Penetration of CdSe/ZnS quantum dots into differentiated vs undifferentiated Caco-2 cells.
BACKGROUND
Quantum dots (QDs) have great potential as fluorescent labels but cytotoxicity relating to extra- and intracellular degradation in biological systems has to be addressed prior to biomedical applications. In this study, human intestinal cells (Caco-2) grown on transwell membranes were used to study penetration depth, intracellular localization, translocation and cytotoxicity of CdSe/ZnS QDs with amino and carboxyl surface modifications. The focus of this study was to compare the penetration depth of QDs in differentiated vs undifferentiated cells using confocal microscopy and image processing.


RESULTS
Caco-2 cells were exposed to QDs with amino (NH2) and carboxyl (COOH) surface groups for 3 days using a concentration of 45 µg cadmium ml-1. Image analysis of confocal/multiphoton microscopy z-stacks revealed no penetration of QDs into the cell lumen of differentiated Caco-2 cells. Interestingly, translocation of cadmium ions onto the basolateral side of differentiated monolayers was observed using high resolution inductively coupled plasma mass spectrometry (ICP-MS). Membrane damage was neither detected after short nor long term incubation in Caco-2 cells. On the other hand, intracellular localization of QDs after exposure to undifferentiated cells was observed and QDs were partially located within lysosomes.


CONCLUSIONS
In differentiated Caco-2 monolayers, representing a model for small intestinal enterocytes, no penetration of amino and carboxyl functionalized CdSe/ZnS QDs into the cell lumen was detected using microscopy analysis and image processing. In contrast, translocation of cadmium ions onto the basolateral side could be detected using ICP-MS. However, even after long term incubation, the integrity of the cell monolayer was not impaired and no cytotoxic effects could be detected. In undifferentiated Caco-2 cells, both QD modifications could be found in the cell lumen. Only to some extend, QDs were localized in endosomes or lysosomes in these cells. The results indicate that the differentiation status of Caco-2 cells is an important factor in internalization and localization studies using Caco-2 cells. Furthermore, a combination of microscopy analysis and sensitive detection techniques like ICP-MS are necessary for studying the interaction of cadmium containing QDs with cells.
INM - Leibniz Institute for New Materials, Campus D2 2, 66123, Saarbrücken, Germany.
Henrike
H
Peuschel
INM - Leibniz Institute for New Materials, Campus D2 2, 66123, Saarbrücken, Germany.
Thomas
T
Ruckelshausen
INM - Leibniz Institute for New Materials, Campus D2 2, 66123, Saarbrücken, Germany.
Silke
S
Kiefer
INM - Leibniz Institute for New Materials, Campus D2 2, 66123, Saarbrücken, Germany.
Yuliya
Y
Silina
INM - Leibniz Institute for New Materials, Campus D2 2, 66123, Saarbrücken, Germany. annette.kraegeloh@leibniz-inm.de.
Annette
A
Kraegeloh
Differentiation; Human intestinal cells (Caco-2); Penetration depth; Quantum dots
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