Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance

Cristina Riggio,1,* Maria Pilar Calatayud,2,* Clare Hoskins,3 Josephine Pinkernelle,4 Beatriz Sanz,2 Teobaldo Enrique Torres,2,5 Manuel Ricardo Ibarra,2,5 Lijun Wang,3 Gerburg Keilhoff,4 Gerardo Fabian Goya,2,5 Vittoria Raffa,1,6 Alfred Cuschieri1,3 1Institute of Life Science, Scuola Superiore Sant&...

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Autores principales:	Riggio C, Calatayud MP, Hoskins C, Pinkernelle J, Sanz B, Torres TE, Ibarra MR, Wang L, Keilhoff G, Goya GF, Raffa V, Cuschieri A
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2012
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/0ba8344a9bd64f98bbbf4d6b6e27c7cf
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id	oai:doaj.org-article:0ba8344a9bd64f98bbbf4d6b6e27c7cf
record_format	dspace
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Riggio C Calatayud MP Hoskins C Pinkernelle J Sanz B Torres TE Ibarra MR Wang L Keilhoff G Goya GF Raffa V Cuschieri A Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
description	Cristina Riggio,1,* Maria Pilar Calatayud,2,* Clare Hoskins,3 Josephine Pinkernelle,4 Beatriz Sanz,2 Teobaldo Enrique Torres,2,5 Manuel Ricardo Ibarra,2,5 Lijun Wang,3 Gerburg Keilhoff,4 Gerardo Fabian Goya,2,5 Vittoria Raffa,1,6 Alfred Cuschieri1,3 1Institute of Life Science, Scuola Superiore Sant&rsquo;Anna, Piazza Martiri della Libert&agrave;, Pisa, Italy; 2Instituto de Nanociencia de Arag&oacute;n, Universidad de Zaragoza. Mariano Esquillor, Zaragoza, Spain; 3IMSaT, Institute for Medical Science and Technology, University of Dundee, Dundee, Scotland; 4Otto-von-Guericke University, Institute of Biochemistry and Cell Biology, Magdeburg, Germany; 5Departamento de F&iacute;sica de la Materia Condensada, Facultad de Ciencias, Universidad de Zaragoza. Cerbuna 12, Zaragoza, Spain; 6Department of Biology, Universit&agrave; di Pisa, Pisa, Italy*These authors contributed equally to this workPurpose: It has been proposed in the literature that Fe3O4 magnetic nanoparticles (MNPs) could be exploited to enhance or accelerate nerve regeneration and to provide guidance for regenerating axons. MNPs could create mechanical tension that stimulates the growth and elongation of axons. Particles suitable for this purpose should possess (1) high saturation magnetization, (2) a negligible cytotoxic profile, and (3) a high capacity to magnetize mammalian cells. Unfortunately, the materials currently available on the market do not satisfy these criteria; therefore, this work attempts to overcome these deficiencies.Methods: Magnetite particles were synthesized by an oxidative hydrolysis method and characterized based on their external morphology and size distribution (high-resolution transmission electron microscopy [HR-TEM]) as well as their colloidal (Z potential) and magnetic properties (Superconducting QUantum Interference Devices [SQUID]). Cell viability was assessed via Trypan blue dye exclusion assay, cell doubling time, and MTT cell proliferation assay and reactive oxygen species production. Particle uptake was monitored via Prussian blue staining, intracellular iron content quantification via a ferrozine-based assay, and direct visualization by dual-beam (focused ion beam/scanning electron microscopy [FIB/SEM]) analysis. Experiments were performed on human neuroblastoma SH-SY5Y cell line and primary Schwann cell cultures of the peripheral nervous system.Results: This paper reports on the synthesis and characterization of polymer-coated magnetic Fe3O4 nanoparticles with an average diameter of 73 &plusmn; 6 nm that are designed as magnetic actuators for neural guidance. The cells were able to incorporate quantities of iron up to 2 pg/cell. The intracellular distribution of MNPs obtained by optical and electronic microscopy showed large structures of MNPs crossing the cell membrane into the cytoplasm, thus rendering them suitable for magnetic manipulation by external magnetic fields. Specifically, migration experiments under external magnetic fields confirmed that these MNPs can effectively actuate the cells, thus inducing measurable migration towards predefined directions more effectively than commercial nanoparticles (fluidMAG-ARA supplied by Chemicell). There were no observable toxic effects from MNPs on cell viability for working concentrations of 10 &micro;g/mL (EC25 of 20.8 &micro;g/mL, compared to 12 &micro;g/mL in fluidMAG-ARA). Cell proliferation assays performed with primary cell cultures of the peripheral nervous system confirmed moderate cytotoxicity (EC25 of 10.35 &micro;g/mL).Conclusion: These results indicate that loading neural cells with the proposed MNPs is likely to be an effective strategy for promoting non-invasive neural regeneration through cell magnetic actuation.Keywords: magnetic nanoparticle, actuator, migration, neural regeneration
format	article
author	Riggio C Calatayud MP Hoskins C Pinkernelle J Sanz B Torres TE Ibarra MR Wang L Keilhoff G Goya GF Raffa V Cuschieri A
author_facet	Riggio C Calatayud MP Hoskins C Pinkernelle J Sanz B Torres TE Ibarra MR Wang L Keilhoff G Goya GF Raffa V Cuschieri A
author_sort	Riggio C
title	Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
title_short	Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
title_full	Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
title_fullStr	Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
title_full_unstemmed	Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
title_sort	poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance
publisher	Dove Medical Press
publishDate	2012
url	https://doaj.org/article/0ba8344a9bd64f98bbbf4d6b6e27c7cf
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spelling	oai:doaj.org-article:0ba8344a9bd64f98bbbf4d6b6e27c7cf2021-12-02T07:28:37ZPoly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance1176-91141178-2013https://doaj.org/article/0ba8344a9bd64f98bbbf4d6b6e27c7cf2012-06-01T00:00:00Zhttp://www.dovepress.com/poly-l-lysine-coated-magnetic-nanoparticles-as-intracellular-actuators-a10216https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Cristina Riggio,1,* Maria Pilar Calatayud,2,* Clare Hoskins,3 Josephine Pinkernelle,4 Beatriz Sanz,2 Teobaldo Enrique Torres,2,5 Manuel Ricardo Ibarra,2,5 Lijun Wang,3 Gerburg Keilhoff,4 Gerardo Fabian Goya,2,5 Vittoria Raffa,1,6 Alfred Cuschieri1,3 1Institute of Life Science, Scuola Superiore Sant&rsquo;Anna, Piazza Martiri della Libert&agrave;, Pisa, Italy; 2Instituto de Nanociencia de Arag&oacute;n, Universidad de Zaragoza. Mariano Esquillor, Zaragoza, Spain; 3IMSaT, Institute for Medical Science and Technology, University of Dundee, Dundee, Scotland; 4Otto-von-Guericke University, Institute of Biochemistry and Cell Biology, Magdeburg, Germany; 5Departamento de F&iacute;sica de la Materia Condensada, Facultad de Ciencias, Universidad de Zaragoza. Cerbuna 12, Zaragoza, Spain; 6Department of Biology, Universit&agrave; di Pisa, Pisa, Italy*These authors contributed equally to this workPurpose: It has been proposed in the literature that Fe3O4 magnetic nanoparticles (MNPs) could be exploited to enhance or accelerate nerve regeneration and to provide guidance for regenerating axons. MNPs could create mechanical tension that stimulates the growth and elongation of axons. Particles suitable for this purpose should possess (1) high saturation magnetization, (2) a negligible cytotoxic profile, and (3) a high capacity to magnetize mammalian cells. Unfortunately, the materials currently available on the market do not satisfy these criteria; therefore, this work attempts to overcome these deficiencies.Methods: Magnetite particles were synthesized by an oxidative hydrolysis method and characterized based on their external morphology and size distribution (high-resolution transmission electron microscopy [HR-TEM]) as well as their colloidal (Z potential) and magnetic properties (Superconducting QUantum Interference Devices [SQUID]). Cell viability was assessed via Trypan blue dye exclusion assay, cell doubling time, and MTT cell proliferation assay and reactive oxygen species production. Particle uptake was monitored via Prussian blue staining, intracellular iron content quantification via a ferrozine-based assay, and direct visualization by dual-beam (focused ion beam/scanning electron microscopy [FIB/SEM]) analysis. Experiments were performed on human neuroblastoma SH-SY5Y cell line and primary Schwann cell cultures of the peripheral nervous system.Results: This paper reports on the synthesis and characterization of polymer-coated magnetic Fe3O4 nanoparticles with an average diameter of 73 &plusmn; 6 nm that are designed as magnetic actuators for neural guidance. The cells were able to incorporate quantities of iron up to 2 pg/cell. The intracellular distribution of MNPs obtained by optical and electronic microscopy showed large structures of MNPs crossing the cell membrane into the cytoplasm, thus rendering them suitable for magnetic manipulation by external magnetic fields. Specifically, migration experiments under external magnetic fields confirmed that these MNPs can effectively actuate the cells, thus inducing measurable migration towards predefined directions more effectively than commercial nanoparticles (fluidMAG-ARA supplied by Chemicell). There were no observable toxic effects from MNPs on cell viability for working concentrations of 10 &micro;g/mL (EC25 of 20.8 &micro;g/mL, compared to 12 &micro;g/mL in fluidMAG-ARA). Cell proliferation assays performed with primary cell cultures of the peripheral nervous system confirmed moderate cytotoxicity (EC25 of 10.35 &micro;g/mL).Conclusion: These results indicate that loading neural cells with the proposed MNPs is likely to be an effective strategy for promoting non-invasive neural regeneration through cell magnetic actuation.Keywords: magnetic nanoparticle, actuator, migration, neural regenerationRiggio CCalatayud MPHoskins CPinkernelle JSanz BTorres TEIbarra MRWang LKeilhoff GGoya GFRaffa VCuschieri ADove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2012, Iss default, Pp 3155-3166 (2012)

Poly-l-lysine-coated magnetic nanoparticles as intracellular actuators for neural guidance

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