Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections

V Graz&uacute;,1 AM Silber,2 M Moros,1 L As&iacute;n,1 TE Torres,1,3,5 C Marquina,3,4 MR Ibarra,1,3 GF Goya1,31Instituto de Nanociencia de Arag&oacute;n (INA), Universidad de Zaragoza, Zaragoza, Spain; 2Departamento de Parasitologia, Instituto de Ci&ecirc;ncias Biom&a...

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Autores principales:	Graz&uacute; V, Silber AM, Moros M, As&iacute;n L, Torres TE, Marquina C, Ibarra MR, Goya GF
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Lenguaje:	EN
Publicado:	Dove Medical Press 2012
Materias:	Medicine (General) R5-920
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spelling	oai:doaj.org-article:0075e5b503294b8db1acc4f0fe8f21122021-12-02T01:16:05ZApplication of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections1176-91141178-2013https://doaj.org/article/0075e5b503294b8db1acc4f0fe8f21122012-10-01T00:00:00Zhttp://www.dovepress.com/application-of-magnetically-induced-hyperthermia-in-the-model-protozoa-a11202https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013V Graz&uacute;,1 AM Silber,2 M Moros,1 L As&iacute;n,1 TE Torres,1,3,5 C Marquina,3,4 MR Ibarra,1,3 GF Goya1,31Instituto de Nanociencia de Arag&oacute;n (INA), Universidad de Zaragoza, Zaragoza, Spain; 2Departamento de Parasitologia, Instituto de Ci&ecirc;ncias Biom&eacute;dicas, Universidade de S&atilde;o Paulo, S&atilde;o Paulo, Brazil; 3Departamento de F&iacute;sica de la Materia Condensada, Facultad de Ciencias, Universidad de Zaragoza, Zaragoza, Spain; 4Instituto de Ciencia de Materiales de Arag&oacute;n (ICMA), CSIC, Universidad de Zaragoza, Zaragoza, Spain; 5Laboratorio de Microscop&iacute;as Avanzadas (LMA), Universidad de Zaragoza, Zaragoza, SpainBackground: Magnetic hyperthermia is currently a clinical therapy approved in the European Union for treatment of tumor cells, and uses magnetic nanoparticles (MNPs) under time-varying magnetic fields (TVMFs). The same basic principle seems promising against trypanosomatids causing Chagas disease and sleeping sickness, given that the therapeutic drugs available have severe side effects and that there are drug-resistant strains. However, no applications of this strategy against protozoan-induced diseases have been reported so far. In the present study, Crithidia fasciculata, a widely used model for therapeutic strategies against pathogenic trypanosomatids, was targeted with Fe3O4 MNPs in order to provoke cell death remotely using TVMFs.Methods: Iron oxide MNPs with average diameters of approximately 30 nm were synthesized by precipitation of FeSO4 in basic medium. The MNPs were added to C. fasciculata choanomastigotes in the exponential phase and incubated overnight, removing excess MNPs using a DEAE-cellulose resin column. The amount of MNPs uploaded per cell was determined by magnetic measurement. The cells bearing MNPs were submitted to TVMFs using a homemade AC field applicator (f = 249 kHz, H = 13 kA/m), and the temperature variation during the experiments was measured. Scanning electron microscopy was used to assess morphological changes after the TVMF experiments. Cell viability was analyzed using an MTT colorimetric assay and flow cytometry.Results: MNPs were incorporated into the cells, with no noticeable cytotoxicity. When a TVMF was applied to cells bearing MNPs, massive cell death was induced via a nonapoptotic mechanism. No effects were observed by applying TVMF to control cells not loaded with MNPs. No macroscopic rise in temperature was observed in the extracellular medium during the experiments.Conclusion: As a proof of principle, these data indicate that intracellular hyperthermia is a suitable technology to induce death of protozoan parasites bearing MNPs. These findings expand the possibilities for new therapeutic strategies combating parasitic infection.Keywords: magnetic hyperthermia, magnetic nanoparticles, trypanosomatids, Crithidia fasciculataGraz&uacute; VSilber AMMoros MAs&iacute;n LTorres TEMarquina CIbarra MRGoya GFDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2012, Iss default, Pp 5351-5360 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Graz&uacute; V Silber AM Moros M As&iacute;n L Torres TE Marquina C Ibarra MR Goya GF Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
description	V Graz&uacute;,1 AM Silber,2 M Moros,1 L As&iacute;n,1 TE Torres,1,3,5 C Marquina,3,4 MR Ibarra,1,3 GF Goya1,31Instituto de Nanociencia de Arag&oacute;n (INA), Universidad de Zaragoza, Zaragoza, Spain; 2Departamento de Parasitologia, Instituto de Ci&ecirc;ncias Biom&eacute;dicas, Universidade de S&atilde;o Paulo, S&atilde;o Paulo, Brazil; 3Departamento de F&iacute;sica de la Materia Condensada, Facultad de Ciencias, Universidad de Zaragoza, Zaragoza, Spain; 4Instituto de Ciencia de Materiales de Arag&oacute;n (ICMA), CSIC, Universidad de Zaragoza, Zaragoza, Spain; 5Laboratorio de Microscop&iacute;as Avanzadas (LMA), Universidad de Zaragoza, Zaragoza, SpainBackground: Magnetic hyperthermia is currently a clinical therapy approved in the European Union for treatment of tumor cells, and uses magnetic nanoparticles (MNPs) under time-varying magnetic fields (TVMFs). The same basic principle seems promising against trypanosomatids causing Chagas disease and sleeping sickness, given that the therapeutic drugs available have severe side effects and that there are drug-resistant strains. However, no applications of this strategy against protozoan-induced diseases have been reported so far. In the present study, Crithidia fasciculata, a widely used model for therapeutic strategies against pathogenic trypanosomatids, was targeted with Fe3O4 MNPs in order to provoke cell death remotely using TVMFs.Methods: Iron oxide MNPs with average diameters of approximately 30 nm were synthesized by precipitation of FeSO4 in basic medium. The MNPs were added to C. fasciculata choanomastigotes in the exponential phase and incubated overnight, removing excess MNPs using a DEAE-cellulose resin column. The amount of MNPs uploaded per cell was determined by magnetic measurement. The cells bearing MNPs were submitted to TVMFs using a homemade AC field applicator (f = 249 kHz, H = 13 kA/m), and the temperature variation during the experiments was measured. Scanning electron microscopy was used to assess morphological changes after the TVMF experiments. Cell viability was analyzed using an MTT colorimetric assay and flow cytometry.Results: MNPs were incorporated into the cells, with no noticeable cytotoxicity. When a TVMF was applied to cells bearing MNPs, massive cell death was induced via a nonapoptotic mechanism. No effects were observed by applying TVMF to control cells not loaded with MNPs. No macroscopic rise in temperature was observed in the extracellular medium during the experiments.Conclusion: As a proof of principle, these data indicate that intracellular hyperthermia is a suitable technology to induce death of protozoan parasites bearing MNPs. These findings expand the possibilities for new therapeutic strategies combating parasitic infection.Keywords: magnetic hyperthermia, magnetic nanoparticles, trypanosomatids, Crithidia fasciculata
format	article
author	Graz&uacute; V Silber AM Moros M As&iacute;n L Torres TE Marquina C Ibarra MR Goya GF
author_facet	Graz&uacute; V Silber AM Moros M As&iacute;n L Torres TE Marquina C Ibarra MR Goya GF
author_sort	Graz&uacute; V
title	Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
title_short	Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
title_full	Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
title_fullStr	Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
title_full_unstemmed	Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections
title_sort	application of magnetically induced hyperthermia in the model protozoan crithidia fasciculata as a potential therapy against parasitic infections
publisher	Dove Medical Press
publishDate	2012
url	https://doaj.org/article/0075e5b503294b8db1acc4f0fe8f2112
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Application of magnetically induced hyperthermia in the model protozoan Crithidia fasciculata as a potential therapy against parasitic infections

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