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Evaluation of uptake and transport of cationic and anionic ultrasmall iron oxide nanoparticles by human colon cells.

Kenzaoui BH, Vilà MR, Miquel JM, Cengelli F, Juillerat-Jeanneret L - Int J Nanomedicine (2012)

Bottom Line: The intracellular localization of aminoPVA USPIO NPs was confirmed in HT-29 cells by transmission electron microscopy that detected the iron oxide core.AminoPVA USPIO NPs invaded three-dimensional spheroids of both HT-29 and Caco-2 cells, whereas oleic acid-coated USPIO NPs could only invade Caco-2 spheroids.Neither cationic aminoPVA USPIO NPs nor anionic oleic acid-coated USPIO NPs were transported at detectable levels across the tight CacoReady™ intestinal barrier model or the more permeable mucus-secreting CacoGoblet™ model.

View Article: PubMed Central - PubMed

Affiliation: Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.

ABSTRACT
Nanoparticles (NPs) are in clinical use or under development for therapeutic imaging and drug delivery. However, relatively little information exists concerning the uptake and transport of NPs across human colon cell layers, or their potential to invade three-dimensional models of human colon cells that better mimic the tissue structures of normal and tumoral colon. In order to gain such information, the interactions of biocompatible ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) (iron oxide core 9-10 nm) coated with either cationic polyvinylamine (aminoPVA) or anionic oleic acid with human HT-29 and Caco-2 colon cells was determined. The uptake of the cationic USPIO NPs was much higher than the uptake of the anionic USPIO NPs. The intracellular localization of aminoPVA USPIO NPs was confirmed in HT-29 cells by transmission electron microscopy that detected the iron oxide core. AminoPVA USPIO NPs invaded three-dimensional spheroids of both HT-29 and Caco-2 cells, whereas oleic acid-coated USPIO NPs could only invade Caco-2 spheroids. Neither cationic aminoPVA USPIO NPs nor anionic oleic acid-coated USPIO NPs were transported at detectable levels across the tight CacoReady™ intestinal barrier model or the more permeable mucus-secreting CacoGoblet™ model.

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Time-dependent uptake of oleic acid-coated ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) by Caco-2 and HT-29 cells. (A) Caco-2 cells were grown to 75% confluence, then were exposed for 6 or 24 hours in complete culture medium to 20 μg/mL oleic acid-coated USPIO NPs. Then cells were stained with Prussian blue and Nuclear Red histological stains (iron blue, nucleus red, cytoplasm pink). (B) Caco-2 and HT-29 cells were grown to 90% confluence, then they were exposed for 6 hours (grey line) or 24 hours (black line) in complete culture medium to increasing concentrations of oleic acid-coated USPIO NPs and the cell-associated iron content of the cell layer was quantified using the Prussian blue reaction.
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f2-ijn-7-1275: Time-dependent uptake of oleic acid-coated ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) by Caco-2 and HT-29 cells. (A) Caco-2 cells were grown to 75% confluence, then were exposed for 6 or 24 hours in complete culture medium to 20 μg/mL oleic acid-coated USPIO NPs. Then cells were stained with Prussian blue and Nuclear Red histological stains (iron blue, nucleus red, cytoplasm pink). (B) Caco-2 and HT-29 cells were grown to 90% confluence, then they were exposed for 6 hours (grey line) or 24 hours (black line) in complete culture medium to increasing concentrations of oleic acid-coated USPIO NPs and the cell-associated iron content of the cell layer was quantified using the Prussian blue reaction.

Mentions: Previous research has shown that positive charges at the surface of aminoPVA-USPIO NPs are necessary for their uptake by cells, including human cells, since aminoPVA- coated (but not carboxylate-PVA-coated or PVA-coated) USPIO NPs were internalized by the cells.14,16 Therefore, the association of these previously described cationic aminoPVA USPIO NPs (Figure 1) with HT-29 and Caco-2 human colon cells was compared to their association with anionic oleic acid-coated USPIO NPs (Figure 2), as determined through the evaluation of cell-associated iron by histological or quantitative Prussian blue reaction. In both cell lines, the association of aminoPVA USPIO NPs dose-dependently increased with time (Figure 1), while anionic USPIO NPs were only poorly associated with colon cells and this association did not increase with longer exposure time (Figure 2). For these USPIO NPs, the iron content in the cell layers was below the detection limit of the Prussian blue method. Therefore, cationic USPIO NPs associated with human colon cells to a larger extent than anionic USPIO NPs. Similar results were obtained in both cell lines (data of the histological Prussian blue reaction are not shown for HT-29 cells).


Evaluation of uptake and transport of cationic and anionic ultrasmall iron oxide nanoparticles by human colon cells.

Kenzaoui BH, Vilà MR, Miquel JM, Cengelli F, Juillerat-Jeanneret L - Int J Nanomedicine (2012)

Time-dependent uptake of oleic acid-coated ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) by Caco-2 and HT-29 cells. (A) Caco-2 cells were grown to 75% confluence, then were exposed for 6 or 24 hours in complete culture medium to 20 μg/mL oleic acid-coated USPIO NPs. Then cells were stained with Prussian blue and Nuclear Red histological stains (iron blue, nucleus red, cytoplasm pink). (B) Caco-2 and HT-29 cells were grown to 90% confluence, then they were exposed for 6 hours (grey line) or 24 hours (black line) in complete culture medium to increasing concentrations of oleic acid-coated USPIO NPs and the cell-associated iron content of the cell layer was quantified using the Prussian blue reaction.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3298390&req=5

f2-ijn-7-1275: Time-dependent uptake of oleic acid-coated ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) by Caco-2 and HT-29 cells. (A) Caco-2 cells were grown to 75% confluence, then were exposed for 6 or 24 hours in complete culture medium to 20 μg/mL oleic acid-coated USPIO NPs. Then cells were stained with Prussian blue and Nuclear Red histological stains (iron blue, nucleus red, cytoplasm pink). (B) Caco-2 and HT-29 cells were grown to 90% confluence, then they were exposed for 6 hours (grey line) or 24 hours (black line) in complete culture medium to increasing concentrations of oleic acid-coated USPIO NPs and the cell-associated iron content of the cell layer was quantified using the Prussian blue reaction.
Mentions: Previous research has shown that positive charges at the surface of aminoPVA-USPIO NPs are necessary for their uptake by cells, including human cells, since aminoPVA- coated (but not carboxylate-PVA-coated or PVA-coated) USPIO NPs were internalized by the cells.14,16 Therefore, the association of these previously described cationic aminoPVA USPIO NPs (Figure 1) with HT-29 and Caco-2 human colon cells was compared to their association with anionic oleic acid-coated USPIO NPs (Figure 2), as determined through the evaluation of cell-associated iron by histological or quantitative Prussian blue reaction. In both cell lines, the association of aminoPVA USPIO NPs dose-dependently increased with time (Figure 1), while anionic USPIO NPs were only poorly associated with colon cells and this association did not increase with longer exposure time (Figure 2). For these USPIO NPs, the iron content in the cell layers was below the detection limit of the Prussian blue method. Therefore, cationic USPIO NPs associated with human colon cells to a larger extent than anionic USPIO NPs. Similar results were obtained in both cell lines (data of the histological Prussian blue reaction are not shown for HT-29 cells).

Bottom Line: The intracellular localization of aminoPVA USPIO NPs was confirmed in HT-29 cells by transmission electron microscopy that detected the iron oxide core.AminoPVA USPIO NPs invaded three-dimensional spheroids of both HT-29 and Caco-2 cells, whereas oleic acid-coated USPIO NPs could only invade Caco-2 spheroids.Neither cationic aminoPVA USPIO NPs nor anionic oleic acid-coated USPIO NPs were transported at detectable levels across the tight CacoReady™ intestinal barrier model or the more permeable mucus-secreting CacoGoblet™ model.

View Article: PubMed Central - PubMed

Affiliation: Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.

ABSTRACT
Nanoparticles (NPs) are in clinical use or under development for therapeutic imaging and drug delivery. However, relatively little information exists concerning the uptake and transport of NPs across human colon cell layers, or their potential to invade three-dimensional models of human colon cells that better mimic the tissue structures of normal and tumoral colon. In order to gain such information, the interactions of biocompatible ultrasmall superparamagnetic iron oxide nanoparticles (USPIO NPs) (iron oxide core 9-10 nm) coated with either cationic polyvinylamine (aminoPVA) or anionic oleic acid with human HT-29 and Caco-2 colon cells was determined. The uptake of the cationic USPIO NPs was much higher than the uptake of the anionic USPIO NPs. The intracellular localization of aminoPVA USPIO NPs was confirmed in HT-29 cells by transmission electron microscopy that detected the iron oxide core. AminoPVA USPIO NPs invaded three-dimensional spheroids of both HT-29 and Caco-2 cells, whereas oleic acid-coated USPIO NPs could only invade Caco-2 spheroids. Neither cationic aminoPVA USPIO NPs nor anionic oleic acid-coated USPIO NPs were transported at detectable levels across the tight CacoReady™ intestinal barrier model or the more permeable mucus-secreting CacoGoblet™ model.

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