Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods

Friedrich RP, Janko C, Poettler M, Tripal P, Zaloga J, Cicha I, Dürr S, Nowak J, Odenbach S, Slabu I, Liebl M, Trahms L, Stapf M, Hilger I, Lyer S, Alexiou C (2015)


Publication Type: Journal article

Publication year: 2015

Journal

Publisher: Dove Medical Press

Book Volume: 10

Pages Range: 4185-201

DOI: 10.2147/IJN.S82714

Abstract

Due to their special physicochemical properties, iron nanoparticles offer new promising possibilities for biomedical applications. For bench to bedside translation of super-paramagnetic iron oxide nanoparticles (SPIONs), safety issues have to be comprehensively clarified. To understand concentration-dependent nanoparticle-mediated toxicity, the exact quantification of intracellular SPIONs by reliable methods is of great importance. In the present study, we compared three different SPION quantification methods (ultraviolet spectrophotometry, magnetic particle spectroscopy, atomic adsorption spectroscopy) and discussed the shortcomings and advantages of each method. Moreover, we used those results to evaluate the possibility to use flow cytometric technique to determine the cellular SPION content. For this purpose, we correlated the side scatter data received from flow cytometry with the actual cellular SPION amount. We showed that flow cytometry provides a rapid and reliable method to assess the cellular SPION content. Our data also demonstrate that internalization of iron oxide nanoparticles in human umbilical vein endothelial cells is strongly dependent to the SPION type and results in a dose-dependent increase of toxicity. Thus, treatment with lauric acid-coated SPIONs (SEON(LA)) resulted in a significant increase in the intensity of side scatter and toxicity, whereas SEON(LA) with an additional protein corona formed by bovine serum albumin (SEON(LA-BSA)) and commercially available Rienso(®) particles showed only a minimal increase in both side scatter intensity and cellular toxicity. The increase in side scatter was in accordance with the measurements for SPION content by the atomic adsorption spectroscopy reference method. In summary, our data show that flow cytometry analysis can be used for estimation of uptake of SPIONs by mammalian cells and provides a fast tool for scientists to evaluate the safety of nanoparticle products.

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How to cite

APA:

Friedrich, R.P., Janko, C., Poettler, M., Tripal, P., Zaloga, J., Cicha, I.,... Alexiou, C. (2015). Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods. International Journal of Nanomedicine, 10, 4185-201. https://dx.doi.org/10.2147/IJN.S82714

MLA:

Friedrich, Ralf P., et al. "Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods." International Journal of Nanomedicine 10 (2015): 4185-201.

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