Trinity College Dublin, the University of Dublin, Ireland · Trinity College Dublin, the University...
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Transcript of Trinity College Dublin, the University of Dublin, Ireland · Trinity College Dublin, the University...
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Table of Contents
MULTIFUN Consortium ............................................................................................. i
Dissemination preface ............................................................................................. 1
WMIC 2012 Workshop ............................................................................................ 5
Workshop Program ....................................................................................................................... 8
(I) Advances in imaging and diagnostics using nanotechnological tools ....................................................................... 9
(II) Advances in imaging and theranostics using nanotechnological tools .................................................................. 17
(III) Cluster: Targeted nanopharmaceuticals and diagnostics ..................................................................................... 25
(IV) Imaging cell and tissue interaction with nanomaterials ...................................................................................... 30
EuroNanoForum 2013 ........................................................................................... 37
Nanomedicine at EuroNanoForum 2013 ...................................................................................... 38
Highlights I: Nanomedicine: Technology Platforms and Breakthrough Projects (ENF Workshop 7) ............................ 39
Highlights II: Market Strategies in the Medical Device Industry (ENF Workshop 8) .................................................... 42
Final Multifun Workshop 2015 .............................................................................. 45
Workshop program ..................................................................................................................... 46
Workshop highlights ................................................................................................................................................. 51
Workshop organising committee .............................................................................................................................. 52
Acknowledgements ............................................................................................... 54
Funding ................................................................................................................. 54
Appendix 1 – Final Multifun workshop 2015, complete booklet ............................ 55
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Stephanie!#,/-8,+9 is the )=\!>&;!U*/&;,$!*C!<,N$%BG!98, took the initiative to set-up the Pepric and now has 5 years’
experience in the business development and financial aspects of the company. Previous to Pepric, she prepared and
assisted spin-off projects at Imec as Venture Development Manager. During her PhD she worked at several international
particle accelerator facilities (France, US, Japan) on nuclear magnetic resonance techniques. This offered her the nec-
essary background for a clear understanding of the technological principles of MRI, PET and SPECT. Dr. Teughels has
obtained postgraduate degrees in Corporate Finance (2006), Business Administration (2005), and nuclear physics (PhD,
2001).
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rect and selective detection method pEPR (particle Elec-
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37
EuroNanoForum 2013 At the 2013 EuroNanoForum (ENF) conference, the MULTIFUN project made a major contribution to the event, through the Irish project partner Trinity College Dublin, to bridging the gap between the world of medical research/innovation and that of the nanotechnology stakeholders, by collaborating to organize two work-‐shops entirely dedicated to the nanomedicine. The ENF was a great opportunity for the nanomedicine communi-‐ty to emphasize to a large panel of research, industry and public authority representatives, the huge potential of nanotechnology as an enabling technology for medical applications. With participation at ENF supported by the European Technology Platform on Nanomedicine (ETP Nanomedicine), the nanomedicine field was widely rep-‐resented and specific needs and requirements for the successful implementation of nanotechnologies in the health sector were highlighted.
The ENF 2013 conference, organized by Enterprise Ireland, was held in Dublin in June 2013 during the Irish presidency of the European Union under the motto “Nanotechnology Innovation: From research to commer-‐cialization, the bridge to Horizon 2020”. The event was a tremendous success, attracting nearly 1,500 active members of the nanotechnology community from 50 countries around the world. Over 140 high-‐level speakers presented at the plenaries, sessions and workshops. Nanotechnology developments and commercialization success stories were also visible at the coinciding Nano-‐tech Europe exhibition. The event was supported by the European Commission and its Industrial Technologies programme, and co-‐organised by Enterprise Ireland and Spinverse Ltd.
The focus of the conference was the commercializa-‐tion of nanotechnology, exploiting its potential for new applications, pushing it from an enabling technology through to use in end products. Industrial and research organizations such as BASF, Shell, Ottobock, Nanobiotix, Intel, Philips Healthcare, Airbus, VTT, Fraunhofer, Max-‐Planck Institute, CRANN, Tyndall, MSSI and NanoStart, initiated a thriving discussion about the future of nano-‐technology, its economic and technological impact on
European growth and the commercialization challenges of nanoproducts. Speakers agreed that understanding of industrial needs, focused R&D and suitable funding in-‐struments are required, as is the identification of areas where nanotechnology is most likely to have an impact.
The Nanotech Europe exhibition completed the con-‐ference. Here, 70 innovative organizations from 20 coun-‐tries like Austria, the UK, Germany, Denmark, Finland, France, the Czech Republic, Switzerland and the Nether-‐lands showcased their ground-‐breaking innovations. In the meantime, a brokerage day on June 20th saw 50 par-‐allel meetings take place, sparking new businesses, pro-‐ject consortia and cooperative activities for technology transfer.
With Horizon 2020 beginning in 2014, the confer-‐ence offered a unique chance to look at how nanotech-‐nology will fit into the new structure’s key priority areas of excellent science, industrial leadership and societal challenges. “Today, as demonstrated in ENF 2013, nano-‐technology applications can be found in all areas of in-‐dustrial technologies”, commented Dr. Herbert von Bose, Director of the Industrial Technologies Programme of DG Research and Innovation at the European Commission. “Still, a lot more needs to be done, especially in fastening the pace of industrial innovation. We are looking forward to continuing our work within the next seven-‐year pro-‐gramme of Horizon 2020, following the principles of safe, responsible and sustainable nanotechnology develop-‐ment and ensuring the competitiveness and growth of European industries”.
The central role of nanomedicine within the sphere of applied nanotechnology was one of the key topics at the conference; and this point was emphasized by the award for Best Research Project going to the EU nano-‐medicine project, SONODRUGS. The project aims to de-‐velop novel technologies for drug delivery to enable lo-‐calized treatment of cardiovascular disease and cancer. For patients, this could mean a treatment with fewer side effects and burdens, in addition to reduced post-‐intervention recovery times.
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2015 workshop
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february23-252015
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welcome
#multifunworkshop
During the last four years the MULTIFUN consortium has focused its activity on the development and validation of new systems based upon minimal invasive nanotechnology for the early and selective detection and elimination of breast and pancreatic cancer. The project has successfully produced multifuntionalised magnetic iron oxide nanoparticles (MNP) that combine diagnostic and therapeutic features against these two types of cancer.
The therapeutic approach developed within the project includes a synergistic effect between the therapeutic effect produced by magnetic hyperthermia and that due to the intracellular drug delivery selectively targeted to tumour cells.
Some of the designed formulations have proven their efficiency, safety and non-toxicicty in in vivo models, making them promising candidates to produce new nanomedicines against breast and pancreatic cancer.
The Madrid Institute for Advanced Studies in Nanoscience (IMDEA Nanociencia) is responsible of the scientific coordination of the project. Atos Spain S.A is the Administrative Coordinator and Contract Manager.
Prof. Rodolfo Miranda, MULTIFUN Scientific Coordinator &
IMDEA Nanociencia Director
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Monday 23 February 08.30 - 09.30 Registration
09.30 - 10:00 Opening session
Prof. Rodolfo Miranda, (MULTIFUN Scientific Coordina-tor & IMDEA Nanociencia Director)
Session 1: Introduction and Perspectives: Cancer Science
Chairperson: Prof. Rodolfo Miranda, IMDEA Nanociencia
10:00 - 10:45 Keynote speaker: Dr. Manuel Hidalgo, Centro Nacional de Investiga-ciones Oncológicas
“Nanodrugs: Development and Applications to Pan-creas Cancer Treatment”
10:45 - 11:30 Keynote speaker: Dr. José Antonio López, Instituto de Investigación Hospital 12 de Octubre
“An overview of cancer and (druggable-) immune response”
11:30 - 12:00 COFFEE BREAK
Session 2: Nanotoxicity, Biodistribution, Biodegradation
Chairperson: Prof. Yuri Volkov, Centre for Research on Adaptive Nanostructures, Trinity College Dublin
12:00 - 12:40 Keynote speaker: Prof. Florence Gazeau, Université Paris-Diderot-CNRS
“Ageing and biotransformation of nanomagnets in the body: How to conciliate therapeutic efficiency and safe life-cycle?”
12:40 - 13:30 Round table on regulation for manufacturing and biomedical applications of nanomaterials: safety on nanohandling.
Members: Prof. África González-Fernández Instituto de Investigaciones Biomédicas de Vigo - Universidad de Vigo, Prof. Florence Gazeau Université Paris-Diderot-CNRS, Prof. Yuri Volkov Trinity College Dublin
Moderator: Dr. Pilar Calvo Pharmamar
13:30 - 15:00 LUNCH BREAK
15:00 - 16:40 Poster session
Chairperson: Dr. Domingo F. Barber, Centro Nacional de Biotecnología-CSIC
16:40 - 17:10 Invited speaker: Prof. África González-Fernández, (Instituto de Investigaciones Biomédicas de Vigo - Uni-versidad de Vigo)
“Safety of nanomaterials: Immunotoxicology and inter-action between Nanomaterials and human proteins”
17:10 - 17:30 Mr. Vladimir Mulens, Centro Nacional de Biotecnología-CSIC
“Effect of Polyethylenimine-coated SPIONs on mac-rophage activation and podosome dynamics”
17:30 - 17:50 Dr. Lucía Gutiérrez, Instituto de Ciencia de Materiales de Madrid-CSIC
“Tracking magnetic nanoparticles in tissues by AC magnetic susceptibility: Recent achievements and future challenges”
17:50 - 18:10 Dr. Francisco Javier Chichón, Centro Nacional de Biotecnología-CSIC
“Soft X-ray tomography as quantitative tool to deci-pher the interaction between DMSA-SPION and MCF7 cancer cells”
18:10 - 18:30 Mr. K. Crosbie-Staunton, Trinity College Dublin “Multiparametric High Throughput Methods for Mag-
netic Iron Oxide Nanoparticle Lead Selection”
Tuesday 24 February Session 3: NanoDiagnostic
Chairperson: Prof. René Botnar, King’s College London
09:30 - 10:15 Keynote speaker: Prof. Kannan Krishnan, University of Washington
“Tracer development critical for translational medical applications of Magnetic Particle Imaging”
10:15 - 10:35 Prof. Yung-Ya Lin, University California, Los Angeles “Early Detection of Pancreatic Cancers & Brain
Tumors by MR Molecular Imaging”
10:35 - 10:55 Dr. F. Herranz, Centro Nacional de Investigaciones Car-diovasculares
“Parallel multifunctionalisation of Nanoparticles: A One-Step Modular Approach for in vivo Imaging”
10:55 - 11:15 Dr. Rocío Costo, Instituto de Ciencia de Materiales de Madrid-CSIC
“Improving magnetic properties of ultrasmall magnetic nanoparticles by biocompatible coatings”
10:15 - 11:30 Ms. Yurena Luengo, Instituto de Ciencia de Materiales de Madrid-CSIC
“Synthesis and characterization of bimetallic mag-netic oxides obtained by oxidative precipitation”
11:30 - 12:00 COFFEE BREAK
Chairperson: Dr. Jose Courty, CRRET-CNRS
12:00 - 12:20 Dr. Marzia Marciello, Instituto de Ciencia de Materiales de Madrid-CSIC
“Coating strategies for the application of magnetic nanoparticles in cancer diagnosis and therapy”
12:20 - 12:40 Dr. Wim Busing, FEI “Beyond the frontiers of the cell: unveiling the wonders
of nature at a molecular level”
12:40 - 13:30 Round table on entrepreneurship: from results to industry.
Members: Dr. Stephanie Teughels (CEO at Pepric), Dr. Rafael Ferritto (Co-founder and General Manager
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at NanoInnova Technologies), Ms. Cecilia Hernández (Head of Department Health, Bioeconomy, Climate and Natural Resources, Centre for Industrial Technological Development); Mr. Eduardo Díaz (Head of Unit New Technology Based Firms Unit, Fundación para el Cono-cimiento madri+d)
Moderator: Mr. Bonifacio Vega (Technology Transfer and Business Development Manager at IMDEA Nanociencia)
13:30 - 15:00 LUNCH BREAK
15:00 - 16:40 Poster session
Chairperson: Dr. Aitziber L. Cortajarena, IMDEA Nano-ciencia
16:40 - 17:10 Invited speaker: Dr Gray Kueberuwa, Institute of Cancer Sciences, University of Manchester
“Chimeric Antigen Receptor T-cells for Cancer Therapy”
17:10 - 17:30 Dr. Antonio Benayas, University of Quebec “At the Fluorescent Nanoprobes´ Frontiers: Multifunc-
tionality for Bio-Imaging and T-Sensing”
17:30 - 17:50 Dr. Ángel Millán, Instituto de Ciencias Materiales de Aragón
“Joining time-resolved thermometry and magnetic-induced heating in a single nanoparticle”
20:30 NETWORKING DINNER
Wednesday 25 February Session 3: NanoTherapy
Chairperson: Dr. Mª del Puerto Morales, Instituto de Ciencia de Materiales de Madrid-CSIC
09:30 - 10:15 Keynote speaker: Prof. Ingrid Hilger, University Hos-pital Jena
“Magnetic hyperthermia, a promising tool for the minimal-invasive treatment of tumors”
10:15 - 10:35 Dr. A. Roig, Instituto de Ciencia de Materiales de Bar-celona-CSIC
“Encapsulation of VEGF165 in magnetic PLGA nano-capsules for potential local delivery and bioactivity into human brain endothelial cells”
10:35 - 10:55 Mr. D. Cabrera, IMDEA Nanociencia “Influence of nanoparticle size and field frequency on
the concentration dependence of magnetic heating”
10:55 - 11:15 Ms. Blanca del Rosal, Universidad Autónoma de Madrid “Intratumoral thermal reading during photothermal
therapy by multifunctional fluorescent nanoparticles”
11:15 - 11:35 Prof. Nguyen TK Thanh, University College London “Magnetic nanoparticle-based therapeutic agents for
thermo-chemotherapy treatment of cancer”
11.35 -12:00 COFFEE BREAK
Chairperson: Dr. Sara Trabulo, Barts Cancer Institute
12:00 - 12:20 Dr. Cristina Fornaguera, Instituto de Química Avanzada de Cataluña-CSIC
“Nano-emulsion templating: a versatile technology to prepare multifunctional polymeric nanoparticles for advanced biomedical applications”
12:20 - 12.40 Dr. Anna Aviñó, Instituto de Química Avanzada de Cataluña-CSIC
“G-quadruplex aptamers with therapeutic applications”
12:40 - 13:30 Round table on nanotherapy: small treats different.
Members: Prof. Ingrid Hilger (University Hospital of Jena), Dr. Domingo F. Barber (Centro Nacional de Biotecnología-CSIC), Dr. Petra Gener (Hospital Vall d’Hebron)
Moderator: Dr. Aitziber L. Cortajarena, IMDEA Nano-ciencia
13:30-15:00 LUNCH BREAK
Chairperson: Dr. Pilar Calvo, Pharmamar
15:00 - 15:30 Invited Speaker: Dr. Petra Gener, Hospital Vall d’Hebron “Targeted Drug Delivery Systems against Cancer Stem
Cells”
15:30 - 15:50 Ms. Alicia Soler Cantón, Delft University of Technology “Development of gene-expressing liposomes as drug
delivery systems”
15:50 - 16:10 Dr. Laura Gallego-Yerga, Universidad de Sevilla “Self-assembling nanoparticles based on calixarene-
cyclodextrin heterodimers for targeted delivery and controlled release of docetaxel”
16:10 - 16:30 Mr. Gustavo da Silva, Instituto de Ciencia de Materiales de Madrid-CSIC
“Design of biocompatible magnetic nanoplatforms for potential platinum-based drug delivery applications”
16:30 - 17:00 CLOSING REMARKS
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ORAL contributions
Monday 23rd (morning session)
1. Nanodrugs Development and Applica-tions to Pancreas Cancer, Dr. Manuel Hidalgo, Centro Nacional de Investigaciones Oncológicas
2. An overview of cancer and (drugabble-) Immune response, Dr. José Antonio López, Instituto de Investigación Hospital 12 de Octubre
3. Ageing and biotransformation of nanomagnets in the body: How to con-ciliate therapeutic efficiency and safe life-cycle, Prof. Florence Gazeau, U. Paris Diderot-CNRS
Monday 23rd (afternoon session)
4. Safety of Nanomaterials: Immunotoxicol-ogy and interaction between Nanoma-terials and human proteins, Prof. Africa González Fernández, Instituto de Investiga-ciones Biomédicas de Vigo (UVIGO)
5. Effect of Polyethylenimine-coated SPIO-NS on macrophage activation and podo-some dynamics, Dr. Vladimir Mulens, Centro Nacional de Biotecnología-CSIC
6. Tracking magnetic nanoparticles in tissues by AC magnetic susceptibility: Recent achievements and future chal-lenges, Dr. Lucía Gutiérrez, Instituto de Ciencia de Materiales de Madrid-CSIC
7. Soft X-ray tomography as quantitative tool to decipher the interaction between DMSA-SPION and MCF7 cancer cells, Dr. Francisco Javier Chichón, Centro Nacional de Biotecnología-CSIC
8. Multiparamagnetic High Throughput Methods for Magnetic Iron Oxide Nano-particle Lead Selection, Mr. K. Crosbie-Staunton, Trinity College Dublin
Tuesday 24th (morning session)
9. Tracer development critical for transla-tional medical applications of Magnetic Particle Imaging, Prof. Kannan Krishnan, University of Washington
10. Early detection of Pancreatic Cancers & Brain Tumors by MR Molecular Imaging, Prof. Yung-Ya Lin, University California, Los Angeles
11. Parallel multifuncionalisation of nano-particles: A One-Step Modular Approach for in vivo Imaging, Dr. Fernando Herranz, Centro Nacional de Investigaciones Cardio-vasculares
12. Improving magnetic properties of ultras-mall magnetic nanoparticles by biocom-patible coatings, Dr. Rocío Costo, Instituto de Ciencia de Materiales de Madrid-CSIC
13. Synthesis and characterization of bimetal-lic magnetic oxides obtained by oxidative precipitation, Ms. Yurena Luengo, Instituto de Ciencia de Materiales de Madrid-CSIC
14. Coating strategies for the application of magnetic nanoparticles in cancer diagno-sis and therapy, Dr. Marzia Marciello, Insti-tuto de Ciencia de Materiales de Madrid-CSIC
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15. Beyond the frontiers of the cell: unveiling the wonders of nature at molecular level, Dr. Wim Busing, FEI
Tuesday 24th (afternoon session)
16. Chimeric Antigen Receptor T-cells for Cancer Therapy, Dr. Gray Kueberuwa, Institute of Cancer Sciences, University of Manchester
17. At the Fluorescent Nanoprobes’ Fron-tiers: Multifuncionality for Bio-Imaging and T-sensing, Dr. Antonio Benayas, University of Quebec
18. Joining time-resolved thermometry and magnetic-induced heating in a single nanoparticle, Dr. Ángel Millán, Instituto de Ciencias de Aragón
Wednesday 25th (morning session)
19. Magnetic hyperthermia, a promising tool for the minimal-invasive treatment of tumors, Prof. Ingrid Hilger, University Hospital Jena
20. Encapsulation of VEGF165 in magnetic PLGA nanocapsules for potential local delivery and bioactivity into human brain endothelial cells, Dr. A. Roig, Instituto de Ciencia de Materiales de Barcelona-CSIC
21. Influence of nanoparticle size and field frequency on the concentration depend-ence of magnetic heating, Mr. David Cabrera, Fundación IMDEA Nanociencia
22. Intratumoral thermal reading during photothermal therapy by multifunctional
fluorescent nanoparticles, Ms. Blanca del Rosal, Universidad Autónoma de Madrid
23. Magnetic nanoparticle-based therapeu-tic agents for thermo-chemotherapy treatment of cancer, Prof. Nguyen TK Thanh, University College London
24. Nano-emulsion templating: a versatile technology to prepare multifunctional polymeric nanoparticles for advanced biomedical applications, Dr. Cristina Fornaguera, Instituto de Química Avanzada de Cataluña-CSIC
25. G-quadruplex aptamers with therapeutic applications, Dr. Anna Aviñó, Instituto de Química Avanzada de Cataluña-CSIC
Wednesday 25th (afternoon session)
26. Targeted Drug Delivery Systems against Cancer Stem Cells, Dr. Petra Gener, Hos-pital Vall d’Hebron
27. Development of gene-expressing lipo-somes as drug delivery systems, Ms. Alicia Soler Cantón, Delft University of Technology
28. Self-assembling nanoparticles based on calixarene-cyclodextrin heterodimers for targeted delivery and controlled release of docetaxel, Dr. Laura Gallego Yerga, Universidad de Sevilla
29. Design of biocompatible magnetic nan-oplatforms for potential platinum-based drug delivery applications, Mr. Gustavo da Silva, Instituto de Ciencia de Materiales de Madrid-CSIC
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POSTER contributions
23rd and 24th afternoon session
P1. Interaction of multifunctional magnet-ic nanoparticles with biological mem-branes, Ms. María José Rodríguez, Centro Nacional de Biotecnología-CSIC
P2. Biodistribution of magnetic nano-particles evaluated by AC magnetic susceptibility in a murine model, Mr. Eduardo Lorente-Sorolla, Instituto de Ciencia de Materiales-CSIC
P3. C. elegans: an in vivo model to evaluate nanoparticles?, Dr. A. Roig, Instituto de Ciencia de Materiales de Barcelona-CSIC
P4. Iron quantification inside cells incubated with SPION by Soft-Xray Absorption Spectro-Tomography (SXAST), Mr. José Javier Conesa, Centro Nacional de Biotecnología-CSIC
P5. Following the transformation of mag-netic nanoparticles over time in mac-rophages by AC magnetic susceptibil-ity, Ms. Vanesa del Dedo, Instituto de Ciencia de Materiales de Madrid-CSIC
P6. pH-Dependent Switch for Anticancer Metallodrugs, Mr. Francisco Martínez, Fundación IMDEA Nanociencia
P7. Superparamagnetic properties of hydrothermally prepared CoFe2O4 as a function of size (6–10 nm) and coating (oleic/citric acid or TiO2), Dr. Daniel Nižnanský, Charles University in Prague
P8. Targeting cancer cells with photoac-tive silica nanoparticles, Ms. Wioleta Borzecka, University of Aveiro
P9. Combinational sensitization of tumor cells with two photosensitizers syn-ergistically enhances their photody-namic inactivation in vitro, Ms. And-rea Tabero, Universidad Autónoma de Madrid
P10. Synthesis Strategies of Single-Core Magnetic Nanoparticles, Ms. Helena Gavilán, Instituto de Ciencia de Materi-ales de Madrid-CSIC
P11. Silica encapsulation of magnetic nan-oparticles, Ms. Leonor de la Cueva, Fundación IMDEA Nanociencia
P12. Synthesis of hybrid magneto-plas-monic nanostructures based on Au nanorods and iron oxides nanoparti-cles, Mr. Jesús G. Ovejero, Universidad Complutense de Madrid
P13. Functionalized magnetic nanoparti-cles for cancer therapy, Dr. Antonio Aires, Fundación IMDEA Nanociencia
P14. Detection and Inhibition of Mutated GNAQ Gene using Spherical Nucleic Acids, Ms. Ana Latorre, Fundación IMDEA Nanociencia
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organizing committee
Chairperson:
Prof. Rodolfo Miranda (IMDEA Nanociencia)
Dr. Francisco J. Terán (IMDEA Nanociencia)
Dr. Adriele Prina Mello (TCD-CRANN)
Dr. César Mediavilla (ATOS)
advisory board committee Prof. Rene Botnar (KCL)
Dr. Pilar Calvo (Pharmamar)
Dr. Robert Clarke (UNIMAN)
Dr. Jose Courty (CRRET)
Dr. Domingo F. Barber (CSIC-CNB)
Prof. Christopher Heeschen (QMUL)
Dr. Aitziber L. Cortajarena (IMDEA Nanociencia)
Dr. Farouk Markos (UCC)
Dr. Mª del Puerto Morales (CSIC-ICMM)
Dr. Gorka Salas (IMDEA Nanociencia)
Prof. Yuri Volkov (TCD-CRANN)
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2015 workshop
final multifun
february23-252015
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2015 workshop
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welcome
#multifunworkshop
During the last four years the MULTIFUN
consortium has focused its activity on
the development and validation of new
systems based upon minimal invasive
nanotechnology for the early and selective
detection and elimination of breast and
pancreatic cancer. The project has
successfully produced multifuntionalised
magnetic iron oxide nanoparticles (MNP)
that combine diagnostic and therapeutic
features against these two types of cancer.
The therapeutic approach developed within
the project includes a synergistic effect
between the therapeutic effect produced
by magnetic hyperthermia and that due to
the intracellular drug delivery selectively
targeted to tumour cells.
Some of the designed formulations
have proven their efficiency, safety and
non-toxicicty in in vivo models, making
them promising candidates to produce
new nanomedicines against breast and
pancreatic cancer.
The Madrid Institute for Advanced Studies
in Nanoscience (IMDEA Nanociencia) is
responsible of the scientific coordination
of the project. Atos Spain S.A is the
Administrative Coordinator and Contract
Manager.
Prof. Rodolfo Miranda, MULTIFUN Scientific Coordinator &
IMDEA Nanociencia Director
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2015 workshop
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organizing committee
Chairperson:
Prof. Rodolfo Miranda (IMDEA Nanociencia)
Dr. Francisco J. Terán (IMDEA Nanociencia)
Dr. Adriele Prina Mello (TCD-CRANN)
Dr. César Mediavilla (ATOS)
advisory board committee Prof. Rene Botnar (KCL)
Dr. Pilar Calvo (Pharmamar)
Dr. Robert Clarke (UNIMAN)
Dr. Jose Courty (CRRET)
Dr. Domingo F. Barber (CSIC-CNB)
Prof. Christopher Heeschen (QMUL)
Dr. Aitziber L. Cortajarena (IMDEA Nanociencia)
Dr. Farouk Markos (UCC)
Dr. Mª del Puerto Morales (CSIC-ICMM)
Dr. Gorka Salas (IMDEA Nanociencia)
Prof. Yuri Volkov (TCD-CRANN)
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Monday 23 February 08.30 - 09.30 Registration
09.30 - 10:00 Opening session
Prof. Rodolfo Miranda, (MULTIFUN Scientific Coordina-tor & IMDEA Nanociencia Director)
Session 1: Introduction and Perspectives: Cancer Science
Chairperson: Prof. Rodolfo Miranda, IMDEA Nanociencia
10:00 - 10:45 Keynote speaker: Dr. Manuel Hidalgo, Centro Nacional de Investiga-ciones Oncológicas
“Nanodrugs: Development and Applications to Pan-creas Cancer Treatment”
10:45 - 11:30 Keynote speaker: Dr. José Antonio López, Instituto de Investigación Hospital 12 de Octubre
“An overview of cancer and (druggable-) immune response”
11:30 - 12:00 COFFEE BREAK
Session 2: Nanotoxicity, Biodistribution, Biodegradation
Chairperson: Prof. Yuri Volkov, Centre for Research on Adaptive Nanostructures, Trinity College Dublin
12:00 - 12:40 Keynote speaker: Prof. Florence Gazeau, Université Paris-Diderot-CNRS
“Ageing and biotransformation of nanomagnets in the body: How to conciliate therapeutic efficiency and safe life-cycle?”
12:40 - 13:30 Round table on regulation for manufacturing and biomedical applications of nanomaterials: safety on nanohandling.
Members: Prof. África González-Fernández Instituto de Investigaciones Biomédicas de Vigo - Universidad de Vigo, Prof. Florence Gazeau Université Paris-Diderot-CNRS, Prof. Yuri Volkov Trinity College Dublin
Moderator: Dr. Pilar Calvo Pharmamar
13:30 - 15:00 LUNCH BREAK
15:00 - 16:40 Poster session
Chairperson: Dr. Domingo F. Barber, Centro Nacional de Biotecnología-CSIC
16:40 - 17:10 Invited speaker: Prof. África González-Fernández, (Instituto de Investigaciones Biomédicas de Vigo - Uni-versidad de Vigo)
“Safety of nanomaterials: Immunotoxicology and inter-action between Nanomaterials and human proteins”
17:10 - 17:30 Mr. Vladimir Mulens, Centro Nacional de Biotecnología-CSIC
“Effect of Polyethylenimine-coated SPIONs on mac-rophage activation and podosome dynamics”
17:30 - 17:50 Dr. Lucía Gutiérrez, Instituto de Ciencia de Materiales de Madrid-CSIC
“Tracking magnetic nanoparticles in tissues by AC magnetic susceptibility: Recent achievements and future challenges”
17:50 - 18:10 Dr. Francisco Javier Chichón, Centro Nacional de Biotecnología-CSIC
“Soft X-ray tomography as quantitative tool to deci-pher the interaction between DMSA-SPION and MCF7 cancer cells”
18:10 - 18:30 Mr. K. Crosbie-Staunton, Trinity College Dublin “Multiparametric High Throughput Methods for Mag-
netic Iron Oxide Nanoparticle Lead Selection”
Tuesday 24 February Session 3: NanoDiagnostic
Chairperson: Prof. René Botnar, King’s College London
09:30 - 10:15 Keynote speaker: Prof. Kannan Krishnan, University of Washington
“Tracer development critical for translational medical applications of Magnetic Particle Imaging”
10:15 - 10:35 Prof. Yung-Ya Lin, University California, Los Angeles “Early Detection of Pancreatic Cancers & Brain
Tumors by MR Molecular Imaging”
10:35 - 10:55 Dr. F. Herranz, Centro Nacional de Investigaciones Car-diovasculares
“Parallel multifunctionalisation of Nanoparticles: A One-Step Modular Approach for in vivo Imaging”
10:55 - 11:15 Dr. Rocío Costo, Instituto de Ciencia de Materiales de Madrid-CSIC
“Improving magnetic properties of ultrasmall magnetic nanoparticles by biocompatible coatings”
10:15 - 11:30 Ms. Yurena Luengo, Instituto de Ciencia de Materiales de Madrid-CSIC
“Synthesis and characterization of bimetallic mag-netic oxides obtained by oxidative precipitation”
11:30 - 12:00 COFFEE BREAK
Chairperson: Dr. Jose Courty, CRRET-CNRS
12:00 - 12:20 Dr. Marzia Marciello, Instituto de Ciencia de Materiales de Madrid-CSIC
“Coating strategies for the application of magnetic nanoparticles in cancer diagnosis and therapy”
12:20 - 12:40 Dr. Wim Busing, FEI “Beyond the frontiers of the cell: unveiling the wonders
of nature at a molecular level”
12:40 - 13:30 Round table on entrepreneurship: from results to industry.
Members: Dr. Stephanie Teughels (CEO at Pepric), Dr. Rafael Ferritto (Co-founder and General Manager
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at NanoInnova Technologies), Ms. Cecilia Hernández (Head of Department Health, Bioeconomy, Climate and Natural Resources, Centre for Industrial Technological Development); Mr. Eduardo Díaz (Head of Unit New Technology Based Firms Unit, Fundación para el Cono-cimiento madri+d)
Moderator: Mr. Bonifacio Vega (Technology Transfer and Business Development Manager at IMDEA Nanociencia)
13:30 - 15:00 LUNCH BREAK
15:00 - 16:40 Poster session
Chairperson: Dr. Aitziber L. Cortajarena, IMDEA Nano-ciencia
16:40 - 17:10 Invited speaker: Dr Gray Kueberuwa, Institute of Cancer Sciences, University of Manchester
“Chimeric Antigen Receptor T-cells for Cancer Therapy”
17:10 - 17:30 Dr. Antonio Benayas, University of Quebec “At the Fluorescent Nanoprobes´ Frontiers: Multifunc-
tionality for Bio-Imaging and T-Sensing”
17:30 - 17:50 Dr. Ángel Millán, Instituto de Ciencias Materiales de Aragón
“Joining time-resolved thermometry and magnetic-induced heating in a single nanoparticle”
20:30 NETWORKING DINNER
Wednesday 25 February Session 3: NanoTherapy
Chairperson: Dr. Mª del Puerto Morales, Instituto de Ciencia de Materiales de Madrid-CSIC
09:30 - 10:15 Keynote speaker: Prof. Ingrid Hilger, University Hos-pital Jena
“Magnetic hyperthermia, a promising tool for the minimal-invasive treatment of tumors”
10:15 - 10:35 Dr. A. Roig, Instituto de Ciencia de Materiales de Bar-celona-CSIC
“Encapsulation of VEGF165 in magnetic PLGA nano-capsules for potential local delivery and bioactivity into human brain endothelial cells”
10:35 - 10:55 Mr. D. Cabrera, IMDEA Nanociencia “Influence of nanoparticle size and field frequency on
the concentration dependence of magnetic heating”
10:55 - 11:15 Ms. Blanca del Rosal, Universidad Autónoma de Madrid “Intratumoral thermal reading during photothermal
therapy by multifunctional fluorescent nanoparticles”
11:15 - 11:35 Prof. Nguyen TK Thanh, University College London “Magnetic nanoparticle-based therapeutic agents for
thermo-chemotherapy treatment of cancer”
11.35 -12:00 COFFEE BREAK
Chairperson: Dr. Sara Trabulo, Barts Cancer Institute
12:00 - 12:20 Dr. Cristina Fornaguera, Instituto de Química Avanzada de Cataluña-CSIC
“Nano-emulsion templating: a versatile technology to prepare multifunctional polymeric nanoparticles for advanced biomedical applications”
12:20 - 12.40 Dr. Anna Aviñó, Instituto de Química Avanzada de Cataluña-CSIC
“G-quadruplex aptamers with therapeutic applications”
12:40 - 13:30 Round table on nanotherapy: small treats different.
Members: Prof. Ingrid Hilger (University Hospital of Jena), Dr. Domingo F. Barber (Centro Nacional de Biotecnología-CSIC), Dr. Petra Gener (Hospital Vall d’Hebron)
Moderator: Dr. Aitziber L. Cortajarena, IMDEA Nano-ciencia
13:30-15:00 LUNCH BREAK
Chairperson: Dr. Pilar Calvo, Pharmamar
15:00 - 15:30 Invited Speaker: Dr. Petra Gener, Hospital Vall d’Hebron “Targeted Drug Delivery Systems against Cancer Stem
Cells”
15:30 - 15:50 Ms. Alicia Soler Cantón, Delft University of Technology “Development of gene-expressing liposomes as drug
delivery systems”
15:50 - 16:10 Dr. Laura Gallego-Yerga, Universidad de Sevilla “Self-assembling nanoparticles based on calixarene-
cyclodextrin heterodimers for targeted delivery and controlled release of docetaxel”
16:10 - 16:30 Mr. Gustavo da Silva, Instituto de Ciencia de Materiales de Madrid-CSIC
“Design of biocompatible magnetic nanoplatforms for potential platinum-based drug delivery applications”
16:30 - 17:00 CLOSING REMARKS
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ORAL contributions
Monday 23rd (morning session)
1. Nanodrugs Development and Applica-
tions to Pancreas Cancer, Dr. Manuel
Hidalgo, Centro Nacional de Investigaciones
Oncológicas
2. An overview of cancer and (drugabble-)
Immune response, Dr. José Antonio
López, Instituto de Investigación Hospital 12
de Octubre
3. Ageing and biotransformation of
nanomagnets in the body: How to con-
ciliate therapeutic efficiency and safe
life-cycle, Prof. Florence Gazeau, U. Paris
Diderot-CNRS
Monday 23rd (afternoon session)
4. Safety of Nanomaterials: Immunotoxicol-
ogy and interaction between Nanoma-
terials and human proteins, Prof. Africa
González Fernández, Instituto de Investiga-
ciones Biomédicas de Vigo (UVIGO)
5. Effect of Polyethylenimine-coated SPIO-
NS on macrophage activation and podo-
some dynamics, Dr. Vladimir Mulens, Centro Nacional de Biotecnología-CSIC
6. Tracking magnetic nanoparticles in
tissues by AC magnetic susceptibility:
Recent achievements and future chal-
lenges, Dr. Lucía Gutiérrez, Instituto de
Ciencia de Materiales de Madrid-CSIC
7. Soft X-ray tomography as quantitative
tool to decipher the interaction between
DMSA-SPION and MCF7 cancer cells,
Dr. Francisco Javier Chichón, Centro
Nacional de Biotecnología-CSIC
8. Multiparamagnetic High Throughput
Methods for Magnetic Iron Oxide Nano-
particle Lead Selection, Mr. K. Crosbie-
Staunton, Trinity College Dublin
Tuesday 24th (morning session)
9. Tracer development critical for transla-
tional medical applications of Magnetic
Particle Imaging, Prof. Kannan Krishnan, University of Washington
10. Early detection of Pancreatic Cancers &
Brain Tumors by MR Molecular Imaging,
Prof. Yung-Ya Lin, University California, Los
Angeles
11. Parallel multifuncionalisation of nano-
particles: A One-Step Modular Approach
for in vivo Imaging, Dr. Fernando Herranz, Centro Nacional de Investigaciones Cardio-
vasculares
12. Improving magnetic properties of ultras-
mall magnetic nanoparticles by biocom-
patible coatings, Dr. Rocío Costo, Instituto
de Ciencia de Materiales de Madrid-CSIC
13. Synthesis and characterization of bimetal-
lic magnetic oxides obtained by oxidative
precipitation, Ms. Yurena Luengo, Instituto
de Ciencia de Materiales de Madrid-CSIC
14. Coating strategies for the application of
magnetic nanoparticles in cancer diagno-
sis and therapy, Dr. Marzia Marciello, Insti-
tuto de Ciencia de Materiales de Madrid-CSIC
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15. Beyond the frontiers of the cell: unveiling
the wonders of nature at molecular level,
Dr. Wim Busing, FEI
Tuesday 24th (afternoon session)
16. Chimeric Antigen Receptor T-cells for
Cancer Therapy, Dr. Gray Kueberuwa, Institute of Cancer Sciences, University of
Manchester
17. At the Fluorescent Nanoprobes’ Fron-
tiers: Multifuncionality for Bio-Imaging
and T-sensing, Dr. Antonio Benayas, University of Quebec
18. Joining time-resolved thermometry and
magnetic-induced heating in a single
nanoparticle, Dr. Ángel Millán, Instituto
de Ciencias de Aragón
Wednesday 25th (morning session)
19. Magnetic hyperthermia, a promising
tool for the minimal-invasive treatment
of tumors, Prof. Ingrid Hilger, University
Hospital Jena
20. Encapsulation of VEGF165 in magnetic
PLGA nanocapsules for potential local
delivery and bioactivity into human brain
endothelial cells, Dr. A. Roig, Instituto de
Ciencia de Materiales de Barcelona-CSIC
21. Influence of nanoparticle size and field
frequency on the concentration depend-
ence of magnetic heating, Mr. David
Cabrera, Fundación IMDEA Nanociencia
22. Intratumoral thermal reading during
photothermal therapy by multifunctional
fluorescent nanoparticles, Ms. Blanca del
Rosal, Universidad Autónoma de Madrid
23. Magnetic nanoparticle-based therapeu-
tic agents for thermo-chemotherapy
treatment of cancer, Prof. Nguyen TK
Thanh, University College London
24. Nano-emulsion templating: a versatile
technology to prepare multifunctional
polymeric nanoparticles for advanced
biomedical applications, Dr. Cristina
Fornaguera, Instituto de Química Avanzada
de Cataluña-CSIC
25. G-quadruplex aptamers with therapeutic
applications, Dr. Anna Aviñó, Instituto de
Química Avanzada de Cataluña-CSIC
Wednesday 25th (afternoon session)
26. Targeted Drug Delivery Systems against
Cancer Stem Cells, Dr. Petra Gener, Hos-
pital Vall d’Hebron
27. Development of gene-expressing lipo-
somes as drug delivery systems, Ms.
Alicia Soler Cantón, Delft University of
Technology
28. Self-assembling nanoparticles based on
calixarene-cyclodextrin heterodimers for
targeted delivery and controlled release
of docetaxel, Dr. Laura Gallego Yerga, Universidad de Sevilla
29. Design of biocompatible magnetic nan-
oplatforms for potential platinum-based
drug delivery applications, Mr. Gustavo
da Silva, Instituto de Ciencia de Materiales
de Madrid-CSIC
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POSTER contributions
23rd and 24th afternoon session
P1. Interaction of multifunctional magnet-
ic nanoparticles with biological mem-
branes, Ms. María José Rodríguez, Centro Nacional de Biotecnología-CSIC
P2. Biodistribution of magnetic nano-
particles evaluated by AC magnetic
susceptibility in a murine model, Mr.
Eduardo Lorente-Sorolla, Instituto de
Ciencia de Materiales-CSIC
P3. C. elegans: an in vivo model to evaluate
nanoparticles?, Dr. A. Roig, Instituto de
Ciencia de Materiales de Barcelona-CSIC
P4. Iron quantification inside cells
incubated with SPION by Soft-Xray
Absorption Spectro-Tomography
(SXAST), Mr. José Javier Conesa, Centro Nacional de Biotecnología-CSIC
P5. Following the transformation of mag-
netic nanoparticles over time in mac-
rophages by AC magnetic susceptibil-
ity, Ms. Vanesa del Dedo, Instituto de
Ciencia de Materiales de Madrid-CSIC
P6. pH-Dependent Switch for Anticancer
Metallodrugs, Mr. Francisco Martínez, Fundación IMDEA Nanociencia
P7. Superparamagnetic properties of
hydrothermally prepared CoFe2O4
as a function of size (6–10 nm) and
coating (oleic/citric acid or TiO2), Dr.
Daniel Nižnanský, Charles University in
Prague
P8. Targeting cancer cells with photoac-
tive silica nanoparticles, Ms. Wioleta
Borzecka, University of Aveiro
P9. Combinational sensitization of tumor
cells with two photosensitizers syn-
ergistically enhances their photody-
namic inactivation in vitro, Ms. And-
rea Tabero, Universidad Autónoma de
Madrid
P10. Synthesis Strategies of Single-Core
Magnetic Nanoparticles, Ms. Helena
Gavilán, Instituto de Ciencia de Materi-
ales de Madrid-CSIC
P11. Silica encapsulation of magnetic nan-
oparticles, Ms. Leonor de la Cueva, Fundación IMDEA Nanociencia
P12. Synthesis of hybrid magneto-plas-
monic nanostructures based on Au
nanorods and iron oxides nanoparti-
cles, Mr. Jesús G. Ovejero, Universidad
Complutense de Madrid
P13. Functionalized magnetic nanoparti-
cles for cancer therapy, Dr. Antonio
Aires, Fundación IMDEA Nanociencia
P14. Detection and Inhibition of Mutated
GNAQ Gene using Spherical Nucleic
Acids, Ms. Ana Latorre, Fundación
IMDEA Nanociencia
ORAL contr ibut ion
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2 .An overview of cancer and (druggable-) immune responseDr. José Antonio LópezTranslational Oncology Group co-Director,`12 de Octubre’ Research Institute Attending Physician,
‘12 de Octubre’ University Hospital
Abstract: The immune system has a great potential for the specific destruction of tumours
and for long-term memory that can prevent cancer recurrence. The probability of an effec-
tive tumour immune response depends on 1- the existence of tumour antigens and 2- the
possibility of modulating the mechanisms of suppression of tumour immune response.
Tumour escape from immunosurveillance be achieved either by changes in the tumour cells
(loss of tumour antigens, loss of human leukocyte antigen molecules, loss of sensitivity to
complement, or T cell or natural killer (NK) cell lysis) or by tumour subvertion of the normal
immune regulation. Tumour immunity can be enhanced by blocking inhibitory pathways and
inhibitory cells in the tumour microenvironment, or by increasing the specificity of antitumour
immunity, inducing the expansion of T cells and antibodies directed to well-defined tumour
antigens (e.g. cancer vaccines, potent adjuvants, immunostimulatory cytokines). These
strategies can be pursued either alone or as part of a multimodal therapy.
3 .Ageing and biotransformation of nanomagnets in the body: How to conciliate therapeutic efficiency and safe life-cycle?Florence GazeauMatière et Systèmes Complexes, CNRS / Université Paris Diderot
75205 Paris Cedex 13, FRANCE
The increasing exposure of living organism to nanoparticles calls for a comprehensive assess-
ment of the processing of nanomaterials in vivo. Understanding nanoparticles distribution,
transformations, persistence, ageing and degradation will help to predict exposure risks as
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well as therapeutic efficiency of nanoparticles. The challenge is to establish how the synthetic
identity of nanoparticles can determine their whole life cycle in the body.
We have developped a multiscale methodology to examine the influence of biological envi-
ronment on the structure and physical properties of magnetic nanoparticles that show
outstanding properties for magnetothermal therapy and MRI detection.
Our approach, which is based on materials sciences, is combining nanoscale electron micro-
scopy observations of nanoparticle with the follow-up of magnetic properties in biological
environments. From several examples of magnetic nanostructures – iron oxide nanospheres,
nanocubes, cooperative nanoflowers and iron oxide/gold dimers with different coating – we
will examine how cell-induced transformations critically alter the nanoparticles magnetic
properties, heating power and Magnetic Resonance relaxivity over time and determine the
long term fate of the nanoparticles in the organism. In the research for safe and efficient
nanoparticles for nanomedecine, one should control the balance between short term efficacy
in the relevant biological context and long term degradability and clearance from the body.
References1. Lartigue L, Alloyeau D, Kolosnjaj-Tabi J, Javed Y, Guardia P, Riedinger A, Péchoux C, Pellegrino T, Wilhelm C,
Gazeau F. Biodegradation of Iron Oxide Nanocubes: High-Resolution In Situ Monitoring. ACS Nano 2013, 7, 3939-3952.
2. Kolosnjaj-Tabi J, Di Corato R, Lartigue L, Marangon I, Guardia P, Silva AK, Luciani N, Flaud P, Clément O, Singh JV, Decuzzi P, Pellegrino T, Wilhelm C, Gazeau F. Heat-Generating Iron Oxide Nanocubes: Subtle “Destructu-rators” of the Tumoral Microenvironment. ACS Nano 2014, May 27;8(5):4268-83
3. Javed Y, Lartigue L,Hugounenq P, Vuong QL, Gossuin Y, Bazzi R, Wilhelm C, Ricolleau C, Gazeau F*, Alloyeau A*. Biodegradation mechanism of iron oxide monocrystalline nanoflowers and tunable shield effect of gold coating, Small, 2014 Aug 27;10(16):3325-37
4 .Safety of nanomaterials: Immunotoxicology and interaction between Nanomaterials and human proteins. Prof. África González-FernándezMD, PhD, Immunologist
Professor of Immunology
Director of the Biomedical Research Center (CINBIO)
Coordinator of BIOCAPS (EU 7º FW program)
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UNIVERSITY OF VIGO Campus Lagoas Marcosende
36310 Vigo, Pontevedra
Spain
phone: +34-986812625
fax: +34-986812556
email: [email protected]; [email protected]
The use of nanomaterials (NMs) is increasing in the last years because of their potential to
improve a wide range of applications, especially in the biomedical field. However, NMs can
interact with the body inducing undesirable or desirable effects. It is crucial to understand
their potential harmful effects, and the success of the NMs on biomedical applications will
partially depend on their interactions with blood and immune components. The immune
system has the role, between other functions, of defending the body from foreign materials.
The study of the interaction between the components of the immune system and NMs is,
therefore, an area of great interest. Moreover, NMs can be used in biomedicine to target
different cells to modulate their function towards a desired response, for example in vac-
cines. The talk will focus on how the interactions between NMs and biological systems could
affect to the normal function of these systems.
5 .Effect of Polyethylenimine-coated SPIONs on macrophage activation and podosome dynamicsAuthors: Vladimir Mulens1, José Manuel Rojas1, Sonia Pérez-Yagüe1, María del Puerto Mora-
les2, Domingo F. Barber1*
1. Department of Immunology and Oncology and NanoBiomedicine Initiative, Centro Nacional de
Biotecnología (CNB)/CSIC, Darwin 3, Cantoblanco, 28049 Madrid, Spain
2. Department of Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid
(ICMM)/CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
Mulens V. email address: [email protected]
Huge efforts have been made to develop multifunctional platforms for cancer gene therapy.
One of the current strategies is the combination of SPIONs (superparamagnetic iron oxide
nanoparticles) and polyethylenimine (PEI), due to their unique chemical and physical proper-
ties. PEI appears to activate different immune cells toward an inflammatory response (M1/
TH1), whereas the response induced by SPIONs seems to be controversial. Nonetheless,
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the immunoreactivity of PEI-coated SPIONs has not been addressed so far. In the present
study, we proved that PEI-coated SPIONs (PMag) induce an M1 phenotype. PMag induced
phosphorylation of p38 MAPK, p44/p42 MAPK and JNK, and upregulation of CD40, CD80,
CD86 and I-A/I-E activation markers. Importantly, PMag-induced macrophage activation
depended partially on TLR4 (Toll-like receptor 4) and ROS (reactive oxygen species) signal-
ing, and was different from that induced by LPS. Intriguingly, PMag promoted podosome
formation in murine macrophages, but inhibited gelatin degradation. This effect seems to
be due to the overexpression of genes related with MMP inhibitors and downmodulation
of active MMP-2. In conclusion, PMag induced an M1-like phenotype that was partially
dependent on both TLR4 and ROS, and modulated the dynamics of podosomes.
6 .Tracking magnetic nanoparticles in tissues by AC magnetic susceptibility: Recent achievements and future challengesLucía Gutiérreza, Raquel Mejíasb,c, Domingo F. Barberb, Francisco J. Lázarod and M. Puerto
Moralesa
a. Dept. Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid (ICMM)/
CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
b. Dept. Immunology and Oncology, and NanoBiomedicine Initiative, Centro Nacional de Biotecnología
(CNB)/CSIC, Darwin 3, Cantoblanco, 28049 Madrid, Spain
c. Department of Biomedical Engineering, Vanderbilt University, 5824 Stevenson Center, Nashville,
Tennessee 37235-1631, USA
d. Dept. de Ciencia y Tecnología de Materiales y Fluidos, Universidad de Zaragoza, Maria de Luna
3, 50018, Zaragoza, Spain
The quantitative determination of magnetic nanoparticles in tissue samples from animal
models using magnetic measurements is helpful but not free of difficulties. Once the particles
enter the body, they become surrounded by a wide variety of proteins and other molecules,
altering their environment, possibly their degree of aggregation, and therefore the magnetic
interparticle interactions. Also, due to ultrastructural differences, each organ may present
a characteristic way of packing the particles forfurther processing that can also result in
an alteration of the magnetic properties of the material. Furthermore, these systems suffer
a continuous transformation process, where degradation of the particles also affects the
magnetic properties of the whole tissue. Due to these complex factors, the comparison
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between the magnetic properties of the administered compound and those of the tissue
samples containing the particles is not always straightforward.
Based on AC magnetic susceptibility measurements of freeze-dried tissue samples, we
have developed a protocol to quantify magnetic nanoparticles in animal organs [1]. This
characterization protocol has been recently used to study the biodistribution of magnetic
nanoparticles in mouse models, and the effects of the particle coating on the biodistribution
[2,3]. It has also been used to follow particle transformations over long time periods [4]. The
advantages of this technique include the possibility to distinguish the particles from other
endogenous species such as the ferritin iron cores, and that no prior isolation procedures,
potentially altering the results, are required. Nevertheless, especial attention has to be paid
to the influence of interparticle dipolar interactions in the different tissues and the possible
reduction of the particle size due to degradation processes, as both parameters affect the
magnetic susceptibility results. We will describe the developed quantification protocol and
our recent results on the quantification of magnetic nanoparticles in tissue samples. Future
possible approaches to improve this characterization technique will be discussed.
Figure 1. Schematic representation of some parameters that affect the out-of-phase susceptibil-
ity used for the quantitative determination of magnetic nanoparticles in tissue samples.
References1. L. Gutiérrez et al. Phys.Chem.Chem.Phys.,16 (2014) 4456.2. R. Mejías et al. Biomaterials, 32 (2011) 2938.3. L. Gutiérrez et al. J. Phys. D, App, Phys.,44 (2011) 2550024. R. Mejías et al. J. Controlled Release, 171 (2013) 225.
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7 .Soft X-ray tomography as quantitative tool to decipher the interaction between DMSA-SPION and MCF7 cancer cellsFrancisco Javier Chichóna, José Javier Conesa1a, Michele Chiappia, Eva Pereirob, Maria Josefa
Rodrigueza, and José L. Carrascosaa,c
a. Centro Nacional de Biotecnología (CNB-CSIC), Cantoblanco, 28049 Madrid, Spain.
b. ALBA Synchrotron Light Source. MISTRAL Beamline - Experiments division. 08290 Cerdanyola
del Vallès, Barcelona, Spain.
c. Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), Cantoblanco,
28049 Madrid, Spain
In the last years the use of nanotechnology in medicine opened a broad field for studying cell-
nanoparticle interaction. Due to their relatively easy generation and their low toxicity, iron magnetic
nanoparticles are nowadays very promising. Superparamagnetic nanoparticles (SPION) could be
coated with specific agent to target cells for drug delivery. Their magnetic properties also allow the
manipulation of threated cells for cellular sorting or to induce cellular death by processes, such as
hyperthermia. Those are feasible and important objectives in nanobiotechnology. In this frame, we
have used Soft-X-ray microscopy to decipher and quantify the interaction of superparamagnetic
iron oxide nanoparticles with average diameter of 15 nm (DMSA-SPION) with MCF-7 human
breast cancer cells [1]. SPION were incubated from 0 to 24 h with cells and cryo-preserved
by plunge freezing. We also performed correlative microscopy to locate by cryo-epifluorescent
microscopy the Internalized SPION accumulations in the endocytic pathway and therefore, the
samples were submitted to cryo Soft X-ray tomography (cSXT) to get whole cell volume maps.
This is an emerging three-dimensional technique that exploits Soft X-rays high penetration into
the biological matter to imaging vitrified cells at nanometric 3D resolution without chemical fixa-
tion or staining agents [2]. The three-dimensional reconstructions of whole cells allowed us to
perform statistical analysis of SPION accumulation. Our results enlighten parameters such the
average vesicles size, distance between them, distance to the nucleus and accumulation rates
that characterize this type of accumulation. This new approach opens a new door to study in a
quantitative way and with high resolution the interaction of nanoparticles and cells.
References1. [1] M.Calero, M.Chiappi, et al., “Characterization of interaction of magnetic nanoparticles with breast cancer
cells”. Journal: Journal of Nanobiotechnology “in press”.
2. [2] Pereiro, Eva; and Chichón, Francisco Javier (November 2014) Cryo-Soft X-ray Tomography of the cell. In: eLS. John Wiley & Sons, Ltd: Chichester. DOI: 10.1002/9780470015902.a0025582
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8 .Multiparametric High Throughput Methods for Magnetic Iron Oxide Nanoparticle Lead Selection
K. Crosbie-Staunton1*, G. Salas2,3, O. Gobbo4, M. Marciello2,3, P. Couleaud3, B. M Mohamed1,
T. Rakovich1, R. Miranda5,6, M. del Puerto Morales2, A. Prina-Mello1,7, Y. Volkov1,7
1. School of Medicine, Trinity College Dublin, Ireland.
2. Instituto de Ciencia de Materiales de Madrid/CSIC, Madrid, Spain.
3. IMDEA Nanociencia, Madrid, Spain.
4. School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Ireland.
5. Instituto Madrileño de Estudios Avanzados, IMDEA Nanociencia, Madrid, Spain.
6. Departamento de Física de la Materia Condensada, Universidad Autónoma de Madrid, Spain.
7. CRANN and AMBER Research Centres, Trinity College Dublin, Ireland.
* Corresponding author, [email protected]
Introduction: The application of magnetic nanoparticles (MNPs) can be tailored on a case
by case basis by coupling them simultaneously with targeting moieties and medicinal drugs,
which make multifunctional (MF) MNPs suitable for theranostic applications. Their poten-
tial role as magnetic resonance imaging (MRI) contrast agents, in drug and gene delivery
platforms and hyperthermia are being intensely researched [4]. This study, as part of the
MULTIFUN FP7 project, focuses on the multiparametric biocompatibility testing of a range
of MNPs to select lead formulations. Further testing of the selected leads following function-
alization with a targeting ligand (NUCANT 6L, N6L) and chemotherapeutic drugs (Doxoru-
bicin and Gemcitabine) in breast cell lines was conducted to identify suitable theranostic
MF-MNPs. Experimental procedure: Physicochemical characterisation of MNPs was carried
out by transmission electron microscopy, dynamic light scattering and nanoparticle tracking
analysis prior to cell exposure. A high throughput approach to determine biocompatibility
was implemented with a range of MNP concentrations tested in breast-derived cell lines.
Multiparametric analysis was conducted to determine the changes in the total cell count,
cell membrane permeability and lysosomal mass/pH while flow cytometry was employed
to further investigate the effect on cell cycle progression with one lead MNP. Main results:
OD15 and MF66 MNPs below 100 µg/mL did not reduce cell viability or induce damage to
the cell membrane, and were selected as the lead formulations for functionalization. Cell
cycle analysis by flow cytometry with cells exposed to a range of MF66 MNP concentra-
tions was conducted to ensure that MF66 was suitable to bring forward. It was determined
that MF66 did not alter the normal cell cycle at concentrations below 100 ug/mL and was
considered a suitable lead formulation. N6L was previously shown to target nucleolin and
nuclophosmin on the cell surface and inhibit malignant tumour growth, angiogenesis and
cancer cells invasion alike [5]. We demonstrate that free N6L exposure at 72 h timepoint
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reduces cell count in a dose dependent manner and is most pronounced in breast cancer
cell lines compared to the normal-like breast-derived cell line, which is consistent with previ-
ously published data. However, functionalised OD15 and MF66 MNPs containing N6L did
not demonstrate similar efficacy compared to the free ligand. Therefore, we utilised N6L as
a targeting moiety to facilitate greater MNP uptake into the MCF-7 and MCF-10A cells with
chemotherapy drug-functionalised MNPs. We found that N6L-functionalised MF-MNPs
had enhanced uptake and chemotherapeutic drug delivery efficacy compared to N6L-free
MF-MNPs. Conclusion: This study demonstrates that by implementing an in-depth investiga-
tion of physicochemical properties and multiparametric cytotoxicity analysis, it is possible
to select MNP formulations and subsequently select targeted MF-MNP leads for transfer to
in vivo animal models and clinical exploitation.
Acknowledgement: This work is partially funded by the European Commission under the
FP7 programme (MULTIFUN project: NMP4-LA-2011-262943)
References:1. Gultepe E, Reynoso FJ, Jhaveri A et al.: Monitoring of magnetic targeting to tumor vasculature through MRI and
biodistribution. Nanomedicine (Lond) 5(8), 1173-1182 (2010).
2. Roca AG, Veintemillas-Verdaguer S, Port M, Robic C, Serna CJ, Morales MP: Effect of nanoparticle and aggregate size on the relaxometric properties of MR contrast agents based on high quality magnetite nanoparticles. The journal of physical chemistry. B 113(19), 7033-7039 (2009).
3. Corr SA, O’byrne A, Gun’ko YK et al.: Magnetic-fluorescent nanocomposites for biomedical multitasking. Chemical communications (43), 4474-4476 (2006).
4. De Jong WH, Borm PJ: Drug delivery and nanoparticles:applications and hazards. International journal of nano-medicine 3(2), 133-149 (2008).
5. Destouches D, Huet E, Sader M et al.: Multivalent pseudopeptides targeting cell surface nucleoproteins inhibit cancer cell invasion through tissue inhibitor of metalloproteinases 3 (TIMP-3) release. The Journal of biological chemistry 287(52), 43685-43693 (2012).
9 .Tracer development critical for translational medical applications of Magnetic Particle Imaging
Kannan M. KrishnanDepartment of Materials Science & Engineering
University of Washington, 323 Roberts Hall,
Box 352120, Seattle, WA 98195
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The performance of Magnetic Particle Imaging (MPI), a new whole-body medical imaging
technology with promise in vascular angiography and molecular imaging, depends largely
on the structural, chemical, magnetic, and biological characteristics of the tracers. In fact,
MPI is the first biomedical imaging technique that truly depends on nanoscale materials
properties; in particular, their response to alternating magnetic fields in a true biological
environment needs to be optimized.
I will describe the development of our highly optimized nanoparticle tracers with magnetic prop-
erties tailored for the unique physics of MPI using a sound theoretical framework, an organic
synthetic route producing phase-pure magnetite cores with controlled shape and narrow
size-distribution. Their subsequent phase-transfer without agglomeration, even in biologically
relevant media, and functionalization for targeting, biocompatibility, adequate in vivo circulation
(tailorable between 4-80 minutes in mice models) and tailorable biodistribution (no uptake
in the kidneys, critical for imaging patients with chronic kidney disease) will be presented.
In MPI phantom images, reconstructed in either real or Fourier space, these optimized trac-
ers showed significant improvement compared to existing commercial particles (Resovist®),
in both normalized signal intensity (6x) and spatial resolution (50% better) approaching sub-
mm at 6 T/m/µ0 field gradients. Further, their MPI signal is linear with concentration, with
minimal contribution from Brownian relaxation, making them suitable for molecular imaging.
Finally, we have established a flexible platform for functionalization of the magnetic cores
with PEG (to increase water solubility, enhance circulation time, and improve shelf life),
dyes (to turn them into multimodal -- MRI, MPI and NIRF -- contrast agents and used to
further enhance studies of their biodistribution with significant anatomical detail in rodent
models) and ligands (such as Lactoferrin with specific affinity for glioma cells) tailored for
targeted molecular imaging.
These critical results, that pave the way for improved clinically relevant MPI performance,
will be discussed with emphasis on the tracer response to both the alternating magnetic
field and the biological environment.
This work was supported by NIH grants 1RO1EB013689-01 (National Institute of Biomedical
Imaging and Bioengineering, NIBIB) and 1R41EB013520-01.
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10 .Early Detection of Pancreatic Cancers & Brain Tumors by MR Molecular ImagingYung-Ya Lina, Zhao Lia, and Chaohsiung Hsua
a. Department of Chemistry and Biochemistry, University of California, Los Angeles
Introduction: Early detection of high-grade malignancy, such as pancreatic cancers (PC) and
glioblastoma multiforme (GBM), using enhanced MRI techniques significantly increases not
only the treatment options available, but also the patients’ survival rate. For this purpose,
a conceptually new approach, termed “Active-Feedback Controlled MR”, was developed.
An active feedback electronic device was homebuilt to implement active-feedback pulse
sequences to generate avalanching spin amplification, which enhances the weak field
perturbations from magnetic nanoparticles in targeted PC or malignant physiological condi-
tions in GBM.
Theory and Methods: The general principles of the “Active-Feedback Controlled MR” can
be found in our publications [1-6] (and references therein). Here, its specific applications
to early tumor detection were developed and demonstrated. (i) First, an active-feedback
electronic device was home-built to generate feedback fields from the received FID current.
The device is to filter, phase shift, and amplify the signal from the receiver coils and then
retransmit the modified signal into the RF transmission coil, with adjustable and programma-
ble feedback phases and gains. The MR console computer can execute the active-feedback
pulse sequences to control the trigger signal, feedback phase/gain, and the duration of the
feedback fields, allowing us to utilize the active feedback fields in novel ways. (ii) Next, an
active-feedback pulse sequence was developed for early tumor detection and was statisti-
cally tested on in vivo mice tumor models. In essence, the enhanced tumor contrast arises
from “selective self-excitation” and “fixed-point dynamics” generated by the bulk water 1H
under active feedback fields. Use the sensitive detection of magnetic nanoparticles as an
example. A small flip-angle (q=5-10o) RF pulse tilts the sample equilibrium magnetization.
Since the averaged transverse magnetization is mainly contributed from the bulk water 1H
spins, the resulting active feedback field possesses a frequency closer to that of the bulk
water 1H spins which are distant from the dipole center. By “selective self-excitation”, the
feedback field tilts the bulk water 1H spins more effectively towards the stable fixed-point,
–z-axis (assume feedback phase 180o), while the 1H spins near the dipole center are less
affected due to resonance mismatch. This “selective self-excitation” process continues and
enlarges the contrast between the longitudinal magnetization of the 1H spins in bulk water
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and those near the dipole center. Maximum contrast in the longitudinal magnetization can
be achieved and locked when all spin magnetizations evolve to the fixed point: all align
along –z in this case.
Results (1): Early Detection of Pancreatic Cancers: Anti-CA 19-9 antibodies were conjugated
to NH2-PEG-coated magnetic nanoparticles. The antigen binding capacity to CA 19-9 over-
expressing cell lines (BxPC3) was confirmed by in vitro MR cellular images. In vivo images of
human pancreatic tumors from nude mouse xenografts show that, while T2-weighted image
cannot clearly locate the magnetic nanoparticles, the active-feedback images successfully
highlight the magnetic nanoparticles distribution with a close correlation with iron-stained
histopathology.
Results (2): Early Detection of Glioblastoma Multiforme: Stage-1 orthotopic GBM mouse
models infected with human U87 cell line were imaged. While both T2 parameter images
and T2-weighted images by spin echo successfully locate the GBM tumor, our active-
feedback images and decay constant mapping provide 4-5 times of improvements in GBM
tumor contrast through sensitively imaging the susceptibility variations due to irregular water
contents and deoxyhemoglobin.
Discussion and Conclusion: In vivo PC and GBM mouse models validated the superior
contrast/sensitivity and robustness of the “Active-Feedback MR” for early tumor detection.
Statistical results (N>10) for PC and GBM mouse models at various cancer stages, alternative
active feedback pulse sequences with further improved performance, and active feedback
pulse sequences for enhanced R1/R2-weighted images will also be presented.
References1. Science 290, 118 (2001) 2. Magn. Reson. Med. 56, 776 (2006) 3. Magn. Reson. Med. 61, 925 (2009) 4. J. Phys. Chem. B 110, 22071 (2006) 5. Magn. Reson. Med. (2015, in press) 6. Biomaterials 32, 2160 (2011)
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11 .Parallel Multifunctionalisation of Nanoparticles: A One-Step Modular Approach for in vivo Imaging Fernando Herranza, Hugo Groulta, Juan Pellicoa, Ana V. Lechuga-Viecoa, Riju Bhavesha, Jesús
Ruiz-Cabelloa
a. Advanced Imaging Unit, Department of Atherotrombosis and Imaging, Fundación Centro Nacional
de Investigaciones Cardiovasculares (CNIC) and CIBER de Enfermedades Respiratorias (CIBERES),
Melchor Fernández-Almagro, 3, 28029 Madrid, Spain
Multifunctional nanoparticles are usually produced by sequential synthesis, with long
multistep protocols.1 Until now additional functionality meant additional synthetic steps
and costs, lower yields and a complex mixture on the surface of the particles.2 Our study
reports a generic modular strategy for the parallel one-step multifunctionalisation of differ-
ent hydrophobic nanoparticles.3 The aim was to apply the concept of parallel synthesis,
developed in organic chemistry, for the synthesis of multifunctional inorganic nanopar-
ticles. The method was designed and developed by taking advantage of the natural non-
covalent interactions between the fatty acid binding sites of the bovine serum albumin and
the aliphatic surfactants on different inorganic nanomaterials. As a general example of the
approach, three different nanoparticles – iron oxide, upconverting nanophosphors and gold
nanospheres – were nanoemulsified in water with albumin. To support specific applications,
multifunctional capability was incorporated with a variety of previously modified albumin
modules. These modules include different conjugated groups, such as chelating agents
for 68Ga or 89Zr and ligand molecules for enhanced in vivo targeting. A large library of 13
multimodal contrast agents was developed with this convergent strategy. This platform
allows a highly versatile and easy tailoring option for efficient incorporation of functional
groups. Finally, as demonstration of this versatility, a bimodal probe (PET/MRI) including
a maleimide-conjugated BSA was selectively synthesized with an RGD peptide for in vivo
imaging detection of tumour angiogenesis.
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Figure 1. General scheme for the parallel functionalisation of hydrophobic nanoparticles with ap-
plications in cell labelling, PET imaging (68Ga or 89Zr) and MRI (T1 and T2 contrast).
References1. F. Herranz, J. Ruiz-Cabello et al. Contrast Media Mol. Imaging 7(2012) 435.2. Z. Cheng, A Tsourkas et al. Science 338(2012) 903.3. H. Groult, F. Herranz et al. Bioconjugate Chem. (2015) doi: 10.1021/bc500536y
12 .Improving magnetic properties of ultrasmall magnetic nanoparticles by biocompatible coatingsRocio Costo,a M. Puerto Morales,a and Sabino Veintemillas-Verdaguera
a. Instituto de Ciencia de Materiales de Madrid (CSIC). Sor Juana Ines de la Cruz, 3. 28049 Madrid
During last decades much has been written about the use of magnetic iron oxide nanoparti-
cles for applications in biomedicine as contrast agents for magnetic resonance imaging, drug
delivery or hyperthermia cancer treatment [1]. In any case, particles should be coated with
biocompatible molecules to avoid aggregation, improve the colloidal stability and guarantee
the biocompatibility of the final product. However, as a result of the interactions between the
particle surface and the coating, magnetic properties can be modified [2]. Understanding
these changes in magnetic behavior is critical for developing magnetic nanoparticles for
biomedical applications since the properties of the final product may be greatly different
from those of the initial iron oxide cores.
To study this phenomenon, ultrasmall particles were synthesized by laser pyrolysis and fully
oxidized to maghemite by acid treatment. The surface of the magnetic nanoparticles was
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systematically coated with either phosphonate (phosphonoacetic acid or pamidronic acid)
or carboxylate-based (carboxymethyl dextran) molecules and the binding to the nanoparticle
surface was analyzed. Magnetic properties at low temperature show a decrease in coercivity
and an increase in magnetization after the coating process. Hysteresis loop displacement
after field cooling is significantly reduced by the coating, in particular for particles coated
with pamidronic acid, which show a 10% reduction of the displacement of the loop. We
conclude that the chemical coordination of carboxylates and phosphonates reduces the sur-
face disorder and enhances the magnetic properties of ultrasmall maghemite nanoparticles.
Figure 1. Left: Magnetization curves at 5 K of the coated and un-coated particles. Right: Colloidal
stabilization of the particles with the three studied coatings: phosphonoacetic acid (red dots), pamid-
ronic acid (blue squares) and carboxymethyl-dextran (green diamonds)
References
1. S. Laurent et al. Chem. Rev. 108 (2008) 2064 2. C.E. Vestal et al. J. Amer. Chem. Soc. 125 (2003) 9828
13 . Synthesis and characterization of bimetallic magnetic oxides obtained by oxidative precipitationY. Luengoa, M.A. Roldánb, M. P. Moralesa, M. Varelab,c and S. Veintemillas-Verdaguera
a. Instituto de Ciencia de Materiales de Madrid, ICMM-CSIC, Sor Juana Inés de la Cruz 3, Canto-
blanco, Madrid, Spain
b. Universidad Complutense de Madrid, Departamento de Fisica Aplicada III, Madrid 28040, Spain
c. Materials Science and Technology Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831,
United States
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Bimetallic magnetic oxides in nanoparticulated form have plenty of potential applications
in many fields such as biomedicine (MRI contrast agents, magnetic hyperthermia agents,
drug delivery, biosensors), technology (magnetic recording), environmental remediation
(removal of pollutants with magnetic nanoparticles), etc. due to the combination of proper-
ties of both oxides. In the biomedical field, iron oxide is the most usual material because of
its low toxicity, high biocompatibility and good magnetic properties. However, there is still
plenty of room for research; for instance special features could be obtained by combination
of iron oxide nanoparticles with another metal oxide in a single entity.
Thus, bimetallic nanoparticles consisting of magnetite and other metal oxide, Fe3O4@MOx
(M: Bi, Gd, Co), were obtained by oxidative precipitation in aqueous medium. The different
metals were chosen due to their special properties: the presence of Bi may confer TC con-
trast while Gd could be used as T1 contrast agent in MRI and Co will enhance the magnetic
properties of the iron oxide particles. The maximum amount of metal oxide incorporation
without segregation and the type of distribution (solid solution, core-shell, decoration with
oxide particles, etc) were studied by scanning transmission electron microscopy (STEM)
and electron energy loss spectroscopy (EELS). The materials were characterised by X-Ray
diffraction, thermogravimetric analysis, infrared spectroscopy and chemical analysis. The
saturation magnetization, initial susceptibility and coercivity of all the samples were meas-
ured at room temperature and 5 K to study the effect of these metal oxides on the magnetic
properties of magnetite.
Figure 1. HAADF STEM micrographs of nanoparticles doped with: a) Bi, b) Gd and c) Co
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14 .Coating strategies for the application of magnetic nanoparticles in cancer diagnosis and therapy Marzia Marciello,a Marlène Branca,b,c Susana I.C.J. Palma,d Maryam Zahraei,e Diana Ciucu-
lescu-Pradines,b,c Behshid Behdadfara,e Ana Cecilia A. Roque,d Catherine Amiensb,c and M.
Puerto Moralesa
a. Biomaterials and Bioinspired Materials Dpt., Instituto de Ciencia de Materiales de Madrid (ICMM-
CSIC), Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain;
b. CNRS, LCC (Laboratoire de Chimie de Coordination), 205, route de Narbonne, F-31077 Toulouse,
France;
c. Université de Toulouse, UPS, INPT; LCC; F-31077 Toulouse, France;
d. UCIBIO, REQUIMTE, Chemistry Dpt., Universidade Nova de Lisboa, 2829-516 Caparica, Portugal;
e. Materials Engineering Dpt., Isfahan University of Technology, Isfahan 84156-83111, Iran.
In the last decades the use of magnetic nanoparticles (MNPs) for cancer diagnosis and
therapy has quickly increased. An important requirement for MNPs in vivo application
is the presence of an opportune coating that stabilizes them in physiological conditions,
and provides multifunctionality, biocompatibility and protection of magnetic core against
degradation.
In this work MNPs synthesized by different routes (aqueous and organic medium) [1-4]
have been coated using some opportune strategies: ligand exchange [2,3,4] superficial
chemical modification [1,3,4] and encapsulation [4] (Scheme 1).
Small molecules (DMSA, citric acid, TEOS, APTS) or biocompatible polymers (dextran,
polyethylene glycol, chitosan, gum arabic) have been strongly attached to MNPs surface
to avoid coating loss in physiological conditions. The objective was to make these particles
suitable as contrast agents for MRI or therapeutics for hyperthermia. All the coated MNPs
were characterized by good stability in physiological conditions and displayed opportune
size for biomedical application.
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Scheme 1: Different coating strategies used to enable MNPs bioapplication: A) Ligand exchange,
B) Chemical modification and C) Encapsulation.
References1. Marciello M. et al., J. Mat. Chem. B., 1 (2013) 5995.2. Kossatz S. et al. Pharm. Res., 31 (2014) 3274.3. Palma S. et al., J. Colloid Interf. Sci., 437 (2015) 147.4. Branca M. et al., J. Magn. Magn. Mater., 377, (2015) 348.
15 .Chimeric Antigen Receptor T-cells for Cancer TherapyDr David Gilham Institute of Cancer Science - University of Manchester, Manchester, UK
One mechanism employed by tumors to avoid immune medicated destruction is reduced or
loss of expression of Major Histocompatibility Complex (MHC) proteins which are essential
for T cell recognition processes. To overcome this avoidance strategy, CAR technology has
been developed that exploits antibody-based target binding thereby allowing the T-cell to
recognise cell surface tumor associated antigen directly thereby circumventing loss of MHC
expression. The CAR consists of four key elements; firstly, the antigen binding domain that
is most commonly a single chain antibody fragment (scFv) derived from an antibody but can
be derived from any potential ligand. Secondly, an extracellular spacer domain that allows
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for optimal binding of the scFv to its cognate target. Thirdly, a transmembrane domain that
tethers the CAR to the surface of the T cells and finally, an intracellular signalling domain
that engages with downstream pathways that initiate T-cell effector function upon ligation
of the CAR with target antigen. However, to date there has been consensus single optimal
configuration. Consequently, many different CARs are currently in clinical testing. These
will be discussed as well as recent developments in the CAR T cell field.
17 .At the Fluorescent Nanoprobes´ Frontiers: Multifunctionality for Bio-Imaging and T-SensingAntonio Benayas,a Fuqiang Rena, Blanca del Rosalb,Wagner F. da Silvac, Elisa Carrascob,
Francisco Sanz-Rodríguezb, Vicente Marzal2, Karla Santacruz-Gómezd, Gustavo A. Hiratae,
Dongling Maa, Daniel Jaqueb, Fiorenzo Vetronea.
a. Univ Quebec, INRS-EMT, 1650 Blvd. Lionel Boulet, Varennes, PQ J3X 1S2, Canada.
b. UAM, Cantoblanco 28049, Madrid, Spain.
c. UFAL, Maceió, 57072-900 Alagoas, Brazil.
d. Univ Sonora, Hermosillo 83000, Sonora, Mexico.
e. CNyN, Ensenada, 22800 Baja California, Mexico
Nowadays, new and better light-emitting probes are being demanded to fulfill the needs
and specific requirements of nanomedicine, especially at the cancer diagnosis and therapy
battleground. There is an intense appetite for multifunctional, non-invasive and biocompat-
ible platforms to be designed and implemented.
In this work on fluorescent nanoparticles (NPs) for bio-imaging, we present our last outstand-
ing results obtained throughout a research lines promoted at Vetrone group (INRS-EMT,
Canada). Starting with dielectric hosts, a remarkable outcome has been reached using
NaGdF4 lanthanide-doped NPs for cell imaging. They provide an extended colour-palette
within both visible and near-infrared spectral ranges, using two alternative excitation wave-
lengths (793 and 980nm). It should also be highlighted the pivotal role that fluorescent
nanothermometry has been playing within our investigations. Thus, the second project here
explained is constituted by the development of efficient Nd:YAG near infrared fluorescent
nanothermometers1,3, that have demonstrated remarkable performance on a broad range
of temperature-monitoring applications (electrical circuits, sub-tissue measurements and
optical trapping context). Finally, our most standing platform PbS/CdS quantum dots (QDs)
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for in vivo imaging, which wavelength-wise design (they emit ~ 1.3µm) increases penetration
depth2, and their high-brightness allows a remarkable low dose of them to provide fluores-
cence images, upon injection in live animals. This system has also shown some promissory
nanothermometry features, demonstrated through ex-vivo experiments.
References1. D. Jaque et al. Nanoscale 4 (2012) 4301-4326.2. T. Lim et al Mol. Imaging 2 (2003) 50-64.3. A. Benayas et al. Advanced Optical Materials (2015) DOI: 10.1002/adom.201400484
Figure 1: a) Left: graph showing the evolution of the QDs emitted intensity from three different
parts of the mouse body (tail/green, liver/red and lungs/blue) as a function of time after injection
inside the mouse´s body. Right: Combination of optical and fluorescence images taken from the
live specimen. b) Left: Schematic representation of an optical trapping setup. A 1480 nm laser
is focused into a microchannel filled with Nd:YAG NPs. Right: Thermal image of a 900 µmx900
µm area around the laser focus. Laser power was set to 62 mW. Figure taken from the author’s
recently published Ref. [3].
18 .Joining time-resolved thermometry and magnetic-induced heating in a single nanoparticleAngel Millan,a Rafael Piñol,a Carlos D. S. Brites,b Rodney Bustamante,a Abelardo Martínez,c
Nuno J. O. Silva,b José L. Murillo,a Rafael Cases,a Julian Carrey,d Carlos Estepa,a Cecilia Sosa,e
Fernando Palacio,a Luis D. Carlos,b
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a. ICMA, CSIC–Universidad de Zaragoza, 50009 Zaragoza (Spain).
b. Departamento de Física and CICECO, Universidade de Aveiro, 3810–193 Aveiro (Portugal)
c. Departamento de Electrónica de Potencia. I3A. Universidad de Zaragoza 50018 Zaragoza (Spain)
d. Université de Toulouse, INSA, UPS, CNRS (UMR 5215), F-31077, Toulouse (France)
e. Departamento de Toxicología, Universidad de Zaragoza, 50013 Zaragoza (Spain)
Magnetic- and light-induced thermal heating of nanoparticles are powerful non-invasive
techniques for bio and nanotechnology applications, such as hyperthermia therapy of can-
cer and other diseases. To be effective this local heating requires an adequate monitoring
of the nanoheaters local temperature. However, efficient and sensitive single nanoparticles
integrating heaters and thermometers are a real synthetic challenge. Here we present a
successful realization of this challenge. The nanoheaters are magnetic nanoparticles coated
with amphiphilic polymers that had been already used in magnetic hyperthermia [1]. The
ratiometric thermometric system was also developed in our group a few years ago [2]. The
temperature readout is optical and the thermometric probes are dual-emissive Eu3+/Tb3+
lanthanide complexes. These complexes are encapsulated in the hydrophobic inner part
of the nanoparticle coating, around the iron oxide magnetic core. The resulting heater/
thermometer nanoplatform shows an outstanding performance in terms of: low thermometer
heat capacitance (0.021·K-1) and heater/thermometer resistance (1 K·W-1); high temperature
sensitivity (5.8%×K-1 at 296 K) and uncertainty (0.5 K), physiological working temperature
range (295-315 K), readout reproducibility (>99.5%), and fast time response (0.250 s).
Experiments of time-resolved thermometry under an AC magnetic field reveal the existence
of an unexpected temperature gradient between nanoheaters and surrounding media for
relatively long time intervals (t > 10 s) and relatively low heat powers (10-16 W/heater). The
fitting of the data to simple heat models evidences the failure of using macroscopic thermal
parameters to describe heat diffusion at the nanoscale. A proof of concept of tempera-
ture mapping has been realized on cells that were incubated with the nanoparticles. The
fluorescence microscopy in two different wavelengths simultaneously after beam splitting
permits the mapping of the intracellular local temperature using the pixel-by-pixel ratio of
the Eu3+/Tb3+ intensities.
References1. Brites, C. D. S. et al. Adv. Mater. 22(2010), 4499.2. R. Bustamante et al. Phys. Rev. B 88(2013), 184406.
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19 .Magnetic hyperthermia, a promising tool for the minimal-invasive treatment of tumorsIngrid HilgerInstitute for Diagnostic and Interventional Radiology, Jena University Hospital, Erlanger Allee 101.
D-07747 Jena, Germany
Early detection of tumours has been distinctly improved in the last years, which favours the
development of minimal-invasive therapeutic methods promising a high therapeutic effi-
cacy. Herein, the treatment of small tumours via magnetic heating was proposed, meaning
the selective accumulation of magnetic materials, magnetite, at the tumour area and the
exposure of the breast to an alternating magnetic field for several minutes. This will pro-
duce a selective heating spot which allows for a localized elimination of tumour cells. Even
though the principle seems to be simple at a first glance, the efficacy of the methodology
will strongly depend on how the different biomedical, physical and technical parameters are
being combined and adjusted between each other. Therefore, in the present study we will
consider the different aspects that have to be taken into account to fulfil these requirements.
20 .Encapsulation of VEGF165 in magnetic PLGA nanocapsules for potential local delivery and bioactivity into human brain endothelial cellsRoig,1 E. Carenza1, O. Jordan2, P. Martínez-San Segundo3, R. Ji�ík4, Z. Star�uk jr.4, G. Borchard2,
A. Rosell3
1. Nanoparticles and Nanocomposites Group (www.icmab.es/nn), Institut de Ciència de Materials
de Barcelona, Consejo Superior de Investigaciones Científicas (ICMAB-CSIC), Campus de la UAB,
Bellaterra, Catalunya, E-08193 Spain.
2. School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Quai Ernest
Ansermet 30, 1205 Geneva, Switzerland.
3. Neurovascular Research Laboratory and Neurovascular Unit, Vall d’Hebron Institut de Recerca,
Universitat Autònoma de Barcelona; Passeig Vall d’Hebron, Barcelona, Catalunya, E-08035 Spain.
4. Institute of Scientific Instruments, Academy of Sciences of the Czech Republic, Královopolská 147,
612 64 Brno, Czech Republic.
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The administration of pro-angiogenic proteins is an attractive therapeutic strategy to enhance
angiogenesis after an ischemic event [1,2]. Nevertheless a labile structure and short circula-
tion time in vivo are the main obstacles that reduce protein bioactivity and dose at the target
site. Biodegradable magnetic nanomaterials, intravenously administrated and accumulated
at the diseased zone under an external magnetic field, may circumvent these limitations.
We report on the synthesis of poly(D,L-lactic-co-glycolic acid) (PLGA) nanocapsules (diam-
eter less than 200 nm) loaded with the vascular endothelial growth factor-165 (VEGF165)
confined into the inner core and superparamagnetic iron oxide nanoparticles (SPIONs)
embedded in the polymeric shell (Fig. 1 inset). The system showed high encapsulation
efficiencies and sustained protein release over 14 days (Fig. 1). In vitro studies confirmed
protein bioactivity after encapsulation. Moreover, through MRI measurements we demon-
strated excellent T2 contrast image properties. We therefore suggest the nanoconstruct as
new targeted protein delivery carrier useful in pro-angiogenic treatments, especially after
an ischemic event [3].
References1. Rosell, A.; Morancho, A.; Navarro-Sobrino, M.; Martinez-Saez, E.; Hernandez-Guillamon, M.; Lope-Piedrafita, S.;
Barcelo, V.; Borras, F.; Penalba, A.; Garcia-Bonilla, L.; Montaner, J., Factors secreted by endothelial progenitor cells enhance neurorepair responses after cerebral ischemia in mice. PloS one 2013, 8 (9), e73244.
2. Carenza, E.; Barceló, V; Morancho, A.; Lisa Levander, L.; Boada, C.; Laromaine, A.; Roig A.; Montaner, J.; Rosell, A., In vitro Angiogenic Performance and in vivo Brain Targeting of Magnetized Endothelial Progenitor Cells for Neurorepair Therapies. Nanomedicine: NBM 2014, 10, 1 225-234.
3. Carenza, E.; Jordan O.; San Segundo Martínez, P.; Ji�ík, R.; Star�uk jr., Z.; Borchard, G.; Anna Rosell, A.;* and Roig, A.*. Encapsulation of VEGF165 in magnetic PLGA nanocapsules for potential local delivery and bioactivity into human brain endothelial cells. J. Mater. Chem. B, 2015, Accepted Manuscript DOI: 10.1039/C4TB01895H
Figure 1: Protein release over a period of 2 weeks. Inset: scheme and cryo-TEM image of a PLGA
nanocapsule dispersed in water
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21 .Influence of nanoparticle size and field frequency on the concentration dependence of magnetic heating David Cabrera,a Gorka Salas,a Robert Ludwig,d Julio Camarero,a,b, Rodolfo Miranda a,b, Ingrid
Hilger,d and Francisco J. Terána, c
a. IMDEA Nanociencia, Ciudad Universitaria de Cantoblanco, 28049 Madrid, Spain ;
b. Dpto. Física de la Materia Condensada, Universidad Autónoma de Madrid, 28049 Madrid, Spain
c. Unidad Asociada de Nanobiotecnología, CNB-CSIC & IMDEA Nanociencia, 28049 Madrid, Spain;
d. Institute for Diagnostic and Interventional Radiology I, Jena University Hospital, Friederich Schiller
University of Jena, Bachstraße 18, D-07740 Jena, Germany;
Iron oxide nanoparticles (IONP) have attracted much attention for their potential in bio-
medical applications in the last years. Their precise size control and tailored chemical
features favor their internalization into cancer cells acting as drug-delivery nanocarriers
without cytotoxicity drawbacks. In addition, their size-driven magnetic properties allow
IONP to efficiently act as contrast agents, or intracellular hyperthermia mediators.
Recent works have shown that the internalization of IONP into cells and/or tissues lead
to significant changes of their magnetic properties, resulting in a remarkable reduction of
their heating efficiency [1]. The underlying reasons are yet to be clarified. At a first glance,
IONP processing into biological matrix (cells or tissues) increases IONP clustering favoring
magnetic dipolar interactions, which may be altered by viscosity. Hence, understanding
the role of dipolar interactions is of great importance towards the use of IONP as reliable
heating mediators.
Here, we report on the study of specific absorption rate (SAR) as a function of IONP con-
centration at different field frequencies and particle sizes. Our results indicate that both
parameters (size and frequency) lead to distinct concentration dependences matching within
the single picture described by Martinez-Boubeta et al [1]. While 19 nm size IONP show
a non-monotonic behaviour with a relative SAR maximum the 12 nm size IONP shows a
progressive decrease of SAR values until reaching a value which maintains constant in the
overall concentration range studied. When varying filed frequency, the variety of behaviours
becomes richer, resulting in the observation of an evolution from non-monotonic behaviour
of SAR vs concentration to the progressive SAR decay (at low frequencies). However, we
observe a shift of the relative SAR maximum which is clearly frequency-dependent and has
not been considered by recent theoretical models [2]. Our data provide new experimental
evidences which require novel theoretical models for understanding the role of magnetic
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dipolar interactions in determining the heat dissipation processes of IONP under alternat-
ing magnetic fields.
(a)
0 5 10 15 200
10203040
200
250
300
350
400
19 nm 12 nm
SAR
(W/g
Fe)
[Fe](gFe/L)
(b)
0 5 10 15 200
100200300400
1000
1500
2000
2500
3000
435 kHz 150 kHz 84 kHz 49 kHz
SAR(
W/g
Fe)
[Fe](gFe/L)
Figure 1. (a) Concentration dependence of SAR values obtained IONP water dispersions of different sizes (19 and 12 nm) at given field conditions (104 kHz and 50mT), (b) Concentra-tion dependence of SAR values obtained in 19±2 nm IONP water dispersions at different field frequencies and given field amplitude (40 mT)
References1. C. Martínez-Boubeta et al. Advanced Functional Materials 22 (2012) 37372. R. P. Tan, et al. Phys. Rev. B. , 35 (2014) 214421.
22 .Intratumoral thermal reading during photothermal therapy by multifunctional fluorescent nanoparticlesBlanca del Rosal,a Elisa Carrasco,b Francisco Sanz-Rodríguez,a,c,d Ángeles Juarranz de la
Fuente,b,c Patricia Haro-Gonzalez,1 Ueslen Rocha,e Kagola Upendra Kumar,e Carlos Jacinto,e
José García Solé,a and Daniel Jaque.a
a. Fluorescence Imaging Group. Departamento de Física de Materiales, Facultad de Ciencias. Campus
de Cantoblanco, Universidad Autónoma de Madrid. Madrid 28049, Spain
b. Instituto de Investigaciones Biomédicas “Alberto Sols”, CSIC-UAM. Madrid 28029, Spain
c. Departamento de Biología, Facultad de Ciencias. Campus de Cantoblanco. Universidad Autónoma
de Madrid. Madrid 28049, Spain
d. Instituto Ramón y Cajal de Investigación Sanitaria, Hospital Ramón y Cajal, 28034 Madrid, Spain
e. Grupo de Fotônica e Fluidos Complexos. Instituto de Física, Universidade Federal de Alagoas
57072-970, Maceió, Alagoas, Brazil
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Photothermal therapy, which relies on light-induced heating to irreversibly damage cancer
cells is nowadays attracting a great deal of attention as an effective and low cost technique
for treating malignant tumors.[1] An adequate temperature monitoring during the treatment
is key to minimizing damage to the surrounding healthy tissues while ensuring the effective-
ness of the therapy. However, measuring intratumoral temperature constitutes a challenge
that cannot be solved by traditional thermometry techniques.
Multifunctional nanoparticles, acting simultaneously as temperature sensors and photo-
thermal agents upon excitation with a single light source are excellent candidates for this
task. In this work, neodymium-doped LaF3 nanocrystals have been successfully used for
continuous intratumoral temperature monitoring during photothermal therapy in mice. These
infrared-emitting nanoparticles double as efficient in vivo heating agents and temperature
sensors. The use of this kind of nanoparticles for temperature-controlled therapy opens the
way for highly efficient and minimally invasive photothermal treatments of cancer tumors.
References1. D. Jaque et al. Nanoscale, 6 (2014) 9494.
23 .Magnetic nanoparticle-based therapeutic agents for thermo-chemotherapy treatment of cancerProf Nguyen TK ThanhUCL Healthcare Biomagnetic and Nanomaterials Laboratories
21 Albemarle Street, London W1S 4BS &
Department of Physics & Astronomy
University College London
Gower Street, London WC1E 6BT, UK
Email: [email protected]
http://www.ntk-thanh.co.uk
Magnetic nanoparticles have been widely investigated for their great potential as mediators
of heat for localised hyperthermia therapy. Nanocarriers have also attracted increasing
attention due to the possibility of delivering drugs at specific locations, therefore limit-
ing systematic effects. The enhancement of the anti-cancer effect of chemotherapy with
application of concurrent hyperthermia was noticed more than thirty years ago. However,
combining magnetic nanoparticles with molecules of drugs in the same nanoformulation
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has only recently emerged as a promising tool for the application of hyperthermia with com-
bined chemotherapy in the treatment of cancer. We will present some of our recent work
in the development of nanoparticles and effort in developing multifunctional therapeutic
nanosystems incorporating both magnetic nanoparticles and drugs.
.
References1. Blanco-Andujar, C., Southern, P., Ortega, D., Nesbitt, S.A., Pankhurst, Q.A., Thanh, N.T.K.* (2015) High per-
formance multi-core iron oxide nanoparticles for magnetic hyperthermia: microwave synthesis, and the role of core-to-core interactions. Nanoscale.. 7: 1768-1775. Open Access
2. A. Hervault and Thanh, N. T. K* (2014) Magnetic Nanoparticles-Based Therapeutic Agents for Thermo-Chemo-therapy Treatment of Cancer. Nanoscale, 6: 11553-11573. Open Access.
3. Thanh, N. T. K*, N. Maclean, S. Mahiddine. (2014) Mechanisms of Nucleation and Growth of Nanoparticles in Solution. Chemical Reviews. 114: 7610–7630. Open Access.
4. A. Dunn, D. Dunn, M. Lim, C. Boyer, N. T. K. Thanh. (2014) Recent Developments in the Design of Nanocom-posites for Photothermal and Hyperthermia Induced Controllable Drug Delivery. Royal Society of Chemistry Specialist Periodical Report on Nanoscience. 2: 225-254.
5. C. Blanco-Andujar, D. Ortega, Q. A. Pankhurst, N.T. K. Thanh* (2012) Elucidating the morphological and struc-tural evolution of iron oxide nanoparticles formed by sodium carbonate in aqueous medium. Journal of Material Chemistry. 22: 12498-12506 Open Access
24 .Nano-emulsion templating: a versatile technology to prepare multifunctional polymeric nanoparticles for advanced biomedical applicationsCristina Fornaguera, Gabriela Calderó, Conxita Solans Institute of Advanced Chemistry of Catalonia (IQAC-CSIC) and CIBER of Bioengineering, Biomaterials
and Nanomedicine (CIBER-BBB)
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Nano-emulsions are fine emulsions with droplet sizes between 20 – 200nm, showing high
kinetic stability and transparent to translucent aspect. Since they are thermodynamically
unstable systems, an energy input is required for their formation. Among the methods for
nano-emulsion preparation, low-energy methods, which take advantage of the chemical
energy of nano-emulsion components, are advantageous for pharmaceutical applications,
since they enable obtaining nano-emulsions with smaller and less polydisperse droplets.
Specifically, the phase inversion composition method (PIC), in which phase inversion is
produced by changing the composition during the emulsification process at a constant
temperature, is adequate for thermolabile compounds because it can be performed at mild
temperature and process conditions [1]. Polymeric nanoparticles (pNP) can be obtained
from nano-emulsions with an oil phase consisting on a preformed polymer dissolved in an
organic volatile solvent followed by solvent evaporation [2]. In this context, the objective of
this work was to obtain pNP, from nano-emulsion templating, as advanced pharmaceuti-
cal agents for diagnosis and/or therapeutic specific uses. O/W polymeric nano-emulsions
were firstly obtained by the PIC method, using poly-(lactic-co-glycolic acid) (PLGA) as the
preformed polymer [3,4]. They showed hydrodynamic radii between 20 – 140 nm, negative
surface charge and enough stability to prepare nanoparticles. Nano-emulsions with 90wt%
W, 70/30 O/S ratio were selected for further uses because they represent a compromise
between small sizes (around 40nm) and low surfactant concentration. pNP were prepared
from these nano-emulsions by solvent evaporation, resulting in hydrodynamic radii of around
20 nm, negative surface charge and stability higher than 3 months. These pNP were loaded
and /or functionalized to achieve different purposes. Fluorescent dyes (e.g. coumarin 6) or
inorganic nanoparticles (e.g. magnetic nanoparticles) were encapsulated in the pNP to use
them as diagnosis carriers, obtaining very high encapsulation efficiency. pNPs were also
designed as therapeutic agents following a pharmacological approach, in parallel to a gene
therapy approach. Model analgesic and antiapoptotic drugs were successfully encapsu-
lated to develop the pharmacological therapy, achieving also high encapsulation efficienciy
and sustained drug release, maintaining the pharmacological activity of the drugs. To use
nanoparticles as non-viral gene delivery systems, they were cationized to further electrostati-
cally bind antisense oligonucleotides. Very high gene transfection efficiencies were obtained
in cell cultures, with the advantage of pNPs over commercial vectors of a lower toxicity and
over viral vectors of lower immune detection. In addition, with the purpose to actively target a
specific organ, pNP surface was functionalized with a model monoclonal antibody targeting
the Blood-Brain Barrier (BBB). In vivo studies of the BBB crossing evidenced that antibody
functionalized pNP were able to cross the BBB in a similar extent than positive controls.
Therefore, it can be concluded that the preparation of polymeric nano-emulsions by low-
energy methods, followed by solvent evaporation to obtain polymeric nanoparticles is a very
versatile technology that enables tailoring their properties rendering them useful for both,
therapeutic and diagnosis purposes, achieving also an active targeting to the desired organ.
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References1. C. Solans et al., Curr Opin Colloid Interface Sci, 17 (2012) 246.2. C. Calderó et al., J Colloid Interface Sci, 353(2011) 406.3. C. Fornaguera et al., Colloid Surf B, 125(2015) 58.4. C. Fornaguera et al., Int J Pharm, 478(2015) 113.
25 .G-Quadruplex aptamers with therapeutic applicationsAnna Aviñóa, Santigo Grijalvoa, Carme Fàbregaa, Maria Tintoréa, Stefania Mazzinib, Juan
Carlos Moralesc and Ramon Eritjaa
a. Networking Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Institute
for Advanced Chemistry of Catalonia (IQAC), CSIC, Jordi Girona 18-26 08034 Barcelona, Spain
b. Department of Food, Environmental and Nutritional Sciences (DEFENS), Section of Chemical and
Biomolecular Sciences, University of Milan, Via Celoria 2, 20133 Milan, Italy.
c. Instituto de Investigaciones Químicas, CSIC - Universidad de Sevilla, Americo Vespucio, 49, 41092
Sevilla, Spain.
Aptamers are short DNA- or RNA-based oligonucleotides selected from large combinatorial
pools of sequences for their capacity to efficiently recognize targets, ranging from small mol-
ecules to proteins or nucleic acid structures. Like antibodies, they exhibit high specificity and
affinity for target binding. Aptamers can be considered as antibodies, but, unlike antibodies
they can be chemically derivatized to extend their lifetimes and/or their bioavailability and
are non-immunogenic. As a result, they may display effective interference in biological proc-
esses, which renders them not only valuable diagnostic tools, but also promising therapeutic
agents. Among several architectures, the polymorphic G-quadruplex structure is adopted by
multiple aptamers. G-quadruplexes are very stable structures generated by the association
of steps of four guanines (G-quartets) held together by hydrogen bonds. G-quadruplexes
acting as aptamers can be powerful inhibitors of different proteins. Particularly, large research
efforts have been focused on the 15-base long thrombin binding aptamer (TBA) sequence,
which was derived as an inhibitor of �-thrombin, a key enzyme in the blood clotting cascade.
In addition, several anti-HIV aptamers also adopt DNA quadruplex structures. Among them,
“Hotoda’s aptamer” has significant affinity to the HIV protein gp120. These aptamers are
potentially prone to be modified in order to improve biological properties.
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Figure 1. Struture of the TBA, G-tetrad and Hotoda.
References1. A. Aviñó, S. Mazzini, R. Ferreira, R. Eritja, Bioorg. Med. Chem., 18 (2010) 7348.2. H. Saneyoshi, S. Mazzini, A. Aviñó, G. Portella, C. González, M. Orozco, V. Marquez, R. Eritja, Nucleic Acids
Res., 37, (2009) 5589.3. J. Gros, A. Aviñó, J. López de la Osa, C. González, L. Lacroix, A. Pérez, M. Orozco, R. Eritja, J.L. Mergny. Chem.
Commun., 25 (2008) 2926.4. I. Gómez-Pinto, E. Vengut-Climent, R. Lucas, A. Aviñó, R. Eritja, C. González, J.C. Morales, Chem. Eur. J., 19,
(2013) 1920.5. A. Aviñó, S. Mazzini, R. Ferreira, R. Eritja, R. Gargallo, V. E. Marquez, R Eritja. Bioorg. Med. Chem., 20, (2012)
4186.
26 .Targeted Drug Delivery Systems against Cancer Stem CellsDr. Petra Gener Vall d’Hebron Research Institute,
Barcelona, Spain
Despite the progress in understanding the mechanism of carcinogenesis, and the develop-
ment of more effective therapies against cancer, advanced cancer is still remaining to be
an incurable disease. The cure of advanced cancer is hampered because of metastatic
spread to distal organs and resistance to therapy. Both phenomena are sustained by the
presence of cancer stem cells (CSC) within the tumor. This minor cell population retains the
capacity of spreading the tumor, while being insensitive to conventional anticancer therapies,
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antimitotic agents or radiation. Specific delivery of chemotherapeutic compounds to CSC
by nanotechnology based drug delivery systems (DDS) has great potential to increase CSC
sensitivity. To be able to study efficacy of nanomedicines in population of CSC we devel-
oped an original CSC model, in which we stably tagged CSC population. This CSC model
we used to study the impact of targeted/non-targeted DDS on CSC presence and behavior
in continuous cell culture.
27 .Development of gene-expressing liposomes as drug delivery systemsAlicia Soler Canton,a Christophe Danelonb
a. Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology,
Lorentzweg 1, 2628 CJ, Delft, The Netherlands
b. Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology,
Lorentzweg 1, 2628 CJ, Delft, The Netherlands
Targeted drug delivery in human body can be mediated by diverse nanocarriers. Among the
different types, liposomes – artificial phospholipid vesicles – have received the most atten-
tion during the last three decades for numerous reasons. Current liposomal systems deliver
therapeutic compounds that have been directly packaged upon manufacturing. However,
recent advances on the development of synthetic gene circuits prompt us to explore the
biomedical potential of gene-expressing liposomes. In this work we aim to express small
interfering RNA (siRNA) molecules of therapeutic value from a DNA template inside <
200 nm sized liposomes. Two main reasons drag us to work on the development of gene-
expressing liposomes for the production of therapeutic siRNA. First, we hypothesize that
the siRNA concentrations reached upon synthesis inside liposomes could be higher than
those obtained through the encapsulation of molecules produced in bulk (current method).
Second, we believe that future genetic-circuit-containing liposomes could manage a control-
led expression and release of the therapeutic molecule. Early steps towards the realization
of this new generation of programmable drug delivery systems include 1) the validation of
the production of functional shRNA by in vitro transcription and 2) the development of a
protocol for optimized transcription inside small unilamellar vesicles, which we have man-
aged so far inside 800nm liposomes. Later steps on this project will involve the merging
of these advances to transfect shRNA-expressing liposomes into cells and track specific
gene silencing.
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28 .Self-assembling nanoparticles based on calixarene-cyclodextrin heterodimers for targeted delivery and controlled release of docetaxelLaura Gallego-Yerga,a Francesco Sansone,b Carmen Ortiz Mellet,a Alessandro Casnatib and
José M. García Fernández.c
a. Departamento Química Orgánica, Facultad de Química, Universidad de Sevilla, E-41012 Sevilla
Spain;
b. Dipartimento di Chimica, Università degli Studi di Parma, I-43124 Parma, Italy;
c. Instituto de Investigaciones Químicas (IIQ), CSIC – Universidad de Sevilla, E-41092 Sevilla, Spain
The development of drug delivery systems able to improve bioavailability of insoluble or
biodegradable drugs and avoid secondary effects through targeted transport is essencial
for the success in administration of therapeutic agents. In this work we have succeeded at
preparing molecularly defined calixarene-cyclodextrin heterodimers filled with the capacity
to self-assemble in water or buffer media to form functional nanoparticles. These nanos-
tructures have an inner core formed by hydrophobic calix[4]arene (CA4)[1] units and an
external hydrophilic shell exposing b-cyclodextrin (bCD) moieties[2,3]. The CA4 scaffold is very
well suited to promote tight packing of fatty chains installed at the narrower ring in its cone
conformation, providing a lipid matrix where hydrophobic drugs can be entrapped, whereas
the presence of hydrophilic bCD moieties at the nanosphere surface allows nanoparticle
solubilization in water as well as ligand incorporation by inclusion complex formation. The
potential of the new systems in nanomedicine is illustrated by their capacity to encapsu-
late and provide sustained release of the anticancer agent docetaxel (DTX) and undergo
supramolecular surface post-modification with adamantane-armed glycoligands targeting
the human macrophage mannose receptor (MMR). Since docetaxel can induce apoptosis
in tumor cells which overexpress this receptor, this targeting is expected to improve cancer
immunotherapies.
References
1. Sansone, F.; Casnati, A. Chem. Soc. Rev. 2013, 42, 4623-4639.
2. Martínez, A.; Ortiz Mellet, C.; García Fernández, J. M. Chem. Soc. Rev. 2013, 42, 4746-4773.
3. Ortiz Mellet, C.; García Fernández, J. M.; Benito, J. M. Chem. Soc. Rev. 2011, 40, 1586-1608.
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29 .Design of biocompatible magnetic nanoplatforms for potential platinum-based drug delivery applicationsGustavo B. da Silva,a,b Rocío Costo,b Marzia Marciello,b Maria D. Vargas,a Célia M. Ronconi,a
M. Puerto Moralesb and Carlos J. Sernab
a. Instituto de Química, Universidade Federal Fluminense, Campus do Valonguinho, Centro, 24020-
150 Niterói-RJ, Brazil
b. Department of Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid
(ICMM)/CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
Cisplatin (cis-[Pt(NH3)2Cl2]) and its derivatives have been widely used in cancer treatment,
however they have shown several side effects. To overcome these drawbacks, the use of
magnetic nanoparticles as carriers and drug delivery systems could be a good strategy [1].
Iron oxide nanoparticles e. g. maghemite (g-Fe2O 3) are biocompatible, superparamagnetic
and can reach different regions of the body upon an external magnetic field. Furthermore,
they are easily modified to which platinum(II) derivatives can be attached and protected
against early deactivation, with consequent enhancement in their cytotoxicity.
Aim of this work was the synthesis of superparamagnetic g-Fe2O3 nanoparticles and sub-
sequent surface modification with a biocompatible polymer for the conjugation of cisplatin
derivatives with potential anticancer properties. The g-Fe2O3 nanoparticles were synthesized
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by co-precipitation method followed by an acid treatment [2]. Then they were coated with cit-
ric acid (CA) and amine dextran (ADex), through amide bonds. Investigation of the conditions
(iron and dextran concentrations, reactive type and pH) to best attach the ADex polymer to
the surface of g-Fe2O3 resulted in g-Fe2O3@CA@ADex containing ~70% of anchored polymer.
The materials were characterized by TEM, DRX, IR, TG-DTA and ICP-OES measurements.
Magnetic data confirmed a superparamagnetic core. The colloidal properties (hydrodynamic
size - DHyd, potential z) were evaluated and the stability in the physiological environment were
enhanced after functionalizations. Finally cisplatin related compounds (cis-[Pt(NH3)2(NO3)2]
and cis-[Pt(DMSO)2Cl2]) were attached to the ADex polymer to yield cis-[Pt(X)2(ADex)2], X =
NH3 or Cl, (25-45%). These platinum-modified polymers were then anchored to the surface
of g-Fe2O3@CA nanoparticles under the conditions employed for the synthesis of g-Fe2O3@
CA@ADex. Studies of colloidal stability and drug release are in progress.
References1. B. W. Harper et al. Chem. Eur. J., 16 (2010) 7064.2. R. Costo et al. Langmuir, 28 (2012) 178.
6 nm
biocompatibili ty
OO
OOHO
OH
O
HOHO
ONH
OHN
ADex-CA�-Fe2O3
ADexenhance
cell uptake
% coating = 14-70%DHyd = 100-200 nm
Msat = 65 emu.g-1
of nanoparticle
% coating = 10%DHyd = 27 nm
Pt
PtPt
Pt
PtACMDex
ACMDex
NH3
NH3
PtACMDex
ACMDex
ClCl
or
cisplat in der ivatives
O
HO OH
O
OH
OHO
Citric acid (CA)
ADexmodified
with cisplatinderivatives
Magneticnanoplatforms
Lack of stabilityat pH 7.0
�-Fe2O3�-Fe2O3@CA
�-Fe2O3@CA@ADex�-Fe2O3@CA@ADex-Pt
Figure 1. General route for design of magnetic nanoparticles of iron oxide (g-Fe2O3) and its modi-
fication with citric acid, amine dextran and cisplatin derivatives.
POSTER contr ibut ion
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P1.
Interaction of multifunctional magnetic nanoparticles with biological membranes
María Josefa Rodrígueza, Marzia Marciellob, Antonio Airesc, Sandra M. Ocampoc, Aitziber L. Cortajarenac, Puerto M. Moralesb, José L. Carrascosaa
a Department of Structure of Macromolecules, Centro Nacional de Biotecnología-CSIC (CNB-CSIC), Campus Cantoblanco, 28049 Madrid, Spain
b Instituto de Ciencia de Materiales de Madrid (ICMM)-CSIC, Campus Cantoblanco, 28049 Madrid, Spain
c Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia). Cantoblanco, Madrid, Spain.
Abstract
The possibility to selectively introduce magnetic nanoparticles (MNPs) inside cells for further manipulation is a key objective in nanomedicine. To achieve this purpose, MNPs need to overcome a first barrier represented by biological membranes, which also define the selectivity characteristics of the target cells. The biological functionalization of the MNPs will enhance the selectivity toward the desired cells, improving the efficiency of incorporation of therapeutic molecules and reducing the effects. In this study, imaging of thin sections of cultured cells by transmission electron microscopy (TEM) have allowed us to evaluate at an ultrastructural level the interaction of MNPs with cellular membranes and the characteristics of uptaking and accumulation inside the cells of nanoparticles with different coatings and functionalizations. In particular, MNPs with two different coatings, a small molecule (dimercaptosuccinic acid, DMSA) and a biocompatible polymer (polyethylene glycol, PEG) have been studied. We have found that the different nature of the coating and the functionalization influence nanoparticle biodistribution in cellular membranes and entry characteristics.
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P2.P1 .
Biodistribution of magnetic nanoparticles evaluated by AC magnetic susceptibility in a murine model
Eduardo Lorente-Sorolla,a Gustavo B. da Silva,a,b Sonia Perez-Yagüe,c Rocío Costo,a Sabino Veintemillas-Verdaguer,a Domingo F. Barber,c M. Puerto Morales,a Marina Talelli c and Lucía Gutiérrez a
a Department of Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid (ICMM)/CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
b Instituto de Química, Universidade Federal Fluminense, Campus do Valonguinho, Centro, 24020-150, Niterói, RJ, Brasil
c Department of Immunology and Oncology, and NanoBiomedicine Initiative, Centro Nacional de Biotecnología (CNB)/CSIC, Darwin 3, Cantoblanco, 28049 Madrid, Spain
Abstract
Iron oxide nanoparticles for biomedical applications preferentially accumulate in the liver and spleen, therefore reducing their possible uses in other organs, unless external magnetic fields or other functional molecules are used to target the particles to specific sites.
It is known that in thalassemia patients undergoing chronic blood transfusions, the excess iron from the blood cannot be properly bound to transferrin, leading to accumulation in the heart though the formation of iron-citrate complexes called NTBI (non-transferrin bound iron), as in the heart the metal-ion transporter ZIP14 is present, which mediates NTBI uptake [1]. The aim of this project was to mimic this NTBI function using citrate coated magnetic nanoparticles, in order to generate bio-inspired materials that may target the nanoparticles to the heart, therefore changing their biodistribution pattern.
We have performed a single administration of magnetic nanoparticles coated with citric acid and phosphonoacetic acid in mice and evaluated the presence of these nanoparticles in the heart, liver, lungs, spleen and kidney 24h after their administration. Nanoparticles with both coatings were clearly observed by AC magnetic susceptibility measurements in the liver, lungs and spleen tissues. The presence of nanoparticles in the heart was very small in comparison to the other tissues. Transmission electron microscopy in liver sections showed the presence of magnetic nanoparticles inside lysosomal structures. In addition, ferritin, the iron storage protein, was frequently observed localized near the lysosomes containing the magnetic nanoparticles in the liver tissues.
References
[1] J.P. Liuzzi et al. Proc. Nat. Acad.Sci. 103 (2006) 13612.
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Figure 1. (Left) Transmission electron micrograph of a liver tissue section showing the presence of magnetic nanoparticles and ferritin. (Right) Temperature dependence of the out-of-phase susceptibility of different tissue samples from the same mouse showing a maximum corresponding to the presence of
magnetic nanoparticles.
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P3.
C. elegans: an in vivo model to evaluate nanoparticles?
Anna Roig, Laura Gonzalez-Moragas, Elisa Carenza, Siming Yu, Anna Laromaine
Nanoparticles and Nanocomposites Group, Institut de Ciència de Materials de Barcelona, Consejo Superior de Investigaciones Científicas (ICMAB-CSIC), Campus de la UAB, Bellaterra, Catalunya, E-08193 Spain.
Abstract
Caenorhabditis elegans (C. elegans) is a 1-mm long free-living soil nematode widely used in biology as a model organism. The key attributes that make C. elegans a promising in vivo animal model to evaluate nanoparticles include simplicity, transparency, short life cycle, sequenced genome, small body size and ease of cultivation in the lab. Here, we investigate the effect of surface functionalization (citrate and protein-coated) of superparamagnetic iron oxide nanoparticles in C. elegans combining techniques from materials science and biology confirming the worm as a suitable in vivo platform to advance in the understanding of nano-bio-interfaces (Figure 1). The survival and nanoparticle distribution of worms treated with the two nanoparticles was studied. By using magnetometry, quantitative uptake and magnetic properties of the nanoparticles inside the worms were evaluated. Interestingly, results indicate the protective effects both to the protein to the nanoparticles and to the worms especially at high concentrations. Thus, our results endorse the potential of C. elegans to assess nanomaterials at early stages of their synthetic formulations and to reduce the number of candidate materials to be screened in mammalian models.
Figure1. Visualization of a C. elegans: Prussian Blue stained worm where iron oxide nanoparticles appear blue.
50 µm
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P4.P3 .
Iron quantification inside cells incubated with SPION by Soft-Xray Absorption Spectro-Tomography (SXAST)
José Javier Conesa,a Francisco Javier Chichón,a Michele Chiappi,a Eva Pereiro,b and José L. Carrascosa.a,c
a Centro Nacional de Biotecnología (CNB-CSIC), Cantoblanco, 28049 Madrid, Spain. b ALBA Synchrotron Light Source. MISTRAL Beamline - Experiments division. 08290 Cerdanyola del
Vallès, Barcelona, Spain. c Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), Cantoblanco, 28049
Madrid, Spain
Abstract
One of the mayor issues related to Superparamagnetic nanoparticles (SPION) internalization is the estimation of the amount of nanoparticles that are inside the cells. The answer to this question will determine the dose for drug-delivery or whetherhyperthermia treatments are feasible. To get an insight into this problem we used 15 nm DMSA-SPIONs interacting with MCF-7 breast cancer cells [1] as an experimental model, and Soft-X-ray spectral-microscopy to calculate the iron content -main component of the SPION- using the specific absorption of the element.
X-ray absorption is a widely used technique that exploits the singular interaction of the elements with X-rays [2]. For this study we developed a new approach called Soft X-ray Absorption Spectro-Tomography (SXAST) acquiring spectral tilted series of the same cell between at 700 and 709 eV. In this context, the iron abortion changes dramatically from 700 to 709 eV, while the rest of the cellular compound remains with the same contrast (Figure). We used this property to subtract the images corresponding to the tilted series acquired at 700 from the ones form the 709 eV tilted series. The result is a differential tilted series where the cellular background was eliminated. Then, tomographic reconstructions allowed us to obtain iron absorption coefficients, hence the iron mass, associated to three-dimensional cellular ultrastructure (Figure).
References [1] M.Calero, M.Chiappi, et al., “Characterization of interaction of magnetic nanoparticles with breast cancer cells”. Journal: Journal of Nanobiotechnology “in press”. [2] .Henke, B. L., E. M. Gullikson and J. C. Davis (1993). "X-Ray Interactions: Photoabsorption, Scattering, Transmission, and Reflection at E = 50-30,000 eV, Z = 1-92." Atomic Data and Nuclear Data Tables 54(2): 181-342.
700 eV 709 eV Fe signal image Fe signal slice
Figure. Soft-X ray Absortion Spectro-Tomography (SXAST) workflow. First two images represent 0º projection images of a dehydrated MCF7 cell incubated with OD15-DMSA SPION using Fe pre edge and edge energies (700 and 709 eV respectively). Alignment of projections at each different energy yield ratio images that must be aligned and then submitted to tomographic reconstruction (fourth image, central slice of the quantitative volume). Modified EFTEM TOMOJ software was used to create differential maps and reconstruction.
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P5.
Following the transformation of magnetic nanoparticles over time in macrophages by AC magnetic susceptibility
Vanesa del Dedo,a Marina Talelli,b José M. Rojas,b Gustavo B. da Silva,a,c Rocío Costo,a Sabino Veintemillas-Verdaguer,a Domingo F. Barber,b M. Puerto Morales,a Marzia Marciello a and Lucía Gutiérrez a
a Department of Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid (ICMM)/CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
b Department of Immunology and Oncology, and NanoBiomedicine Initiative, Centro Nacional de Biotecnología (CNB)/CSIC, Darwin 3, Cantoblanco, 28049 Madrid, Spain
c Instituto de Química, Universidade Federal Fluminense, Campus do Valonguinho, Centro, 24020-150, Niterói, RJ, Brasil
Abstract
Little is known about the degradation processes that magnetic nanoparticles (MNPs) undergo after their administration, which can give insight on their potential side effects when used for biomedical applications. A promising approach to follow the MNPs transformations in tissues is the use of AC magnetic susceptibility measurements [1,2]. However, once the nanoparticles start to transform in the body, the analysis of the magnetic susceptibility data for quantification purposes is not straightforward, as a reduction in the particle size cannot be easily distinguished from a disaggregation process that can also reduce inter-particle interactions.
To gain insights into the transformations of magnetic nanoparticles in biological systems, and therefore be able to better understand the changes of their magnetic properties, we followed their degradation in a macrophage cell line (THP1) for 5 days. Transmission Electron Microscopy (TEM) and AC magnetic susceptibility measurements have been performed at different time points after treatment of the cells with the nanoparticles. Results indicated that a partial degradation of the nanoparticles occurs during this time period, with a shift of the particle size distribution towards smaller particles. Further reduction of the particle sizes may require longer incubation times, however, this will require optimization of culture conditions.
References
[1] R. Mejías et al. J. Controlled Release, 171 (2013) 225.
[2] L. Gutiérrez et al. Phys.Chem.Chem.Phys.,16 (2014) 4456.
Figure 1. (Left and center) Transmission electron micrographs at two different magnifications of a THP1 cell line incubated with magnetic nanoparticles. (Right) Temperature dependence of the out-of-phase
susceptibility of cells incubated with the MNPs at different times after the administration.
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P6.P5 .
pH-Dependent Switch for Anticancer Metallodrugs
Francisco Martínez, Ana M. Pizarro
IMDEA Nanociencia, Ciudad Universitaria de Cantoblanco, 28049, Madrid
Abstract The successful development of platinum-containing anticancer drugs that began with cisplatin (cis-diamminedichloridoplatinum(II)) has triggered the investigation of other metal-based agents with cytotoxic activity for drug design. Ruthenium, osmium, iridium and rhodium organometallic complexes have been in the last few years one of the most promising findings due to their inherent anticancer activity and to the possibility of a different cytotoxic profile than platinum-based drugs. In general, ruthenium(II) arene-complexes of type [(η6-arene)RuII(XY)Z]n+ (Z = monodentate ligand; XY = chelating ligand such as ethylenediamine, en) are cytotoxic to cancer cells including cisplatin-resistant cell lines. Recently, new ruthenium(II) arene compounds with a hemilabile amino-derivatised arene ligand have been published,1,2 where the amino group offers two reversible functionalities: (i) binding to the RuII center to form a tether-ring-closed (inactive) complex, or (ii) dissociation of the amine from the RuII center (as a dangling arm) to afford an open-tether complex with a pendant free amino group. In the open form the vacant site on the ruthenium is occupied by either chloride, a solvent molecule or a biomolecule (active complex). Most importantly, in aqueous solution, the tether-ring dynamics are pH dependent, giving the potential to finely tune the activation process to biological conditions, for example, the acidic microenvironment of the tumor.3 In the present work, we show a series of ruthenium(II) “tethered” arene complexes bearing different hemilabile amino-derivatised arene ligands. They have been characterized by NMR and elemental analysis. The understanding of the conditions and kinetics that control the opening of this tether would be crucial for rationally designing drugs with controlled biological target recognition, enhanced cytotoxic activity and minimized off-target effects. The aquation and anation phenomena for this type of compounds are under investigation to finely tailor the structures where the opening-closing pKa is most suitable for switching on in tumours, sparing healthy cells and therefore optimising selectivity and efficacy. Finally, the extraordinary versatility of the metallodrug structure is amenable for engrafting onto nanoparticle carriers. Nanoparticles as drug delivery systems have received much attention due to their elegant capabilities to target and accumulate in tumour tissues as well as drop deadly cargoes in a more efficient, controlled, on-target manner.4 For example, the bidentate chelating ligand in the metal complexes can be modified to carry groups that can react with suitable functionalities on the surface of the selected nanoparticles.
References
[1] M. Melchart et al. Inorg. Chem, 46 (2007) 8950
[2] A. M. Pizarro et al. Inorg. Chem, 49 (2010) 3310
[3] D. Neri et al. Nat. Rev. Drug. Discov., 10 (2011) 767
[4] X. Wang. Chem. Soc. Rev, 42 (2013) 202
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P7.
Superparamagnetic properties of hydrothermally prepared CoFe2O4 as a function of size (6–10 nm) and coating (oleic/citric acid or TiO2)
Daniel Nižnanský,a Anton Repko,a Jana Vejpravová,b and Taťána Vacková c
a Charles University, Faculty of Science, Department of Inorganic Chemistry, Hlavova 2030/8, 128 43 Prague 2, Czech Republic
b Institute of Physics AS CR, v.v.i., Department of Magnetic Nanosystems, Na Slovance 2, 182 21 Prague 8, Czech Republic
c Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, 162 06 Prague 6, Czech Republic
email: [email protected]
Abstract
Magnetic properties of cobalt ferrite nanoparticles prepared by hydrothermal hydrolysis of Co-Fe oleate in the presence of pentanol/octanol/toluene and water at 180 or 220 °C were prepared. The particle size (6.2, 9.1 or 10.5 nm) was controlled by the organic solvent and temperature. The inter-particle distance was then changed by a surface modification with citric acid or titanium dioxide.
The as-prepared hydrophobic nanoparticles (coated by oleic acid) had an inter-particle distance of 2.5 nm. Their blocking temperature (estimated as a maximum of the zero-field-cooled magnetization) was 179 K, 283 K and 331 K. Replacement of the oleic acid by citric acid imparted hydrophilicity and the inter-particle distance decreased to almost zero, while the blocking temperature increased by approximately 10 K, as a consequence of stronger dipolar inter-particle interactions.
Another modification was achieved by coating with titanium dioxide, supported by nitrilotris(methylphosphonic acid). The increased inter-particle distance implied lowering of the blocking temperature by ca. 20 K. The CoFe2O4@TiO2 nanoparticles were sufficiently stable in water, methanol and ethanol.
Magnetic structure was also investigated by Mössbauer spectroscopy. The effect of dipolar interactions on relaxation phenomena of the nanoparticles of different size and coating was finally studied by alternating-current (AC) susceptibility measurements.
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P8.P7 .
Targeting cancer cells with photoactive silica nanoparticles
Wioleta Borzęcka a, b, c, Tito Trindade b, Tomás Torres c and João Tomé a, d
a QOPNA and b CICECO, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal c Department of Organic Chemistry, Autonoma University of Madrid, 28049 Madrid, Spain
d Department of Organic and Macromolecular Chemistry, Ghent University, B-9000 Gent, Belgium
Abstract
Photodynamic therapy (PDT) is an emergent cancer medical treatment that combines a nontoxic drug, a particular type of light and oxygen. This drug, called photosensitizer (PS) or photosensitizing agent, when in contact with molecular oxygen and exposed to light can produce reactive oxygen species (ROS) that are strongly cytotoxic to cancer cells. An ideal PS should be a single pure compound with high absorption peak between 600 and 800 nm (red to deep red), should have no dark toxicity, and relatively rapid clearance from normal tissues [1].
Like other clinical protocols, PDT also has some limitations. To avoid some of these disadvantages, different PS nanoformulations have been tried. This target strategy can improve PDT treatments by increasing the aqueous solubility of the hydrophobic photosensitizers, their blood circulation, and their selective accumulation in tumor tissue, thanks to the enhanced permeability and retention effect [2]. In this field, silica nanoparticles (SNPs) have recently emerged as promising vehicles for PDT owing to their: high biocompatibility, controllable size formation, unique physicochemical properties, the possibility for tumor targeting through facile surface modification [3].
Some of our recent work on smart porphyrin-SNP materials, presenting their synthetic strategies and obtained photo-chemical and -physical results will be highlighted.
References
[1] P. Agostinis; K. Berg; K. A. Cengel; T. H. Foster; A. W. Girotti; S. O. Gollnick; S. M. Hahn; M. R. Hamblin; A. Juzeniene; D. Kessel; M. Korbelik; J. Moan; P. Mroz; D. Nowis; J. Piette; B. C. Wilson; J. Golab A Cancer J Clin, 61 (2011) 250
[2] P. Couleaud; V. Morosini; C. Frochot; S. Richeter; L. Raehma J. O. Durand Nanoscale, 2 (2010) 1083
[3] L. Tang; J. Cheng Nano Today, 8 (2013) 290
[4] F. Figueira; J. A. S. Cavaleiro; J. P.C. Tomé J. Porphyrins Phthalocyanines, 15 (2011), 517
Figure 1. Differente methodes for the synthesis of nanoparticles [4].
Acknowledgements: Thanks are due to the Universities of Aveiro and Autonoma of Madrid, FCT, European Union, QREN, FEDER and COMPETE for funding the QOPNA research unit (PEst-C/QUI/UI0062/2013; FCOMP-01-0124-FEDER-037296), CICECO Laboratory (Pest-C/CTM/LA0011/2013, FCOMP-01-0124-FEDER-037271). Thanks are also due to the Seventh Framework Programme (FP7-People-2012-ITN) for funding the SO2S project (grant agreement number: 316975).
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P9.
Combinational sensitization of tumor cells with two photosensitizers synergistically enhances their photodynamic inactivation in vitro
Andrea Tabero,a Ana Lazaro-Carrillo,a,b Pilar Acedo,c Macarena Calero,a,b Juan Carlos Stockert,a Magdalena Cañete,a Angeles Villanueva a,b
a Departamento de Biología. Universidad Autónoma de Madrid. Spain b Madrid Institute for Advanced Studies in Nanoscience (IMDEA). Spain
c Karolinska Institute. Stockholm. Sweden.
Abstract Liposomal lipid-based nanocarriers are widely used in drug delivery, especially in the treatment of cancer disease. Photodynamic therapy (PDT) is a minimally invasive and clinically approved procedure for cancer treatment. It can be defined as the administration of a drug known as photosensitizer (PS), which preferentially accumulates in tumor cells and that does not induce a cytotoxic effect by itself. However, after being irradiated with light of appropriate wavelength, the PS is able to trigger photochemical and photobiological reactions that lead to the generation of reactive oxygen species (ROS, mainly singlet oxygen), which cause irreparable damage, cell death and tumor regression. Our work is focused on a new photodynamic treatment based on the combination of two PSs: the hydrophobic Zinc(II)-phthalocyanine (ZnPc) incorporated into nanoliposomes and the hydrophilic cationic porphyrin meso-tetrakis(4-N-methylpyridyl)porphine (TMPyP). These PS have different subcellular location (Golgi apparatus for ZnPc and lysosomes for TMPyP) and therefore ROS are generated simultaneously in two different subcellular targets, which induces a synergistic effect using very low doses of PDT in human adenocarcinoma HeLa cell line. Cytotoxic MTT assay showed that the combined treatment has a powerful cell inactivation effect (cell death > 95%), while photodynamic treatments with each PS alone did not significantly affect cell survival. Using different morphological and biochemical analysis we have demonstrated that combined treatment triggers apoptotic cell death via mitochondrial-related pathway (1). On the other hand, caspase-3 is an important effector protease that cleaves many proteins in the apoptotic pathway. For this reason, we also analyzed the response to this treatment in human breast carcinoma cells (MCF-7), which do not express caspase-3. Our results revealed that this new strategy of photodynamic treatment induces over 95% MCF-7 cell death 24 h after treatment by a caspase 3-independent apoptotic pathway. In summary, combined PDT with ZnPc and TMPyP increases the ability to inactivate human tumor cells through apoptosis, even if they have activated a mechanism of evasion of this type of cell death, such as the caspase-3 deficiency. Our results provide a novel and valuable information that will contribute to the development of better PDT approaches for the treatment of various cancers.
References
[1] Acedo P, Stockert JC, Cañete M, Villanueva A. Two combined photosensitizers: a goal for more effective photodynamic therapy of cancer. Cell Death Dis. 2014, e1122. doi: 10.1038/cddis.2014.77.
This work was supported by the Spanish Ministry of Economy and Competitiveness (CTQ2013-48767-C3-3-R).
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P10.P9 .
Synthesis Strategies of Single-Core Magnetic Nanoparticles
Helena Gavilán,a Rocío Costo,a Sabino Veitemillas- Verdaguer,a Lucía Gutiérrez,a and M. Puerto Moralesa
Dept. Biomaterials and Bioinspired Materials, Instituto de Ciencia de Materiales de Madrid ICMM/CSIC, Sor Juana Inés de la Cruz 3, Cantoblanco, 28049 Madrid, Spain
Among other functional nanomaterials, magnetic iron oxide nanoparticles (MNPs) are especially promising as contrast enhancement agents for magnetic resonance imaging (MRI), drug carriers and for cancer treatment by hyperthermia. For all these biomedical applications, MNPs require among others, good magnetic properties, biocompatibility and stability. The former is deeply influenced by the crystal structure of the iron oxide nanoparticles and the magnetic interaction between the particles. It is essential therefore to control such characteristics.
Different synthesis strategies have been developed considering the fact that magnetic nanoparticles tend to form aggregates in the absence of specific coatings. There are mainly two kinds of systems that have been defined: multi-core and single-core particles.1 The former is composed of several magnetic cores per particle and although it is generally easy to synthesize such particles, it is difficult to control the number of cores per particle. The latter contains just one core per particle and nowadays there are just few coating methods that can be used to prevent aggregation by minimizing the inter-particle interactions.
Various synthesis strategies can be used to obtain these systems, being thermal decomposition in aqueous or organic solutions the most common one.2 Although thermal decomposition in organic media prevent the agglomeration of the particles as they are formed, it requires also complicated operations and sometimes expensive/toxic reagents. Nevertheless these particles possess high crystallinity and monodispersity and are the ideal material for standardization purposes.
As part of the NanoMag-project, we have prepared both single-core and multi-core magnetic particles. The iron oxide cores have been synthesized by thermal decomposition of iron oleate complex, and subsequently coated either individually with silica via microemulsion, or forming clusters with 2,3-dimercaptosuccinic acid (DMSA) by ligand exchange, being both colloids stable in water (essential pre-requisite for a biomedical application). In addition, we present a three-step aqueous approach to obtain Fe3O4 single-core particles based on the synthesis of antiferromagnetic nanoparticles, its coating and subsequent reduction to magnetite. This method has the advantage of the low inter-particle interactions of as-synthesized nanoparticles and the different morphologies that these materials exhibit.
References
[1] L. Gutiérrez et al., Dalton Trans, In press (2015)
[2] Xin Liang et al., Adv. Funct. Mater., 16, 1805 (2006)
Figure 1. Iron oxide nanoparticles: (A) Fe3O4 NPs synthesized by thermal decomposition in organic media. (B) Fe3O4 NPs coated with silica by microemulsion process. (C) α-FeOOH NPs synthesized by thermal decomposition in aqueous media. (D) β-FeOOH NPs synthesized by thermal decomposition in aqueous media.
A B C D
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P11.
Silica encapsulation of magnetic nanoparticles
Leonor de la Cueva,a Paloma Rodríguez,a Rebeca Amaro,a David Cabrera,a María Acebrón,a Beatriz H. Juárez,a,b Francisco J. Terán,a and Gorka Salasa
a IMDEA Nanociencia b Departamento de Química-Física Aplicada, Universidad Autónoma de Madrid
e-mail [email protected]
Abstract
Iron oxide nanoparticles (IONPs) based on magnetite and maghemite are promising tools for biomedical applications in treatment and diagnosis of tumours.[1] For treatment they can be employed as selective drug carriers or as nano-heaters. The latter application is known as magnetic hyperthermia and is based on the ability of these materials to dissipate heat under the influence of an alternating (AC) magnetic field.[2] However, there exist some problems limiting the use of IONPs as hyperthermia mediators in in vivo tissues. It has been observed that once into the cells the heat generation ability is seriously harmed mainly due to aggregation caused by the intracellular environment and processing.[3] This aggregation leads to magnetic dipolar interactions between particles that may worsen their heat dissipation power. One plausible approach to avoid this problem is to encapsulate the individual IONPs in a stable and rigid material to prevent those destructive interactions. Alternatively, a controlled aggregation of IONPs in an ordered fashion may enhance their magneto-thermal capacity, instead of damaging it.
In this work we show the encapsulation of individual IONPs in silica (IONP@SiO2 core@shell nanostructures) with controlled thickness, along with magnetic measurements comparing IONP@SiO2 with the initial non-encapsulated nanoparticles. Preliminary results on the encapsulation of nano-assemblies of IONPs will be also presented.
References
[1] M. Colombo et al. Chem. Soc. Rev. 41 (2012) 4306.
[2] S. Dutz and R. Hergt Int. J. Hypertherm. 29 (2013) 790.
[3] J. P. Fortin et al. Eur. Biophys. J. 37 (2008) 223; Q. A. Riegler, J. et al. Biomaterials 34 (2013) 1987.
Figure 1. Individual IONPs encapsulated with a silica shell.
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P12.P11 .
Synthesis of hybrid magneto-‐plasmonic nanostructures based on Au nanorods and iron oxides nanoparticles
Jesús G. Ovejeroa,b, Eva Mazariob, Alexandra Muñozb, Patricia Crespoa and Pilar Herrastib
aInstituto de Magnetismo Aplicado Salvador Velayos (UCM) P.O. Box 155, 28230 Las Rozas, Madrid, Spain bDepartamento de Química Física Aplicada, Universidad Autónoma de Madrid, 28049, Cantoblanco, Madrid, Spain
Abstract
Magnetic and plasmonic nanoparticles have brought a large number of biomedical solutions in the fields of detection, diagnosis and treatment. Both kinds of nanoparticles present their own limitations, but a question arises: Can we get unexpected synergetic properties in hybrid systems?
Surface plasmon resonance of Au nanoparticles is commonly used in biological sensing (SERS), medical imaging techniques (PAI, OCT, X-‐Ray, etc.) and in certain therapies (Photo-‐hyperthermia, thermally enhanced drug delivery, etc.). Moreover, different geometries of Au nanoparticles have been developed with the aim of shifting the plasmon resonance to the near infrared biological absorption window. Silica coated nanorods (Au@SiO NR) stand out from the rest of geometries due to their chemical stability and optimum optical properties [1],[2].
O the other hand, magnetic nanoparticles (MNP) have been used as alternative approach in fields such (MRI, MPI, magnetic hyperthermia, etc.). Thus, the combination of magnetic and plasmonic properties in a single nanoparticle offers, at first approximation, an interesting platform for multimodal imaging or a combined magneto-‐optic therapies. In addition, magnetomotive effect has been successfully used to improve the resolution of plasmon-‐based imaging techniques [3,4].
This work presents novel hybrid magneto-‐plasmonic nanoparticles based on the coupling of Au@SiO NR with different types of ferrite-‐based magnetic nanoparticles (MNP). Both, the Au@SiO NR and the MNP are synthesized separately and coupled afterwards. The followed process allows the tailoring of the magnetic and the plasmonic properties independently. Finally, the magneto-‐plasmonic nanoprobes have been functionalized with folic acid in order to improve their biocompatibility and internalization in cancer cells which overexpress folate receptors.
References
[1] Chen-‐Wei Wei et al. J.Biomed. Opt. 2010, 15(1):016010 [2] Laura M. Maestro et al. Langmuir 2014, 30: 1650 [3] R. Jhon et al. Proc. Natl. Acad. Sci U S A. 2010 107(18): 8085 [4] Y. Jin et al. Nature Comm. 1:41
This work has been supported by MAT2012-‐37109-‐C02-‐01 project of the Spanish Ministery of Economy and Competitivity
Figure 1. TEM image of Au@SiO NR coupled with Fe3O4
MNP
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P13.
Functionalized magnetic nanoparticles for cancer therapy
Antonio Aires,a,b Sandra M. Ocampo,a David Cabrera,a Leonor de la Cueva,a Yael Fernandez,a Gorka Salas a,b, Francisco J. Terán a,b, and Aitziber L. Cortajarena.a,b
aInstituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia).
b CNB-CSIC-IMDEA Nanociencia Associated Unit, Cantoblanco, Madrid, Spain.
email address of the Presenting Author: [email protected]
Abstract
In recent years, magnetic nanoparticles (MNP) have been widely investigated for their potential in biomedical applications acting as contrast agents for magnetic resonance imaging (MRI),1 nanocarriers for selective targeting and drug/gene-delivery,2 and as magnetic heating inductors for thermal therapeutic approaches.3 For successful in vivo application, MNP have to satisfy several requirements such as biocompatibility, invisibility to the immune system to avoid fast liver clearance, high colloidal stability and long blood circulation time. To achieve a long-term colloidal stability and long blood circulation time of MNP, different polymers coatings including poly(ethylene glycol) (PEG), dextran, chitosan, poly(ethylenimine) (PEI), and dimercaptosuccinic acid (DMSA), have been employed over the last years. Among the commonly-used polymer materials, proteins or polypeptides are considered to be among the most promising materials as protective layers of MNP due to their biocompatibility and hydrophilicity.4
In this study, we have developed MNP functionalized with an anti-cancer drug, in particular gemcitabine (GEM) to target pancreatic cancer. In order to achieve formulations with improved properties for in vivo use in drug delivery and hyperthermia treatment we developed a formulation in which the MNP are previously coated with BSA to provide enhanced colloidal stability in biological environments and long blood circulation time to the final formulation. The effect of surface functionalization on the colloidal stability in different media, magnetic properties, internalization behavior and in vitro cytotoxic effect in pancreatic cancer cells were investigated for their potential in vivo application in controlled drug release and/or magnetic hyperthermia.
References
[1] A. Masoudi et al. Int. J. Pharm. 433 (2012) 129. [2] N. S. Barakat et al. Nanomedicine, 4 (2009) 799. [3] E.K. Wang et al. Biomaterials, 30 (2009) 1881. [4] Z. Li et al. Colloids and Surfaces A: Physicochem. Eng. Aspects, 436 (2013) 1145.
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P14.P13 .
Detection and Inhibition of Mutated GNAQ Gene Using Spherical Nucleic Acids
Ana Latorrea, Christian Poschb, Alfonso Latorrea, Michelle B. Crosbyb, Anna Cellib, Igor Vujicb, Martina Sanlorenzob, Gary A. Greenb, Jingly Weierb, Mitchell Zekhtserb, Jeffrey Mab, Gabriela Monicob, Devron
H. Charb, Denis Jusufbegovicb, Klemens Rappersbergerb, Susana Ortiz-Urdab. Álvaro Somozaa
a Instituto Madrileño de Estudios Avanzados en Nanociencia (IMDEA Nanociencia), & CNB-CSIC-IMDEA Nanociencia Associated Unit "Unidad de Nanobiotecnología“
b University of California San Francisco, Mount Zion Cancer Research Center, 2340 Sutter Street, San Francisco, USA
Abstract
Uveal Melanoma (UM) is one of most common tumors of intraocular malignancies. In 90% of the cases, UM is generated due to a single point mutation of GNAQ gene. Currently, the diagnosis of this disease is based on morphological changes of medium-large sized lesions, which are prone to be disseminated to other organs. What is more, the treatment of this metastatic tumor is for now ineffective [1]. Therefore, the development of systems allowing an early detection and treatments of UMs could improve greatly the survival of the patients.
Spherical nucleic acid conjugates (densely oligonucleotide functionalized AuNPs), are nanostructures that present remarkable properties for biomedical applications such as high colloidal stability and biocompatibility. In addition, they show an excellent cellular uptake and due to the negative charged shell, the oligonucleotides are largely protected from nuclease degradation. These properties make them ideal systems to develop biosensors and delivery systems of drugs and oligonucleotides [2].
In this study, we report a single point mutation gene sensor using AuNPs modified with fluorescent Molecular Beacons (Figure 1a). Molecular Beacons were designed to target specifically the mutated GNAQ gene, which can be visualized due to an increase in the fluorescent signal. In addition, we have developed a new release system of siRNAs from AuNPs (Figure 1b), which have shown excellent efficiency to downregulate the mutated GNAQ gene and decrease the cellular viability.
Results showed in this work, evidenced that spherical nucleic acid are excellent new tools to detect the presence of mutated GNAQ gene and modulate its expression [3].
References
[1] Van Raamsdonk, C. D. et al. N Engl J Med, 23 (2010), 2191. [2] Joshua I. et al. J. Amer. Chem. Soc., 134 (2012), 1376. [3] Posch, C. et al. Biomed Microdevices, (2015), 17:15, 14.
Figure 1: a) Schematic structures of AuNPs modified with fluorescent Molecular Beacon. b) New
approach of delivery system of siRNA targeting GNAQ gene. Acknowledgments: IMDEA Nanociencia and Asociación Española Contra el Cancer (AECC) are acknowledged for financial support.
a)
AuNP
5’
3’5’
3’
siRNAAuNP
Molecular Beacon
b)
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venueIMDEA NanocienciaCampus Excelencia UAM-CSIC de Cantoblanco
Calle Faraday,9
28049 Madrid
Spain
how to arrive
Public transport
From Madrid downtown:• Train: C4A y C4B Local (Atocha, Nuevos
Ministerios, ó Chamartín). Cantoblanco
Universidad. Station
• Bus: Line 714 (from Plaza de Castilla).
From Airport:• Underground: Line C8 (to Nuevos Minis-
terios) y then Line C4A y C4B Local (Ato-
cha, Nuevos Ministerios, ó Chamartín).
Cantoblanco Universidad. Station
• Bus: Intercity services: Lines 827/828
(T4); Urban services: Line 204 (T4)
and Line 200 (T1, T2, T3) to Avenida
de América.
By Car
From Madrid downtown:30 minutes. We recommend that you use the
M-30 Nord sens, following signs for Colme-
nar Viejo (M-607), that will guide you to the
University Campus. Take exit 15 (Valdelatas)
and go straight up to coming to the Campus y
then follow the signs to IMDEA Nanociencia.
From Airport:20 minutes. We recommend that you use the
M-40 Nord sens, following signs for Colme-
nar Viejo (M-607), that will guide you to the
University Campus. Take exit 15 (Valdelatas)
and go straight up to coming to the Campus y
then follow the signs to IMDEA Nanociencia.
2015 workshop
final multifun
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