Project financed by European Commission, ERA-NET Co fund, 7^th Joint Call – 2016

Project title

Translational proof of concept of spintronic micro-nanoparticle non thermal vibrational
therapy for glioblastomas – acronym NANOVIBER

Total project costs: 1,201,273 euro
Total requested budget: 811,530 euro
Total INCDMNR UEFISCDI co-financing: 184,750 euro
Poject duration (months): 36

Link to NANOVIBER Project site

Partners

1. Institut National de la Santé et de la Recherche Médicale (Inserm) Unit1205, Franța, Coordonator

2. COMMISSARIAT A L’ENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES, Franța, Partener

3. Faculdade de Farmácia da Universidade de Lisboa (FFUL) / iMed.ULisboa, Portugalia, Partener

4. INSTITUTUL NATIONAL DE CERCETARE-DEZVOLTARE PENTRU METALE NEFEROASE şi RARE, România, Partener
   Responsabil: Dr. Roxana Mioara PITICESCU, tel: 0213522046, fax: 0213522049, email: roxana.piticescu@imnr.ro

5. ARISTOTLE UNIVERSITY OF THESSALONIKI, LTFN LABORATORY, Grecia, Partener

 Project abstract

Despite the progress of chemo-­‐radiotherapy, molecular and cellular therapies as well as neuro-surgery, glioblastoma remains a deadliness disease. Beside drugs, radiotherapy and surgery, “physical” therapies are synergistic opportunities to solve this major biomedical bottleneck. Paramagnetic parti-cles have been extensively used in association with magnetic stimulation to destroy glioblastoma tissues using thermal strategy. Recently, the impact of vibrational non-­‐thermal strategies has been introduced as a new opportunity. In vitro efficacy has been demonstrated by several groups using innovative micro-­‐nano-­‐particles harboring strong anisotropic magnetic properties. Only one recent publication com-forts the therapeutic efficacy at the preclinical level. To the opposite of the thermal strategy, low cost devices are used to induce vibration providing a strong medico-­‐economic transferability of the technology. Grenoble is one of the few international spot working in this field; the group of B. Dieny is internationally recognized in the field of spintronic for electronics. He recently validated the therapeutic interest of specific spintronic particle for anti-­‐tumor vibrational therapy in vitro. He also developed an innovative vibrational device for in vitro and in vivo vibrational stimulation using simple magnets rotating at 20 Hz thus generating a local rotating magnetic field.

The objective of this 3 years project is to boost the preclinical validation and toxicology of these particles, to support more rigorously a large translational program. The association of European lea-ders in the field of translational nanomedicine, nano-­‐ toxicology, nano-­‐regulation, ethics as well as nanofabrication and nano-­‐characterization should pave for a proof clinical proof of concept trial opening new medical and industrial perspectives in the nanomedicine area.

STAGE 1/2017  ESTABLISHMENT OF METHODOLOGIES OF CHEMICAL-STRUCTURAL CHARACTERIZATION OF FUNCTIO-NALIZED NANOPARTICS

Period: 01.05.2017 – 20.12.2017

Activity 1.1 Characterization of UV-VIS, FT-IR, SEM and complex thermal analysis of functionalized nanoparticles (P4-IMNR)

Activity 1.2 Characterization of the dispersibility of functionalized nanoparticles (P4-IMNR)

The objective of Stage I was to establish the methodologies for chemical-structural characterization of the functionalized nanoparticles.
At this stage they were synthesized by the hydrothermal method at high iron nanoparticle pressures. The nanoparticles thus obtained as well as the samples submitted by the partner P2-CEA / INAC were characterized chemically-structural by the following methods: quantitative chemical analysis, X-ray dif-fraction (DRX), UV-VIS spectrophotometry, FT-IR spectroscopy , optical microscopy (MO), electronic scanning microscopy (SEM) coupled with semicantitative analysis by EDS, complex thermal differential scanning (DSC) thermal analysis and distribution of average particle size by DLS.
Nanoparticles based on iron oxides were synthesized at temperatures of 100-200°C and working pressu-res between 20-1000 bars. It has been found that the synthesis pressure influences the type of crystalli-ne iron oxide phase.
The corroboration of the results obtained in the chemical analysis, DRX, UV-VIS, FT-IR, MO, SEM-EDS and DSC, led to the elaboration of a methodology for chemical-structural characterization of iron oxide nanoparticles. From a dispersibility point of view, a method for characterizing the average particle size in a stable iron oxide suspension in a non-toxic dispersion medium has been developed.

STAGE 2/2018  ESTABLISHMENT OF METHODOLOGIES OF CHEMICAL-STRUCTURAL CHARACTERIZATION OF FUNCTIO-NALIZED NANOPARTICS

Period: 21.12.2017 – 20.12.2018

Activity 2.1 Characterization of thermal stability of functionalized nanoparticles (P4-IMNR)

Activity 2.2 Caracterizarea micro-structurala SEM, TEM, DRX si a proprietatilor magnetice ale nanoparticulelor functionalizate (P4-IMNR)

The objective of Stage II was to carry out methodologies for chemical-structural characterization of functionalised nanoparticles.
At this stage, the chemico-structural characterization methodologies of functionalised nanoparticles were performed by the comparative analysis of commercial magnetotech samples transmitted by P2-CEA / INAC with hydrothermal synthesized magnetite samples in IMNR (P4). Also, samples based on hy-drothermal functionalized iron oxides were analyzed. Thermal stability of nanoparticles of hydrothermal synthesized nanoparticles under various experimental conditions has been demonstrated by performing complex thermal analysis: DSC-TG (including heating and cooling cycles) and DSC. The Curie temperatu-re of the iron oxides obtained (hematite and magnetite) was highlighted. By the chemical-structural characterization of magnetite-based nanoparticles it was found that hydrothermal synthesized powders have crystalline sizes between 17-29 nm; crystallizes in a cubic system in the form of magnetite; form agglomerations of the order of hundreds of nm (when suspended) or aggregates of the micron order (observed by SEM analysis). The sample with the smallest polydispersity index, with the cubic and the best magnetic properties, was synthesized at 200°C and 100 atm. Based on the national and British standards on nanomaterial characterization methods and good laboratory practice on thermal analysis, the DLS method and the TEM / HRTEM microscopy, three characterization methods have been develo-ped: DSC-TG, SEM and DRX..

Dissemination activities:

• M. Cursaru, R. M. Piticescu, A.-M. Mocioiu, D. V. Drăguț, A. G. Plăiașu, C. M. Ducu, „Synthesis and characterization of iron oxide nanoparticles for biomedical applications”, 1^st International Conference on Emerging Technologies in Materials Engineering EmergeMAT and 4^th International Workshop on Materials under Extreme Conditions SUPERMAT, 14-16.11.2018, Bucharest, Romania (poster)
• M. Piticescu, L. M. Cursaru, A.-M. Mocioiu, D. V. Dragut, A. G. Plaiasu, C. M. Ducu, „The influence of synthesis parameters on iron oxide nanoparticles”, International Workshop on Advances in Nanomaterials, 17-19.09.2018, Magurele, Romania (oral presentation)
• 2 ISI papers in preparation
• Morel, C. Thébault, C. Naud, H. Joisten, S. Leulmi, C. Iss, M. Morcrette, G. Ortiz, Y. Hou-Broutin, T. Livache, R. Calemczuk, M. Carrière, P. Sabon, I. Joumard, S. Auffret, et B. Dieny “Vibrating Magnetic Particles for Cancer Cells Destruction”, Invited oral presentation BIMS 2018 – Bio Inspired Magnetic Systems, Exeter (UK), July 9-12, 2018.
• Morel, C. Naud, H. Joisten, S. Leulmi, C. Iss, M. Morcrette, G. Ortiz, Yanxia Hou-Broutin, T. Livache, R. Calemzuck, M. Carrière, P. Sabon, I. Joumard, S. Auffret, et B. Dieny “Design of Magnetic Nanoparticles for Magneto-mechanical Cancer Cells Destruction”, Poster, MagMeet 2018 – 12th International Conference on the Scientific and Clinical Applications of Magnetic Carriers, Copenhagen (DEN), May 22-26, 2018.
• Thébault, C. Naud, E. Billiet, H. Joisten, M. Carrière, Y. Hou-Broutin, R. Morel, and B. Dieny “Magnetic particle oscillations triggered by a rotating magnetic field for mechanical treatment of glioblastoma”, Poster, Congrès SF-Nano 2018 Société Française de Nanomédecine, Montpellier (FR), December 3-5, 2018.
• Patent: “Procédé de fabrication d’un fluide biocompatible comportant une poudre de particules magnétique, fluide biocompatible comportant une poudre de particules magnétiques”, R. Morel, B. Dieny, et H. Joisten. French Patent n° 1852971 deposited on 04/05/2018.

STAGE 3/2019  ESTABLISHING METHODOLOGIES FOR CHEMICAL AND STRUCTURAL CHARACTERIZATION OF FUNCTIONALIZED NANOPARTICLES.
VALIDATION AND CAPITALIZATION OF FUNCTIONALIZED NANOPARTICLES

Period: 21.12.2018 – 20.12.2019

Activity 3.1 Elaboration of the complete methodology for chemical-structural characterization of functionalized nanoparticles

Activity 3.2 Validation of characterization methods of functionalized nanoparticles for in vitro test and clinical trials, part 1

The objectives of Stage III consisted of:
– determination and analysis of the chemical and physical-structural characteristics of nanostructured materials based on iron oxides;
– ensuring the conformity of the analytical characterization methods with the product requirements.

In stage 3 of the project, IMNR (P4) developed two complete methodologies for characterizing functionalized iron oxides. Both methodologies provide essential data for the samples to be certified and transmitted for in vitro characterization. The difference between the two methodologies is the price and the speed of the determinations. Methodology 1 is even faster and less expensive.
4 methods for characterizing the samples based on non-functionalized iron oxides were validated. In stage IV of the project, the characterization methods for samples based on functionalized iron oxides will be validated.
Magnetic measurements have shown that samples based on non-functionalized iron oxide are mostly superparamagnetic at room temperature. The samples do not consist of pure magnetite. The highest total magnetization is found in the samples with the smallest particles.

Dissemination activities:

• 1 ISI publication in Nanomaterials (OPEN ACCESS journal):
  Laura Madalina Cursaru,Roxana Mioara Piticescu, Dumitru Valentin Dragut, Ioan Albert Tudor, Victor Kuncser, Nicusor Iacob, Florentin Stoiciu, The influence of synthesis parameters on structural and magnetic properties of iron oxide nanomaterials, Nanomaterials  10(1), 85, 2020, doi:10.3390/nano10010085, FI=4.324.
• 1 participation in an  international conference (poster): Laura Madalina Cursaru, Roxana Mioara Piticescu, Ioan Albert Tudor, Ana Maria Mocioiu, Dumitru Valentin Dragut, THERMAL INVESTIGATION OF IRON OXIDE NANOMATERIALS. THE INFLUENCE OF SYNTHESIS PARAMETERS ON STRUCTURAL PROPERTIES, 2^nd International Conference on Emerging Technologies in Materials Engineering-EmergeMAT, 6-8 November 2019, Bucharest, Romania
• 1 participation  in EUROINVENT – European Exhibition of Creativity and Innovation 11^th Edition, 16-18 May 2019, Palace of Culture, Iasi, Romania, where the project obtained the Silver Medal, and 2 gold medals from ICECHIM Bucharest and the University of Craiova, respectively.

STAGE 4/2020  ESTABLISHMENT OF METHODOLOGIES OF CHEMICAL-STRUCTURAL CHARACTERIZATION OF FUNCTIO-NALIZED NANOPARTICS

Period: 21.12.2019 – 30.04.2020

Activity 4.1.1. Validation of characterization methods of functionalized nanoparticles for in vitro test and clinical tests, part 2

The objectives of Stage IV consisted of:
– determination and analysis of chemical and physical-structural characteristics of nanostructured materials based on iron oxides functionalized with PEI (polyethyleneimine);
– laboratory validation of 2 methodologies of physical-structural characterization.

In stage 4 of the project, the IMNR (P4) validated 2 methods for characterizing nanostructured compounds based on magnetite and PEI, namely: “Semi-quantitative determination of iron content of iron oxide functionalized with polyethylene imine -PEI by SEM-EDS analysis” and respectively: “Determination of the thermal behavior by DSC-TG analysis of iron oxide functionalized with polyethylene imine-PEI ”.
The obtained results will be the basis for the elaboration of protocols (guidelines) for Analysis by SEM-EDS and respectively Thermogravimetric Analysis for hybrid samples based on magnetite and PEI as a complex alternative to chemical analysis.

Dissemination activities:

• 1 ISI publication in Nanomaterials (OPEN ACCESS journal) in collaboration with Partner P2 – CEA/INAC, France:
  L. M. Cursaru, R. M. Piticescu, D.V. Dragut, R. Morel, C. Thébault, M. Carrière, H. Joisten, B. Dieny, One‐Step Soft Chemical Synthesis of Magnetite Nanoparticles under Inert Gas Atmosphere. Magnetic Properties and In Vitro Study, Nanomaterials, 10, 1500, 2020, doi:10.3390/nano10081500, FI= 4.324.