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

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

Ralf P Friedrich,1 Christina Janko,1 Marina Poettler,1 Philipp Tripal,1 Jan Zaloga,1 Iwona Cicha,1 Stephan Dürr,1,2 Johannes Nowak,3 Stefan Odenbach,3 Ioana Slabu,4 Maik Liebl,4 Lutz Trahms,4 Marcus Stapf,5 Ingrid Hilger,5 Stefan Lyer,1 Christoph Alexiou1 1Department of Otorhinolaryngology...

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Autores principales: 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
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2015
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Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/328fb6642ce34b3484fe818bd3aba31f
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spelling oai:doaj.org-article:328fb6642ce34b3484fe818bd3aba31f2021-12-02T05:52:54ZFlow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods1178-2013https://doaj.org/article/328fb6642ce34b3484fe818bd3aba31f2015-06-01T00:00:00Zhttp://www.dovepress.com/flow-cytometry-for-intracellular-spion-quantification-specificity-and--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Ralf P Friedrich,1 Christina Janko,1 Marina Poettler,1 Philipp Tripal,1 Jan Zaloga,1 Iwona Cicha,1 Stephan Dürr,1,2 Johannes Nowak,3 Stefan Odenbach,3 Ioana Slabu,4 Maik Liebl,4 Lutz Trahms,4 Marcus Stapf,5 Ingrid Hilger,5 Stefan Lyer,1 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine, University hospital Erlangen, 2Department of Otorhinolaryngology, Head and Neck Surgery, Section of Phoniatrics and Pediatric Audiology, University hospital Erlangen, Erlangen, 3Technische Universität Dresden, Chair of Magnetofluiddynamics, Measuring and Automation Technology, Dresden, 4Physikalisch-Technische Bundesanstalt Berlin, Berlin, 5Department of Radiology, Division of Diagnostic and Interventional Radiology, Experimental Radiology, University hospital Jena, Jena, Germany Abstract: Due to their special physicochemical properties, iron nanoparticles offer new promising possibilities for biomedical applications. For bench to bedside translation of superparamagnetic 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 (SEONLA) resulted in a significant increase in the intensity of side scatter and toxicity, whereas SEONLA with an additional protein corona formed by bovine serum albumin (SEONLA-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. Keywords: low cytometry, side scatter, intracellular superparamagnetic iron oxide nanoparticles, quantification, spectroscopyFriedrich RPJanko CPoettler MTripal PZaloga JCicha IDürr SNowak JOdenbach SSlabu ILiebl MTrahms LStapf MHilger ILyer SAlexiou CDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2015, Iss default, Pp 4185-4201 (2015)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
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
Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
description Ralf P Friedrich,1 Christina Janko,1 Marina Poettler,1 Philipp Tripal,1 Jan Zaloga,1 Iwona Cicha,1 Stephan Dürr,1,2 Johannes Nowak,3 Stefan Odenbach,3 Ioana Slabu,4 Maik Liebl,4 Lutz Trahms,4 Marcus Stapf,5 Ingrid Hilger,5 Stefan Lyer,1 Christoph Alexiou1 1Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine, University hospital Erlangen, 2Department of Otorhinolaryngology, Head and Neck Surgery, Section of Phoniatrics and Pediatric Audiology, University hospital Erlangen, Erlangen, 3Technische Universität Dresden, Chair of Magnetofluiddynamics, Measuring and Automation Technology, Dresden, 4Physikalisch-Technische Bundesanstalt Berlin, Berlin, 5Department of Radiology, Division of Diagnostic and Interventional Radiology, Experimental Radiology, University hospital Jena, Jena, Germany Abstract: Due to their special physicochemical properties, iron nanoparticles offer new promising possibilities for biomedical applications. For bench to bedside translation of superparamagnetic 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 (SEONLA) resulted in a significant increase in the intensity of side scatter and toxicity, whereas SEONLA with an additional protein corona formed by bovine serum albumin (SEONLA-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. Keywords: low cytometry, side scatter, intracellular superparamagnetic iron oxide nanoparticles, quantification, spectroscopy
format article
author 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
author_facet 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
author_sort Friedrich RP
title Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
title_short Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
title_full Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
title_fullStr Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
title_full_unstemmed Flow cytometry for intracellular SPION quantification: specificity and sensitivity in comparison with spectroscopic methods
title_sort flow cytometry for intracellular spion quantification: specificity and sensitivity in comparison with spectroscopic methods
publisher Dove Medical Press
publishDate 2015
url https://doaj.org/article/328fb6642ce34b3484fe818bd3aba31f
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