XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
248
T2: P–33
Even low doses of iron oxides nanoparticles may disturb the elemental
composition of selected body organs – preliminary results obtained
for rats using TXRF spectroscopy
Agnieszka Skoczeń
1
, Katarzyna Matusiak
1
, Zuzanna Setkowicz
2
,
Aldona Kubala-Kukus
3
, Ilona Stabrawa
3
, Małgorzata Ciarach
2
,
Krzysztof Janeczko
2
, and Joanna Chwiej
1
1
Faculty of Physics and Applied Computer Science, AGH University of Science and Technology,
al. Mickiewicza 30, PL-30059 Krakow, Poland, e-mail: Agnieszka.Skoczen@fis.agh.edu.pl
2
Institute of Zoology, Jagiellonian University, Gronostajowa 9, PL-30387 Krakow, Poland
3
Laboratory of X-ray Methods, The Jan Kochanowski University, Świętokrzyska 15, PL-25406
Kielce, Poland
Despite the possibilities offered by advances in the field of nanotechnology, the use of nano-
objects in medicine is not possible without prior examination of their impact on the living
organisms [1–3]. In vivo studies carried out on animals are one of the first steps towards it. They
allow to determine organs of nanoparticles (NPs) accumulation as well as evaluate structural and
functional changes caused by these nano-objects.
This investigation aimed at evaluation of biodistribution and potential toxic effects of NPs
on rats. PEG-coated magnetic iron oxides nanoparticles (IONPs) with the diameter of 30 nm
were suspended in 15% mannitol solution and intravenously injected to animals. The applied
dose of nanoparticles was approximately two orders of magnitude lower than doses used in
medical applications [4, 5]. The dynamics of IONPs induced elemental changes was studied for
the following body organs: kidneys, heart, spleen, brain and muscles. The elemental anomalies
of the liver were the subject of the earlier investigation. To achieve the purpose of the study, the
organs taken from rats 2 hours, 24 hours and 7 days from nanoparticles injection were
examined.
The investigation required the use of higly-sensitive analytical technique. Therefore, total
reflection X-ray fluorescence (TXRF) spectroscopy was applied in the study. With low detection
limits (ppm-ppb) TXRF method enabled us to examine the changes in Fe, Cu, Zn and Ca
concentration occurring in the mentioned rat organs after administration of relatively low doses
of IONPs.
Obtained results revealed short-term accumulation of IONPs in liver (the result from the
previous study) and kidneys of animals treated with NPs. Furthermore, the analysis of
experimental data showed that even such low dose of IONPs can cause toxic effects manifesting
mainly by abnormalities in Ca, Cu and Zn content in the kidney, heart and muscles. Such
changes in elemental concentration may result from the oxidative stress caused by the IONPs-
induced injury as well as triggering of mechanisms responsible for Fe homeostasis in organism.
Keywords: nanoparticles toxicity; TXRF; elemental analysis; rat organs
References
[1] B. Fadeel, A.E. Garcia-Bennett, Adv. Drug Deliv. Rev. 62 (2010) 362.
[2] Q. a Pankhurst, J. Connolly, S.K. Jones, J. Dobson, J. Phys. D. Appl. Phys., 36 (2003) R167.
[3] A. Dhawan and V. Sharma, Anal. Bioanal. Chem. 398 (2010) 589.
[4] B. Chertok, B.A. Moffat, A.E. David, F. Yu, C. Bergemann, B. D. Ross, V.C. Yang, Biomaterials 29
(2008) 487.
[5] T.K. Jain, M.K. Reddy, M.A. Morales, D.L. Leslie-Pelecky,. Labhasetwar, Mol. Pharm. 5 (2008) 316.
XIV
h
International Conference on Molecular Spectroscopy, Białka Tatrzańska 2017
249
T2: P–34
The use of multivariate methods for determination of the elemental
markers of exposure to iron oxides nanoparticles
Joanna Chwiej
1
, Agnieszka Skoczen
1
, Katarzyna Matusiak
1
, Zuzanna Setkowicz
2
,
Krzysztof Janeczko
2
, Aldona Kubala-Kukus
3,4
, and Ilona Stabrawa
3,4
1
AGH University of Science and Technology, Faculty of Physics and Applied Computer Science,
Mickiewicza 30, 30-059 Krakow, Poland, e-mail: Joanna.Chwiej@fis.agh.edu.pl
2
Jagiellonian University, Institute of Zoology, Gronostajowa 9, 30-387 Krakow, Poland
3
The Jan Kochanowski University, Institute of Physics, Swietokrzyska 15, 25-406 Kielce, Poland
4
The Holly Cross Cancer Center, Artwinskiego 3, 25-734 Kielce, 25-406 Kielce, Poland
Multivariate analysis consists of a collection of methods that can be used when several
measurements are made simultaneously on each individual or object in one or more samples [1].
Historically, many of applications of multivariate techniques have been performed in behavioral
and biological sciences. Nowadays, the interest in these methods has spread to numerous other
fields of investigation [1]. Cluster analysis, principal components analysis and discriminant
analysis are the three primary modern multivariate techniques [2]. The main goals of their use
are: reduction and simplification of the data structure, analysis of the relationships between
variables and grouping of the observations into groups defined a priori or separated during
analysis [1, 2].
NPs are comparable to the cellular components and biological molecules in size and
therefore they can easily cross the natural barriers protecting human body and induce adverse
health effects [3]. Although potential toxicities of NPs and properties driving such toxic effects
are still very badly understood, the development of these novel materials for biomedical
applications is undergoing a dramatic expansion [4].
In this work the usefulness of the two methods of multivariate analysis, namely cluster and
discriminant analysis, for identification of the elemental markers of exposure to iron oxides
nanoparticles (IONPs) action was examined. To achieve this purpose, the information about the
elemental composition of selected body organs taken from rats treated with IONPs and naive
controls was analyzed. The PEG-coated magnetic IONPs were intravenously injected to animals
and the highly sensitive method of total reflection X-ray fluorescence was used for the
multielemental analysis of liver, spleen, kidneys, brain, heart and muscles.
The cluster analysis was used for unsupervised classification of cases. In turn, the
discriminant analysis was applied in order to group observations into predefined groups, but
mainly for determination of elements (markers of exposure) playing the most important role in
the differentiation of these groups.
Keywords: iron oxides nanoparticles; TXRF; elemental analysis; multivariate methods;
cluster and discriminant analysis
References
[1] A.C. Rechner, [in:] Methods of multivariate analysis. Chichester, JohnWiley&Sons, 2002.
[2] J.R. Kettenring, J. Classif. 23 (2006) 3.
[3] A. Dhawan, V. Sharma, Anal. Bioanal. Chem. 398 (2010) 589.
[4] H. C. Fischer, W. C. Chan, Curr. Op. Biotech. 18 (2007) 565.
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