Exploratory research project funded by CNCSIS program PN

Exploratory research project, funded by CNCSIS, program PN II ‘Idei’ THE STRUCTURE OF BIOCOMPATIBLE SYSTEMS BASED ON CORESHELL COLLOIDAL NANOPARTICLES STUDIED BY MOLECULAR PROBES TECHNIQUE Director: Dr. Marilena Vasilescu Project nr. 1317/2009 Lenght: 2009 -2011

Summary The project is oriented towards a very modern research direction – core-shell polymeric nanoparticles – and aims (a) to promote high-level research, (b) to support junior researchers (phd students) wishing to specialize in multidisciplinary fields and (c) to increase the impact of romanian research in the international scientific community. The project's main objective is applying - the molecular probes methods and Uv-Vis, fluorescence, and ESR spectroscopy – in the local structural characterization of nanoparticles concerning their mocropolarity, microviscosity, shape and size, accessibility and the effect of certain additives (medical active substances). The project is articulated along two research directions in copolymers aggregates study: (1) nonionic block coplymers nanoparticles; (2) colloidal nanoparticles based on grafted polyelectrolitic copolymers. The project proposal is based on the experience of a research team with a confirmed competence in complementary fields (chemistry-physics-biology) as well as the efficient employmet of material resources available both locally and at partner institutions (Universtities in Greece and Finland involved in ongoing collaborations). The research activity (including Ph. D specializations) aims at providing fundamental contributions to the nanomaterials research with potential applications in many important fields (medicine, pharmacology, etc. ). Expected dissemination results include publishing articles in high-impact factors publications (ISI), presentations at prestigious scientific manifestations as well as organising scientific manifestations.

2011 TOTAL RON 90000. 00 204000. 00 345996. 00 14443. 30 25000. 00 55500. 00 94943. 3 Mobilities 3252. 16 600 12000. 00 15852. 16 4 Logistics 16968. 36 34400. 00 68500. 00 119868. 36 5 Total Nr. Budget chapters crt 2009 2010 1 Personnel 51996. 00 2 Indirect costs 3 86659. 82 150000. 00 340000. 00 576659. 82

Objectives: The main objective of the project is obtaining polymeric nanoparticles, the study of their microstructure and the study of processes that take place at the level of molecular interactions using the methodology of molecular probes (absorption, fluorescence and spin). We will thus obtain precious data, essential for the use of these materials. More concretely, the scientific aims of the project include: • Obtaining and characterizing core-shell polimeric nanoaprticles on the basis of tribloc copolymers. • Obtaining and characterizing core-shell polimeric nanoaprticles on the basis of graft polyelectrolytic copolymers. • Studying the influence of diverse nonionic organic additives, of the solvent and of various medium-induced factors (p. H, temperature, ionic strength, etc) on the particles properties. • Using various molecular probes to obtain information on micropolarity, microviscosity in core and in shell. • Derivatising and encapsulating nanoparticles with various compound of chemical and biochemical interest. • Involving the derivatized nanoparticles in various chemical and biochemical processes.

Team: Name Birth date Job Ph. D. Vasilescu Marilena 1942 CS I yes Ionita Petre 1967 CS II yes Bandula Rodica 1948 Reseacher no Angelescu Daniel 1971 CS III yes Latus Alina 1982 Reseacher student Bărățoiu Rodica 1979 Reseacher student

Scientific competence of the project manager, Dr. Marilena Vasilescu 1. The study of self-assembling systems using the spectrofluorometry of fluorescent probes The studied systems were micelles, reverse micelles, lyotropic liquid crystalline, hexagonal and lamellar phases, micro-emulsions, gels. The approaching of this domain conducted to a number of international cooperations with: - -University of Helsinki, Department of Physical Chemistry, Helsinki (Finland), Prof. Helge Lemmetyinen 1992 - 1995 “Studies on the aggregation of some surfactants using the fluorescence and spin probe technique “ - University of Uppsala, Department of Physical Chemistry, Uppsala (Sweden), Prof. Mats Almgren, 1992 -2002 : “ Studies on the dynamics and structure of reversed micelles” - -Technical University of Technology (Tampere, Finland) Department of Chemistry , Prof. Helge Lemmetyinen, 1996 - 2003 : “Study of Self-Assembled Surfactant Aggregates, Using Fluorescence and Spin Probe Techniques” - -Technical University of Technology (Tampere, Finland)-Institute of Materials Chemistry, Prof. Helge Lemmetyinen “Dynamics of fast processes: from solutions to self assemblies, studied by spectrophotometric techniques” 2007 -2010 -Department of Chemical Engineering of the University of Patras, (Greece) Prof. Georgios Staikos: “Core-shell colloidal nanoparticles, studied by fluorescence techniques” (2008 -2010). I nvited in Sweden at the University of Uppsala (in the period 1992 -2002) for research visits lasting 2 -3 month, . and in Finland, since 1992, at the Universty of Helsinki and the Technical University of Tampere. I have paid regular visits to the University of Tampere, spending one month on average at each research visit. In 2007 I was invited at the University of Patras, Greece, to give a speech on the topic "Interaction of Globular Proteins and Aluminium Salts with Sodium Dodecyl Sulphate Micelles Studied with Fluorescence Probe Method" 2. Basic research studies in Photophysics. Photoluminescence and Chemiluminescence. 3. New pyrrolic compounds: have been studied both by experimental and theoretical (TD-DFT caculations) methods. With the subject in this domain is the scientific cooperation: “ Structure and Dynamics of Pyrrolic Derivatives in Solutions and Self-assemblies, Studied by Spectroscopic Techniques” betweeb Institute of Physical Chemistry and Technical University of Technology (Tampere, Finland)-Institute of Materials Chemistry, 2004 -2007. Another class of studied compounds was porphyrins. Their study was the subject of CNCSIS grant (2005 -2007). I was the manager of this grant. In the same time the study of porphyrins was the subject of the cooperation with : - Superior Technical Institute, Technical University of Lisbon, Portugal, Prof. Luis Felipe Vieira Ferreira: “Characterization by Advanced Techniques of Tetrapyrrolic Derivatives in Various Environments ”(2005 -2007). Materials: Rare earth oxysulphide phosphors. Medicaments. Semiconductor nanoparticles. Organic-inorganic nanocomposite gels. Composite chromogen-polymeric. Bulk materials and thin films. Organic light-emitting diodes. New materials for light emission and amplification. Photosensitive membranes. Anorganic luminophors.

Dr. Marilena Vasilescu ISI journals published articles (selection from the last 5 years) 1. P. J Aittala, O. Cramariuc, Marilena Vasilescu, R. Bandula, T. I Hukka “Effect of Substituents on the Absorption Properties of Three Pyridylindolizine Derivatives: a DFT and TDDFT Study” Chemical Physics , 360 (1 -3) 162 -170, 2009 , Impact factor 1. 92. 2. Marilena Vasilescu, R. Bandula, O. Cramariuc, H. Lemmetyinen, T. Hukka, F. Dumitrascu “Optical Spectroscopic Characteristics and TDDFT Calculations of New Pyrrolo[1, 2 -b]-pyridazines Derivatives”, J. Photochem. Photobiol. A. , 194, 308 -317, 2008. Impact factor 2. 776. 3. “Synthesis and Characterisation of Europium activated Yttrium Oxide Fine Powders” E. -J. Popovici, L. Muresan, A. Hristea, E. Indrea, Marilena Vasilescu, JOURNAL OF ALLOYS AND COMPOUNDS”, 434 -435, p 809 -812, 2007. Impact factor 1. 61 4. Dan Donescu, Sever Serban, Violeta Uricanu, Michel Duits, Alain Perichaud , Mihaela Olteanu, Manuela Spiroiu, Marilena Vasilescu, ”Filmogen Organic-Inorganic Hybrids Obtained by Sol-Gel in the Presence of cationic Polymer” J. Applied Polymer Science, 106, 26252633, 2007. Impact factor 1. 008. 5. . F. Dumitrascu, M. Caira, M. Vasilescu, L. Barbu, C. Draghici, D. Dumitrescu “Helical chirality of pyrrolo[1, 2 -a][4, 5]diazafluoren-5 -one derivatives”Arkivoc, XVI) 101 -112, 2007. Impact factor 1. 253. 6. M. Bem, F. Badea, C. Draghici, M. T. Caproiu, M. Vasilescu, M. Voicescu, A. Beteringhe, A. Caragheorgheopol, M. Maganu, T. Constantinescu, A. T. Balaban “Synthesis and properties of some new 4 -amino-7 -nitrobenzoxadiazole derivatives”, ARKIVOC, XIII, 87 -104, 2007. Impact factor 1. 253. 7. M. Voicescu, G. Ionita, A. Beteringhe, Marilena Vasilescu, A. Meghea, ”The Antioxidative activity of Riboflavin in presence of Antypirin. Spectroscopic Studies”, J. of Fluorescence, 18, 953 -959, 2008. Impact factor 1. 88. 8. Marilena Vasilescu, R. Bandula, F. Dumitrascu, H. Lemmetyinen“Spectrophotometric Characteristics of New Pyridylindolizine Derivatives Solutions”, J of Fluorescence, 16, 631 -639, 2006. Impact factor 1. 88 9. E. Feitosa, M. R. S. Brazolin, R. R. Naal, Del Lama, Maria Perpetua Freire de Morais; J. R. Lopes, W. Loh, Marilena Vasilescu, “Structural organization of cetyltrimethylammonium sulfate in aqueous solution. The effect of Na 2 SO 4”. J. . Colloid and Interface Science, 299(2), 883889, 2006. Impact factor 2. 755 10. R. Bandula, M. Vasilescu , H. Lemmetyinen “Investigation of the Micropolarity in Reverse Micelles of Nonionic Poly(oxyethylene) Surfactants Using 4 -Nitropyridine-N-Oxid as Absorption Probe”, J. Colloid Interface Sci, 287, 671 -677, 2005. impact factor 2. 755. 11. D. Donescu, A. Banu, S. Serban, M. Vasilescu, M. Olteanu, M. Dudau “Influence of film structure and precursor composition on Rhodamine b retention in dye-dopped ormosils” V. Uricanu, D. , Journal of Sol-Gel Science and Technology, 34, 23 -39, 2005. Impact factor 1. 433 12. M. Vasilescu, F. Dumitrascu, H. Lemmetyinen, N. Tkachenko, ” Steady-State and Time-Resolved Spectroscopic Characteristics of Mesoionic Oxazolones Solutions” Journal of Fluorescence, 14, 443, 2004 Impact factor 2. 038. 13. M. Vasilescu, D. Angelescu, H. Caldararu, M. Almgren, A. Khan “Fluorescence Study on the Size ans Shape of Sodium Dodecyl Sulphate. Aluminium salt Micelles” Colloids and Surfaces A: Physicochem. Eng. Aspects, 235, 57 -64, 2004. Impact factor 2. 097 14 A. Caragheorgheopol, H. Caldararu, M. Vasilescu, Ali Khan, D. Angelescu, N. Zilkova, J. Cejka, ’’Structural Characterization of Micellar Aggregates in Sodium Dodecyl Sulfate/ Aluminum Nitrate/ Urea/ Water System in the Synthesis of Mesoporous Alumina”, J. Phys. Chem. 108 (23), 7735 -7743, 2004 Impact factor 3. 834. .

Dr. Ionita Petre -organic and analytical skills; -synthesis of free stable radicals and diradicals; -generation of short-lived and of persistent free radicals; -electronic paramagnetic resonance (EPR); -spin-trapping of short-lived free radicals; -unconventional nitration, ipso-substitution and mechanistic studies; -spin-labeling and labeling with stable isotopes; -experience with spectro-chemical and physico-chemical analysis; -interphase transport of cations, protons, electrons, and amino-acids using crown compounds; -experimental and theoretical determination of lipophilicities; -chromatographic techniques (TLC, RPTLC); -spin-labeled nanoparticles. List of selected articles 1. P. Ionita, F. Tuna, M. Andruh, T. Constantinescu, A. T. Balaban, Aust. J. Chem. , 2007, 60, 173 -179. 2. P. Ionita, Free Radic. Res. , 2006, 40, 59 -65. 3. P. Ionita, B. C. Gilbert, and V. Chechik, Angew. Chemie Int. Ed. 2005, 44, 3720 -3722. 4. P. Ionita, Chem. Lett. , 2005, 59, 11 -16. 5. P. Ionita, A. Caragheorgheopol, B. C. Gilbert, and V. Chechik, J. Phys. Chem. B, 2005, 109, 3734 -3742. 6. P. Ionita, A. Caragheorgheopol, B. C. Gilbert, and V. Chechik , Langmuir, 2004, 20, 11536 -11544. 7. V. Chechik, P. Ionita, B. C. Gilbert, A. Caragheorgheopol, H. Caldararu, H. Wellsted, A. Korte, Faraday Discuss. , 2004, 125, 279 -91. 8. P. Ionita, B. C. Gilbert, and A. C. Whitwood, Lett. Org. Chem. , 2004, 1, 70 -4. 9. G. Ionita, and P. Ionita, J. Inclusion Phen. , 2003, 45, 79 -82. 10. P. Ionita, A. Caragheorgheopol, V. Chechik, and B. C. Gilbert, J. Am Chem. Soc, 2002, 124, 9048 -9.

CV. Ing. Rodica Bandula a) p. H and solvent effect on the spectroscopic properties of the active pharmaceutical ingredients, surfactants and heterocyclic compunds ; b) Photochemical stabilisation; c) Polymorphysm and molecular conformation of the active pharmaceutical ingredients; d) Self-assembled systems studied by absorption probe technique; structural information regarding the local polarity and viscosity of the selfassembled systems e) Analytical chemstry; List of selected articles “Optical Spectroscopic Characteristics and TD-DFT Calculations of new Pyrrolo [1, 2 -b]pyridazine Derivatives”, M. Vasilescu, Rodica Bandula, O. Cramariuc, T. Hukka, H. Lemmetyinen, Tapio T. Rantala, F. Dumitrascu, J. Photochemistry and Photobiology A: Chemistry, 194, 308 -317 (2008); “Spectrophotometric Characteristics of New Pyridylindolizine Derivatives Solutions”, M. Vasilescu, Rodica Bandula, F. Dumitrascu si H. Lemmetyinen , J. of Fluorescence, 16, 631 (2006); “Investigation of the Micropolarity in Reverse Micelles of Nonionic Poly(oxyethylene) Surfactants Using 4 -Nitropyridine-N-Oxid as Absorption Probe”, Rodica. Bandula, M. Vasilescu si H. Lemmetyinen J. Colloid Interface Sci, 287, 671(2005); ”Spectroscopic Properties of the Fluorescent Mesoionic Oxazolo[3, 2 -b]pyridazin-2 -one in Solution and Solid State”, M. Vasilescu, Rodica Bandula, F. Dumitrascu, C. Draghici si H. Lemmetyinen, Rev. Roum. Chim. , 49(11), 905 (2004); “Molecular Probe Techniques in the Study of Colloidal Systems”, H. Caldararu, A. Caragheorgheopol, M. Vasilescu si Rodica Bandula, Analele Universitatii Bucuresti-Chimie, Anul XII (serie noua), vol. I-II , 173 (2003); “Synthesis of some new solvatochromic 1(4) - substituted Pyrazol-5 -one Azo derivatives”, A. Emandi, I. Serban si Rodica Bandula, Dyes and Pigments, 41, 63 (1999); “Micropolarity and Order in Reverse Micelles of L 62 and L 64 Pluronic Copolymers, as Studied by Molecular Probe Techniques”, M. Vasilescu, A. Caragheorgheopol, H. Cãldãraru, Rodica Bandula, H. Lemmetyinen si H. Joela, J. Phys. Chem. , B, 102 , 7740 (1998); “ Polarity Profiles in Reverse Micelles of Triton X-100 as Studied by Spin Probe and Absorption Probe Techniques”, A. Caragheorgheopol, Rodica Bandula, H. Cãldãraru si H. Joela, J. Molecular Liquids, 72, 105 (1997);

CV Angelescu Daniel Research area of interest - Modeling of the colloidal systems containing oppositely charged macromolecules. - Polyelectrolytes in confined geometries. - Microemulsions as microreactors for synthesis of intermetallic nanoparticles. - Surfactant-polymers at solid-liquid interfaces. - Protein-surfactant and polymer-surfactant systems. - Detergent formulation. - Surfactant assemblies as template for mesoporous inorganic material List of selected articles 1. Jausovec, Darja; Angelescu, Daniel; Voncina, Bojana; Nylander, Tommy; Lindman, Björn, “The antimicrobial reagent role on the degradation of model cellulose film” Journal of Colloid and Interface Science, (2008), 327, 75 -83. 2. Angelescu, Daniel; Linse, Per, “Viruses as supramolecular self-assemblies: modelling of capsid formation and genome packaging” Soft Matter (2008), 4, 1981 -1990. 3. Angelescu, Daniel; Linse, Per, “Modelling of icosahedral viruses” Current Opinion in Colloid and Interface Science (2008), 13, 389 -394. 4. Angelescu, Daniel; Bruinsma, Robijn; Linse, Per “Monte Carlo simulations of polyelectrolytes inside viral capsids” Physical Review E (2006), 73, 041921. 5. Angelescu, Daniel; Caldararu, Horia; Khan, Ali, ”Some observations on the effect of the trivalent counterion Al 3+ to the self-assembly of sodium dodecyl sulfate in water” Colloids and Surfaces, A: Physicochemical and Engineering Aspects (2004), 245(1 -3), 49 -60.

CV Latus Alina Research area of interest Cyclic voltammetry. IR spectroscopy, fluorescence of the active pharmaceutical ingredients. List of Articles Alina Latus, Elena Volanschi, Electrochemical and spectral study of the redox behavior and cardiotoxicity of anticancer drugs with antraquinone structure: quinizarin, Rev. Roum. Chim. , 2008, 53(12), 1131 -1140.

CV Rodica Daniela Baratoiu Research area of interest - Physcochemical characterization of the heterocyclic compounds by UV-Vis, NMR spectroscopy and cyclic voltammetry; - Generation of free radical species with short lifetime via intherphase processes using crown ethers. Published Articles 1. Ba ra t oiu, R. D. , Barbu, A. E. , Mutihac, L. , Ca proiu, M. T. , Dra ghici, C. , Socoteanu, R. , Constantmescu, T. “ 4 -(3 -(Phtalhydrazide)-azo)N-phenylaza-15 -crown-5. Synthesis and properties, ” Revue Roumaine de Chimie 51 (4), pp. 261 -267 2006 2. R. D. Baratoiu, R. Socoteanu, R. C. Mutihac, A. E. Barbu, A. Beteringhe, H. -J. Bushmann, E. Cleve, L. Mutihac, T. Constantinescu and A. T. Balaban Chromo-ionophoric properties of-(5 -phtalhydrazide-azo)phenyle-N-aza-15 -crown-5 in the presence of lithium and sodium ions, Rev. Chim. , 2008, 59, nr. 10, 10731076; 3, Baratoiu, R. D. , Mutihac, L. , Caproiu, M. T. , Draghici, C. , Dumitrascu, F. , Socoteanu, R. , Beteringhe A. . , Manganu M. , Covaci I. C. , Bem M. , Constantinescu T. , Balaban A. T. “ 5 -Dinitro-N-(4’-benzo-15 -crown-5)benzamide derivatives. Synthesis and properties”, Arkivoc, 2008, (xi), 307 -321. 4. R. D. Baratoiu, L. Mutihac, M. Vasilescu, M. Voicescu, A. Latus, G. Ionita, A. Beteringhe, T. Constantinescu, A. T. Balaban, “Chemiluminiscence of para-(5 -phenylene E-azo)-phthalhydrazide-N-aza-15 crown-5”, Arkivoc, in press

Dissemination of the results 1. 05. 2009 -1. 12. 2010 Published or send for publication papers 1. M. Vasilescu, R. Bandula, H. Lemmetyinen, Micropolarity and microviscosity of Pluronic L 62 and L 64 core-shell aggregates in water at various concentrations and additives examined by absorption and fluorescence probes, Colloid Polymer Sci. , 288, 1173. 2010 F. I. 2, 057 2. D. Angelescu, M. Vasilescu, R. Somoghi, D. Donescu, V. S. Teodorescu, Kinetics and optical properties of the silver nanoparticles in aqueous L 64 block copolymer solutions, Colloids and Surfaces A: Physicochemical and Engineering Aspects, 366, 155 -162, 2010. F. I. 2, 172 3. Rodica Bandula si Marilena Vasilescu „Imidazole Derivatives as Absorption Probes for Pluronics Core-Shell Aggregates micropolarity Investigation”, Journal of Applied Spectroscopy, in press , 2011; F. I. 1, 408 4. Marilena Vasilescu si Rodica Bandula, „ Aggregation of Pluronic F 127 and Polydimethylsiloxane-graft-Polyether Block Copolymers in Water and Microstructure of Aggregates as Evaluated by Molecular Probe Techniques”, Revue Roumaine de Chimie, in press, 2011. F. I. 0, 263 5. Vasile Dinoiu, Marilena Vasilescu, Alina Latus, Manuela Savoiu si Liliana Lungu, „The fluorescence and UV-VIS Spectra of Some Plant Extracts in Ethanol and in Polymeric L 64 Micelles”, Revista de Chimie, in press, 2011; I. F. 0, 552 5. Marilena Vasilescu, Alina Latus, Rodica D. Baratoiu, Marioara Bem si Titus Constantinescu, „Pluronic Copolymer Nanostructure for Solubilization of Hidrophobic Benzofuran Derivatives in Water. A Spectrophotometric Study”, Revue Roumaine de Chimie, submitted, I. F. 0, 263.

Dissemination of the results 1. 05. 2009 -1. 12. 2010 Poster presentation at internatonal conference 1. Rodica Bandula, Marilena Vasilescu, H. Lemmetyinen “A Comparative Study by Molecular Probes Techniques of the Pluronics L 62 and L 64 Core-Shell Aggregates Microstructure in Various Environments”, The 16 th Romanian International Conference on Chemistry and Chemical Engineering (RICCCE XVI), 9 -12 September 2009, Sinaia, Romania. 2. Daniel Angelescu si Marilena Vasilescu, „Photophysical Characterization of Pluronic L 64 Triblock Copolymer – Stabilized Silver Nanoparticles in Water”, The 16 th Romanian International Conference on Chemistry and Chemical Engineering (RICCCE XVI), 9 -12 September 2009, Sinaia, Romania. 3. M. Vasilescu, R. Bandula, D. Angelescu, G. Staikos, “Micropolarity and Microviscosity of Core-Shell Aggregates Based on Cationic Comb-Type Polyelectrolyte Examined by Absorption and Fluorescence Probes”, p. 42 Abstracts, The 14 th International Conferene of Physical Chemistry, 9 -12 june 2010, Bucuresti, Romania 4. Marilena Vasilescu, Rodica Bandula si Alina Latus, “Nitronyl Nitroxide Biradicals as Absorption Probes for Core-Shell Aggregates Micropolarity Investigation”, The 7 th International Conference of the Chemical Societies of the South. Eastern European Countries ( ICOSECS 7), 15 -17 September 2010, Bucuresti, Romania. 5. Daniel Angelescu si Marilena Vasilescu “Synthesis, optical properties and Monte Carlo simulations of Cd. S nanoparticles in non-ionic surfactant” p 72 in vol. Abstracts , The 14 th International Conferene of Physical Chemistry, 9 -12 june 2010, Bucuresti, Romania