Ferucarbotran with a cell model using the murine macrophage cell line Raw 264.7. We observed a dose-dependent uptake of these SPIO particles by spectrophotometer analysis and also a dose-dependent increase in the granularity of the macrophages as determined by flow cytometry. There was a linear correlation between the side scattering mean value and iron content (P<0.001, R(2) = 0. 8048). For evaluation of the endocytotic pathway of these ingested SPIO particles, different inhibitors of the endocytotic pathways were employed. There was a significant decrease of side scattering counts in the cells and a less significant change in signal intensity based on magnetic resonance in the phenylarsine oxide-treated macrophages. After labeling with SPIO particles, the macrophages showed an increase in the production of reactive oxygen species at 2, 24, and 48 h; a decrease in mitochondrial membrane potential at 24 h; and an increase in cell proliferation at 24 h. We concluded that Ferucarbotran was internalized into macrophages via the clathrin-mediated pathway and can change the cellular behavior of these cells after labeling.