Multifunctional nanostructures for diagnostic and therapeutic of diseases

Resume : The use of heat to reduce tumor mass is very ancient. Nowadays there are many techniques that allow to focalize the heat in very specific body regions resulting in treatments that are more efficient and minimize side effects. Inorganic Nanoparticlesdue to their nanoscale size,which confer unique physical-chemical features enable their use as heat hubs with external activation. At the same time, given the high surface to volume ratio, the surface of such tiny nanoparticles can be functionaled with biomolecules (i.e. proteins, vitamins, small ligands, etc.) enabling tumor targeting, or with polymer shell for carrying drug molecules and release them under controlled stimuli. Here, two examples of functionalized nanoparticles that can be activated by alternating magnetic field or by near infrared light will be reported. I will first focus on our recent progress on iron-based nanoparticles as heat mediators in magnetic hyperthermia and heat mediated drug carriers (Guardia et al, J. Mat. Chem. B, 2014, 2, 4426, Kolosnjaj-Tabi, ACS NANO, 2014, 8, 4268). Later, I will report about chalcogenide-based nanoparticles as plasmonic nanoparticles for photo-ablation and detection based on Cu64radiolabelling (A. Riedinger, JACS., 2015, 137 151454). Acknowledgements: The author acknowledges the ItalianAIRC project (Contract No. 14527), the Cariplo foundation (Contract No. 2013 0865) and the EU-ITN network Mag(net)icFun (PITN-GA-2012-290248)

Resume : Titanate nanotubes (TiONts) are synthetized by a hydrothermal process (Papa et al., 2009). Their uncommon morphology obtained by controlled parameters allows them to be internalized more easily into cells. This permits their use as novel transfection agents (Papa et al., 2013) without inducing cytotoxicity while providing a radiosensitization effect (Mirjolet et al., 2013). These TiONts are combined to a therapeutic molecule which is docetaxel (DTX) widely used for the treatment of prostate cancer. Currently, injected drugs only reach very weakly tumor sites. Thus, high doses relative to the patient?s requirement are administered causing harmful side effects. In the last decade, the development of nanotechnologies has offered a new strategy to vectorize an active substance specifically in diseased cells. In this work, two developments of nanohybrids are presented to fight against prostate cancer cells and a particular attention was paid on the elaboration of functionalized-TiONts nanohybrids. The first approach consists in combining TiONts and DTX by an original pathway. In vitro results on human prostate cancer cell lines and results of in vivo SPECT-CT images will be presented as well as first irradiation tests. In a second one TiONts and gold nanoparticles, which confers a therapeutic effect by hyperthermia while being detectable by X-ray imaging and MRI (Alric et al., 2008), are combined together. In this context, functionalized-TiONts appear as versatile nanovectors. A.-L. Papa et al., J. Phys. Chem. C, 113 (2009) 12682?12689 A.-L. Papa et al., Nanotoxicology, 7, 6 (2013) 1131-1142 C. Mirjolet et al., Radiother. Oncol., 108, 1 (2013) 136-142 C. Alric et al., Gold Bulletin, 41, 2 (2008) 90-97

Resume : Magnetic nanoparticles (MNPs) have attracted considerable attention for magnetic targeting and hyperthermia applications owing to their ability to generate heat when exposed to an alternative mag-netic field (AMF) without penetration depth limit. Novel magnetic materials for therapeutic agents re-lease based on thermosensitive polymers or vesicles have been developed. In this case, the approach was to induce an increase of permeability of the vector by heat dissipation under AMF excitation. Another way to release drug is to use hyperthermia to break bond between superparamagnetic iron oxide nanoparticles and the target in presence of oscillating magnetic fields. Recently, multifunctional ligands linked to iron oxide nanoparticles took benefits from local heating of nanoparticle’s surface to release a fluorophore on demand. Our approach is motivated by these last developments, i.e. to use local heating as the key parameter to trigger drug release. We synthesised an innovative magnetic delivery nanodevice for targeted cancer therapy showing active control over drug release by using hyperthermia effects. Our material, wich combines the drug controlled release ability of non thermosensitive Molecularly Imprinted Polymers (MIP) with magnetic properties of iron oxide nanoparticles, allows the control release of dox-orubicine. Upon AMF exposure, the bonds between the MIP and the doxorubicin are broken and the molecule is released without any significant heating of the medi-um. These materials offer great promise for the doxorubicin release under alternating magnetic field but moreover we think that this approach can be expanded to other polymers or molecules. Using Magnetic molecularly imprinted polymers for drug delivery under AMF is a major advance in the development of multifunctional targeted drug delivery technologies and may become important theranostic tools in nanomedicines for in vitro and in vivo applications.

Resume : The in vivo detection of rare circulating cells using non invasive fluorescence imaging would provide a key tool to study migration of eg. tumoral or immunological cells. Fluorescence detection is however currently limited by a lack of contrast between the small emission of isolated, fast circulating cells and the strong autofluorescence background of the surrounding tissues. We present the development of near infrared emitting quantum dots with long fluorescence lifetime for sensitive time-gated in vivo imaging of circulating cells. These QDs are composed of low toxicity ZnCuInSe/ZnS materials and made biocompatible using a novel multidentate imidazole zwitterionic block copolymer, ensuring their long term intracellular stability. Cells of interest can thus be labeled ex vivo with QDs, injected intravenously and imaged in the near infrared range. Excitation using a pulsed laser coupled to time-gated detection enables the efficient rejection of short lifetime (≈ ns) autofluorescence background and detection of long lifetime (≈ 150 ns) fluorescence from QD-labeled cells. We demonstrate efficient in vivo imaging of single fast-flowing cells in small animals, which opens opportunities for future biological studies.

Resume : Magnetism in structures with dimensions on the few - nanometer scale have been the center of many investigations ranging from topics in spin-torque and spin-injection dynamics and interactions in biomedical applications to the creation of new types of hard, soft or multi-functionalized materials. Such nanostructured “building blocks” offer new exciting possibi¬li-ties to create new artificial materials.In biological and medical technologies magnetic hybrid or functionalized particles find applications in site-targeted therapy, diagnosis, cell separation and water purification. For innovations in electromobility and wind harvesting new lightweight motors and generators are needed which require the design of new types of permanent magnets – again based on magnetic nanoscale building blocks The talk will give an introduction to the state of the art and address the challenges for tailoring the properties of such building blocks. New functionalities can be obtained based on the intelligent combination of magnetic and non-magnetic materials. Work performed within EU- networks SyntOrbMag, REFreePerMag, IMAGINE and DFG,SFB 445. - H. Zabel and M. Farle (eds.) Magnetic Nanostructures, Springer Tracts in Modern Physics 246 (2013) - Muhammad Arshad Kamran et al.,Tunable emission properties by ferromagnetic coupling Mn(II) aggregates in Mn-doped CdS microbelts/nanowires, Nanotechnology 25 (2014) 385201 - Zi-An Li et al., Chemically-ordered decahedral FePt nanocrystals, Phys. Rev. B 89 (2014) 161406(R) - Zi-An Li, et al., Electrostatic doping as a source for robust ferromagnetism at the interface between antiferromagnetic cobalt oxides , Scientific Reports 5 (2015) 7997 - Chr. Kurts, et al. Splenic red pulp macrophages are intrinsically superparamagnetic and contaminate magnetic cell isolates" Scientific Reports (2015) accepted

Resume : The process of reconstruction of pre-fabricated thin films comprising nanosized particles deposited onto flat substrates triggered by immersion into post-treatment solutions at increasing concentration will be presented and discussed. The evolution of the reconstruction process is assessed by measuring the time (t) dependence of the width (W) of the particle analysis histogram, the W parameter being extracted from atomic force microscopy images. Additionally, a physical picture for modeling the film reconstruction, which provides reconstruction time constants associated to single particles and small agglomerates, the key units associated to the process, ranging from 1 to 10 hours is introduced. The nanoparticle-based film reconstruction triggered by an exogenous stimulus, the use of the W versus t data to describe the process and the model picture accounting for the recorded data have not been previously reported. It is envisaged the disclosed reconstruction process is quite general and may take place in thin films fabricated from nanomaterials regardless the shape, size or chemical composition as long as the solution used for post-treatment contains chemical moieties able to bind the nanomaterial’s surface in one end and bridging together the nanosized units in the other end. Emphasis will be devoted to modulation of properties of thin films engineered for applications in nanobiosensing using the surface reconstruction process.

Resume : Stem cell-based therapies hold great promise in providing desirable solutions for diseases that cannot be effectively cured by conventional therapies. To maximize the therapeutic potentials, advanced cell tracking probes are essential to understand the fate of transplanted stem cells without impairing their properties. Herein, we introduce conjugated polymer (CP) nanodots as non-invasive fluorescent trackers with high brightness and low cytotoxicity for tracking of mesenchymal stem cells (MSCs) to reveal their in vivo behaviours. As compared to the most widely used commercial quantum dot tracker, CP nanodots have shown significantly better long-term tracking ability without compromising the features of MSCs in terms of proliferation, migration, differentiation and secretome. Fluorescence imaging of tissue sections from full-thickness skin wound-bearing mice transplanted with CP nanodot-labeled MSCs suggests that paracrine signaling of the MSCs residing in the regenerated dermis is the predominant contribution to promote skin regeneration, accompanied with a small fraction of endothelial differentiation. To the best of our knowledge, this is the first time exogenous cell trackers could be used to clearly visualize the transplanted MSCs, which revealed that they did not undergo epidermal differentiation because the cells were observed to remain in the regenerated dermis without migration to the epidermis. The facile labeling approach and reliable performance using CP nanodots as long-term cell tracker is beneficial in providing valuable insights into the regenerative capacities of therapeutic stem cells, especially those cells with difficulties in transfection to express fluorescent proteins or luciferase.

Resume : Engineering functional muscle tissue is beneficial for treating various debilitating disorders including myopathy, traumatic injury and malignant tumor extraction through regenerative medicine approaches. However, bioengineered muscles are far in their function compared to those of the natural muscle tissues. Here, we utilized microfluidic spinning to fabricate photocrosslinkable gelatin methacryloyl (GelMA) hydrogel fibers for muscle reconstruction with improved functionalities. Furthermore, we investigated whether the application of topographical cues by surface micropatterning and chemical stimulation by soluble agrin, a neural acetylcholine receptor (AChR) cluster-inducing factor, would facilitate formation and maturation of bioengineered skeletal muscle tissue. Upon culturing cells on the patterned fibers, we observed that C2C12 myoblasts aligned compared to non-patterned fibers. Furthermore, myoblasts cultured on the patterned fibers showed improved differentiation and enhanced myotube formation as indicated by increased myotube length in comparison with those cultured on the non-patterned fibers. Moreover, the addition of agrin during differentiation period of C2C12 cells significantly increased the number and size (length) of AChR clusters on myotubes. Interestingly, combination of agrin treatment with micropatterning synergistically improved myotubes maturity as shown by increased length and aspect ratio of myotubes on the agrin treated patterned fibers. Taken together, the combination of topographical cues and agrin treatment showed to improve the maturation of myoblasts on GelMA fibers, which could be beneficial for bioengineering of enhanced muscle tissue for medical applications.

Resume : In this lecture I will focus on advances in the diagnosis and therapy of cancer associated with the use of specific nanomaterials as products newly emerging nanotechnology. Nanotechnology allows as to manipulate matter at the level of individual atoms and produce materials with new properties that differ from their counterparts in the solid state. Recently, products based on nanotechnology help early diagnosis and treatment of diseases difficult to treat by conventional methods. One of the essential characteristics of nanomaterials is the ability to reach certain cancer cells. Compared with conventional medicines, nanotherapeutics are more effective, safer and allow personalization of the treatment. During the lecture we will presented recent advances in the manufacture of nanostructures and nanomaterials, their specific characteristics and their practical application with examples of pioneering research laboratories and our preliminary studyin the therapy of cancer.

Resume : The integration of multiple functionalities in a single object with nanometric size is crucial for the development of efficient tools for nanomedicine applications. In particular, gold nanoparticles (Au NPs) have been intensively exploited as the starting component for the realization of sophisticated nanomedicine agents, where the desired functionalities are conferred by surface conjugation of various substances such as hydrophilic polymers, proteins for selective targeting, organic molecules with specific spectroscopic fingerprints or stimuli-responsive drugs.1 However, the space available on Au NPs surface is limited and only a finite number of the desired new functions can be hosted. We recently investigated a different approach to the design of Au NPs-based nanomedicines, in which new physical properties are added by doping the gold crystalline lattice to obtain an alloy. This approach, which is usually complicated by thermodynamic constraints and synthetic challenges, becomes accessible by laser ablation synthesis in solution (LASiS). LASiS operates out of thermodynamic equilibrium because NPs are obtained by laser ablation of a solid target immersed in a liquid solution.[1] Here we will present some examples of gold-based multifunctional structures and, in particular, we will focus on binary metal alloys in which the plasmonic properties of gold are combined with the magnetic properties of iron.[2] By acting on the composition of these nanoalloys, one can obtain several properties desirable for a nanomedicine agent, such as simultaneous contrast ability for magnetic resonance imaging, x-ray absorption computed tomography and Raman imaging, or superior plasmon absorption for photothermal applications, or even biodegradability. Besides, these alloys retain the surface chemistry of gold, which is fully exploitable for conjugation with thiolated molecules. Overall, the LASiS approach can be exploited for the realization of a library of multifunctional doped Au NPs and other nanoalloys of interest for nanomedicine applications. [1] a. Laser ablation synthesis in solution and size manipulation of noble metal nanoparticles, Phys. Chem. Chem. Phys., 2009, 11, 3805-3821. b. What controls the composition and the structure of nanomaterials generated by laser ablation in liquid solution?; Phys. Chem. Chem. Phys., 2013, 15, 3027-3046. [2] a. Coexistence of plasmonic and magnetic properties in Au89Fe11 nanoalloys, Nanoscale, 2013,5, 5611-5619. b. Strong dependence of surface plasmon resonance and surface enhanced Raman scattering on the composition of Au?Fe nanoalloys, Nanoscale, 2014,6, 1423-1433. c. Magneto-Plasmonic Au-Fe Alloy Nanoparticles Designed for Multimodal SERS-MRI-CT Imaging, Small, 2014, 10, 2476?2486. d. Superior plasmon absorption in iron-doped gold nanoparticles, Nanoscale 2015,7, 8782-8792 e. Laser generated gold nanocorals with broadband plasmon absorption for photothermal applications, Nanoscale, 2015,7, 13702-13714

Resume : The blossoming growth of nanotechnologies has brought challenging innovations in the synthesis of multifunctional nanoparticles (NPs) for medicine, called ?nanotheranostics? (i.e.) combining diagnosis and therapy [1]. Among functional NPs for theranostic applications, superparamagnetic iron oxides (SPION) NPs are of particular interest because they are biodegradable, FDA-approved, and commercially used as T2-contrast agents for magnetic resonance imaging (MRI) [2]. Moreover, a new area of clinical application displaying a rapid expansion today is magnetic hyperthermia (MH) [3]. When exposed to alternating magnetic fields, magnetic NPs release heat locally, which affects the viability of cancer cells. However, the combination of MH with drug delivery in a unique nanoplatform is still an important scientific lock to leverage to envision clinical applications. Further, innovative strongly anchored and biocompatible nanocoatings of SPION NPs loading and releasing high payloads of drugs are crucially needed. In this work, we developed novel hybrid magnetic core-mesoporous silica nanoparticles loaded with antitumoral agents (doxorubicin, DOX), covered by a human serum albumin (HSA) shell which ensures biocompatibility, stealthiness and biodegradability [4]. The synthesis consists first in the formation of a mesoporous silica shell (~3 nm pore size, 20 nm thickness) around SPION NPs (10 nm diameter). DOX is loaded by impregnation into the small pores of the silica shell and the original grafting of isobutyramide (IBAM) binders [5] at the silica shell surface ensures a tight HSA coating and an efficient encapsulation of DOX. Further, incubation of the DOX-loaded hybrid HSA-coated core-shell NPs in protease media mimicking intracellular lysosomes allowed efficient HSA biodegradation and subsequent DOX release demonstrating the efficient bio-responsive property of such theranostic NPs. These new theranostic magnetic hybrid NPs are currently assessed in various biological studies (ca. cell viability/toxicity, cell uptake, intracellular behavior). References: 1. Adv. Drug Delivery Rev. 2010, 62, 1052-1063; Acc. Chem. Res. 2011, 44, 1029-1038; Chem. Soc. Rev. 2012, 41, 2656-26729. 2. Adv. Drug Delivery Rev. 60, 1252?1265, 2008; Acc. Chem. Res. 44,863?874, 2011; Adv. Drug Delivery Rev. 63, 24?46, 2011. 3. Int. J. Hyperther. 2013, 29 (8), 810-818; J. Neurooncol 2011, 103 (2), 317-324. 4. J. Controlled Release, 2008, 132, 171; J. Controlled Release, 2012, 157, 4. 5. ACS Nano 2012, 6, 7584 ; J. Mater. Chem. 2012, 22, 21434; Nanoscale, 2014, 6, 11676-11680.

Resume : Nowadays, cell imaging is a key-technique for diagnosis, especially for cancer diagnosis. However, classical fluorescence imaging techniques are limited by labels photobleaching, cell autofluorescence or limited resolution. Non Linear optical (NLO) cell imaging provides solutions to each of these drawbacks1. However, the achievement of efficient NLO cell imaging requires the development of new optical probes exhibiting NLO properties. In addition, the coupling of NLO emitters with metal nanoparticles allow to exhibit many additional properties like plasmonic properties, Two Photon Excited Luminescence (TPEL) and therapeutic effect using photodynamic and photothermic therapy. In this work, we synthesized plasmonic NLO labels by coupling tuned gold nanoparticles with SHG emitters such as silicon carbide (SiC) or potassium niobate (KNbO3). The resulting nanohybrids exhibit dual nonlinear optical signals (SHG and TPEL), good localization on cell membranes and no toxicity for healthy cells. Thus, they represent an interesting alternative to the existing fluorescent labels. Finally, to obtain efficient targeting of cancerous cells, nanohybrids were functionalized with targeting ligands. On this contribution, we will present the synthesis of these nanohybrids, the resulting cell imaging and toxicity studies on healthy cells. Finally, cancerous cell targeting will be presented.

Resume : In our current society, nanotechnological applications in health care and industry are blooming, In health care, nanomaterials (NMs) have a lot of potential in the field of imaging contrast agents or targeted delivery of therapy. Although a few types of NMs are being explored in clinical trials[1], their translation from lab to the clinic remains very limited, in part owing to the pertaining uncertainties regarding their safety. More pre-clinical research needs to be performed to keep up with the pace with which novel NMs are being developed[2]. The field of nanotoxicology is a recent field which is rapidly developing into a more mature scientific research area. In its current state, the field suffers from several shortcomings such as the lack of standardization, and the need for more robust and rapid screening methods under physiologically relevant conditions. Here, we present some recent developments on evaluating NM toxicity screening through high-content based imaging combined with gene expression. The methods enable us to 1) link NM safety and biomedical relevance[3,4], 2) link NM safety to cellular NM concentrations up to the single cell level[5] and 3) evaluate specific parameters in detail (e.g. NM degradation). Making use of these detailed data, we are then able to design more safe NMs as well as develop manners to exploit their toxicity specifically on the level of tumor cells as a novel alternative anti-cancer therapy. References. 1. Schütz, C. A. et al. Nanomedicine 2013, 449. 2. Nel, A. et al. Acc. Chem. Res. 2013, 607. 3. Manshian, B. et al. Biomaterials 2014, 9941. 4. Manshian, B. et al. ACS Nano 2015, 10431. 5. Manshian, B. et al. Sci. Rep. 2015, 13890.

Resume : Nanoparticles and their control are of great interest from both academic and industrial points of view, with numerous applications in domains such as medicine, catalysis and material sciences. This talk will stress on metal oxide nanoparticles (NPs) and two aspects: their use as multifunctional contrast agents to help in therapy but also their potential nanotoxicity. As multifunctional contrast agents, they can provide a multiple targeting and visualisation of organs or cells with both detectable changes in the MR signal intensity of the target tissue or organ by changing its MR relaxation properties and classical detectable optical signals for example.The development of biocompatible nanoparticles with an external shell of high-spin paramagnetic lanthanide contrast agents like gadolinium chelate, and europium fluorescent probes led to targeted imaging and gene therapy. A second aspect of NPs is their nanotoxicology which also attracted the attention of public & governments worldwide. Scientists are concerned by addressing the special characteristics of NPs in terms of toxicology and more especially assessing their biological effects. There is an urgent need to evaluate the risks of these particles to ensure their safe production, handling, use, and disposal. In particular, the behavior of NPs inside living cells is still an enigma, and no metabolic responses induced by these particles are understood so far. With this respect, we have examined the potential toxicity due to exposure of TiO2 NPs used in sunscreens & cosmetics. We applied an original imaging methodology (Ion Beam Analysis, TEM, & Confocal microscopy) to in vitro and in vivo studies, combining technologies for detection, tracking, and quantifying TiO2 NPs as well as the use of indicators for ion homeostasis, cell metabolism, or cell fate.

Resume : ZnO nanoparticles have been applied in various areas, such as electronics, cosmetics, medicine, and foods. In particular, ZnO nanoparticles have been used as a zinc supplement and can be also utilized for food packaging as an antimicrobial agent. Although many recent studies have investigated cytotoxicity, toxicity, and biokinetics of ZnO nanoparticles, conflicting results have been reported, thus their potential toxicity after long-term exposure still remains to be elucidated. In this study, the toxicity of ZnO nanoparticles was evaluated by transcriptomic and proteomic analysis after repeated dose 14-day oral administration to rats. Furthermore, bulk sized ZnO nanoparticles and ZnCl2 were also administered in the same manner to compare size and ionized effects. Biological interaction between bulk- or nano-sized ZnO and biological matrices as well as in vivo dissolution property was also evaluated to determine biological fate of two different-sized particles. The results showed that nanocorona profiles binding to ZnO and transcriptomic response analyzed by next-generation sequencing were highly dependent on particle size or biological fate, although no significant toxic effects were found after repeated oral administration. These findings will provide critical information about the toxicity mechanism of ZnO nanoparticles and be useful to understand and predict their toxicity potential.

Resume : The occurrence of tumour diseases in both animals and humans is continuously increasing. Research in nanosciences and molecular biology has put lately an intense effort to identify the aetiology factors and seek for new ways of diagnostic and targeted therapies aimed at reducing mortality and increasing chances to healing. Extensive development of cancer tumours is frequently counteracted through surgery. Assessment of a clean surgical margin is vital and a precise and rapid diagnostic down to molecule level represents a technical challenge with important clinical implications. We present a new way of using surgery instruments and surface enhanced Raman spectroscopy for ex vivo and in vivo diagnostic of clean margins in real time. Raman spectroscopy extracts chemical information with reported 100%sensitivity, 100% specificity and overall accuracy of 93% in identifying carcinomas. Use of surface plasmon activity of silver versus gold nanostructures involved in the surgery instrumentation is investigated in terms of intensity and resolution of the Raman signal. Operation of10 patients (cats and dogs) were assisted with a visible (632nm) excitation source for the Raman effect. The margin diagnostic results were compared with histopathology and anatomic pathology analyses. Excellent resolution and no fluorescence background were remarked. Acknowledgment.This work has been supported from PCCA 2013-Contract 20/2014 and the National CORE programme of Romania.

Resume : Conjugating with poly(ethylene glycol) or human serum albumin is often used to increase the circulating half-lives of therapeutic proteins such as interferons, but these conjugation strategies usually lead to a heterogenous mixture of positional isomers or a fusion protein with unacceptably reduced bioactivity and low yield. To address these problems, we report a new and genetically encoded protein delivery platform, elastin-like polypeptide fusion (ELPfusion), to prolong the circulating half-life of a protein drug. We demonstrate proof-of-concept of ELPfusion by genetic fusion of an important therapeutic protein -interferon (IFN)- at its C-terminus to an elastin-like polypeptide (ELP) to form a well-defined IFN-ELP fusion protein with high yield and retained bioactivity. Notably, the circulating half-life of the IFN-ELP protein (8.6 h) was 28-fold longer than that of IFN (0.3 h). In a murine cancer model, the IFN-ELP protein showed a 65-fold increase in tumor accumulation as compared to IFN 6 h post administration. Furthermore, it completely inhibited tumor growth without appreciable toxic effect, while IFN was just slightly effective at the same dosing. These findings show that ELPfusion may provide a new solution to precisely design protein conjugates with high yield, well retained bioactivity and significantly improved pharmacological profiles.

Resume : This lecture will take place in the field of nanomedicine and will focus on a new class of ultrasmall nanoparticles (AGuIX) that can act as contrast agent for MRI and can help to deliver locally in tumors very high doses under irradiation. These nanoparticles made of a polysiloxane core and surrounded by gadolinium chelates are now synthesized with cGMP conditions for further clinical studies. They display high radiosensitizing efficiency in vitro and in vivo even at very small concentrations and for many different tumor models and irradiation types resulting in important increase of the lifespan for the animals irradiated in presence of nanoparticles. The nanoparticles can accumulate passively in tumors after intravenous injection due to enhanced permeability and retention effect. This accumulation can be followed by MRI and the best moment for irradiation can be monitored precisely. After intravenous injection, the nanoparticles are relatively rapidly eliminated by the renal way with no evidence of toxicity as shown by regulatory studies on rats and monkeys. Due to these encouraging results, AGuIX nanoparticles will be used for a phase I clinical assay on brain metastases.

Resume : Carbon nanostructures (CNSs), among which carbon nanotubes, graphene quantum dots, carbon nanodiamonds, play a major role in nanotechnology. Thanks to their electronic and mechanical properties, can be counted among the most promising nanomaterials in the field of molecular electronics, but they are also proving to be systems of interest in biomedicine, such as innovative materials for neuroscience and drug delivery. They can be used as building block for the preparation of new theranostic platforms. In particular the use of these structures is interesting for the possibility of multifunctionalization, both from the qualitative and the quantitative point of view. Many systems of release of anticancer drugs based on this type of nanostructures are studied in an attempt to take advantage of the ability to preferentially accumulate in tumor tissues due to the so-called EPR effect (Enhanced Permeability and Retention). In this presentation, the most significant results in this field will be presented.

Resume : Currently, multifunctional nanoparticles (NPs) have become a major interest in the field of nanomedecine. These NPs may be designed for reducing the diagnosis times of some tumors and thus improving the survival rate of patients by early treatments. In this work, upconverting nanoparticles (UCNPs) based on NaYF4:Yb,Tm system were successfully dendronized for fluorescence medical imaging applications. UCNPs have attracted much attention due to their peculiar properties, relevant for bioimaging applications in the infrared range. These materials permit the conversion of near infrared (NIR) radiations into photons of higher energy (NIR or visible) via a multiphoton mechanism. In comparison with UV excitation, fluorescence imaging based on NIR light, which is only weakly absorbed by biological tissue, leads to deeper and non-invasive diagnosis.The structural, morphological and optical characterizations of resulting core/shell NaYF4:Yb,Tm@dendrons nanoparticles were performed. In vitro cytotoxicity assays have evidenced their low toxicity. In vivo fluorescence imaging study was performed in mice upon IR excitation, showing promising imaging capacities at low concentrations (0.5 mg/mL) and low power (50 mW/cm²).

Resume : The increasing need for sensitive, low-cost and miniaturized DNA detection methods is one of the causes for the convergence of such apparently different disciplines as biochemistry and nanoscience. Traditionally, nanomaterials such as carbon nanotubes have been of primary interest for developing DNA sensors. On the other side, indium tin oxide (ITO) is a very popular material in the field of optoelectronics for its extraordinary electrical and optical properties. The present work is an investigation on the viability of developing DNA sensors based on ITO nanostructured electrodes. For this purpose, ITO was evaporated by electron beam machine onto silicon electrodes as nanostructured films. Then, a chemistry protocol was used to attach single stranded (ss-) DNA probe chains on the electrodes surface. Then, ferrocene-labeled complementary ss-DNA chains were presented to the biosensor, and the hybridization onto the sensor surface permitted their detection by cyclic voltammetry (CV), basing on reduction/oxidation (redox) properties of ferrocene molecules. Besides, the biosensor selectivity was tested by presenting ferrocene-labeled non-complementary ss-DNA chains to the DNA-functionalized surface. In this case, no hybridization was produced and hence no redox peak was detected by CV, which proved the biosensor’s selective affinity for complementary ss-DNA oligocodes.

Resume : Nanomedicine can take advantage of the recent developments in Nanobiotechnology research areas for the creation of platforms with superior drug carrier capabilities, selective responsiveness to the environment, unique contrast enhancement profiles and improved accumulation at the disease site. Colloidal inorganic nanoparticles (NPs) have been attracting considerable interest in biomedicine, from drug and gene delivery to imaging, sensing and diagnostics. It is essential to modify the NPs surface to have enhanced biocompatibility and reach multifunctional systems for the in vitro and in vivo applications, especially in delivering drugs locally and recognizing overexpressed biomolecules. This presentation will describe the rational design for dendrimer-nanoparticle conjugates elaboration and review their state of the art uses as efficient nanomedicine tools. REFERENCES Felder-Flesch D, Billotey C, Begin-Colin S, et al. Biomaterials, 2011, 32, 8562-8573; New J. Chem., invited Review on Dendrimers in Nuclear Medicine, 2012, 36 (2), 310-323; Eur. J. Inorg. Chem., invited Microreview on Contrast Agents, 2012, 1987-2005; Chem. Commun. 2013, 49, 9158-9160; Nanoscale 2013, 5, 4412-4421; J. Mater. Chem. B. 2014, 2, 1779-1790; New J. Chem. invited paper in SUPRABIO Issue, 2014, 38, 5226-5241; Chem. Mater. 2014, DOI: 10.1021/cm5019025; Nanomedicine Future Medicine Invited review on Dendritic tools on Nanomedicine, 2014, will be published in the April 2015 issue.

Resume : Mesoporous silica nanoparticles (MSN) have attracted much attention the last decade for nanomedicine applications due to their biocompatibility, flexible functionalisation, tunable pore size and diameter. We describe here MSN engineered for two-photon triggered drug delivery or photodynamic therapy, in MCF-7 breast cancer cells. The two-photon triggered drug delivery system was based on a FRET mechanism from a two-photon dye in the walls of the MSN to an azobenzene moiety in the pores of the MSN (nanoimpellers). Concerning photodynamic therapy, a two-photon photosensitizer was encapsulated in the walls of the MSN. Two-photon photodynamic therapy was performed in vitro and in vivo on mice bearing colon xenografted tumors. We also studied the use of porous silicon nanoparticles (pSiNP) functionalized with both a photosensitizer and a targeting agent. pSiNP had been shown to be degraded into non-toxic silicic acid byproducts in vivo. The multi-functionalized pSiNP studied here were able to target, image and kill cancer cells in vitro by photodynamic therapy mechanisms both with 1-photon and 2-photon excitation. Alternatively, the synthesis of disulfide-based biodegradable Periodic Mesoporous Organosilica Nanoparticles (nanoPMOs) was realized and the nanoparticles were efficient in delivering doxorubicin in cancer cells.

Resume : Recent progresses in nanotechnology offer exciting opportunities in the development of promising nanomaterials for medicine and bioimaging. Among them, multifunctional nanoparticles offer plenty of applications because of their physical properties (optic, magnetic?) which allow their use as well as contrast agents, as cell labelling tools, as hyperthermia mediators and smart drug delivery systems (DDS). The common point between these nano-objects lies in their surface chemistry which has to be mastered in the aim to control their physico-chemical features in terms of non-specific binding, targeting efficiency, or drug release. This talk will stress on current surface modification strategies of multifunctional nanoparticles developed for biological applications through two topics: - The nano-bio interface engineering using nanoparticles for in vivo imaging. Their small size, similar to that of biological entities, enables the labelling of biomolecules of interest. For instance, these nanomaterials can be used for in vivo active targeting labelling and cell tracking in the frame of diagnostic or therapeutic applications. All synthesis stages have to be controlled from the inorganic cores surface modification to the conjugaison of functional ligands such as antibodies. We experienced different application fields in biology as various as tumour labelling [1], monocyte and stem cells tracking [2,3], diagnosis of the instability of atherosclerotic plaques [4], etc. - Drug Delivery Systems (DDSs) remotely triggered by magnetic induction. In this field, functionalized magnetic nanoparticles used as heat mediators for self-regulated heat release were designed [5]. Another example of DDS based on multifunctional nanoparticles made of a mesoporous matrix allowing the transport of bioactive molecules containing superparamagnetic nanoparticles will be presented [6]. This type of system provides an interesting tool for increasing the efficacy of pharmaceuticals through improved pharmacokinetics and biodistribution. References 1. F. Sonvico, Bioconjugate Chem., 16, 1181 (2005) 2. A.-K. Bouzier-Sore, NMR Biomed. 23, 88 (2010) 3. C. Lalande, Eur. Cells Mater., 21, 341 (2011) 4. M.-J. Jacobin-Vallat, NMR in Biomed., 24, 413 (2011) 5. R. Epherre, J. Mater. Chem. 21, 4393 (2011). 6. J. Liu, J. Mater. Chem B DOI: 10.1039/c3tb21229g.

Resume : Silica based drug delivery systems have emerged as an important field in biomedicine. However, their development is constrained by the tricky and numerous steps for their preparation. Within that context, we aimed at developing simpler methods for preparing biofunctional mesostructured silica nanoparticles. Our strategy relies on the use of PolyIon electrostatic Complex (PIC) micelles, obtained by complexation between a neutral ionisable double-hydrophilic block copolymer (DHBC) and an oppositely charged micellization agent, as structure-directing agents of silica. Once the mesostructured silica nanoparticles are obtained, it is possible to take advantage of the pH sensitivity of the PIC assembly to release the micellization agent. Two different synthetic routes of bio-functional hybrid silica nanomaterials were developed. The first one involves the formation of a hybrid material using oligochitosan (OC) as micellization agent. It is followed by OC extraction in water by pH control, leading to a mesoporous material functionalized with complexing units of the DHBC able to further encapsulate drugs (neomycin, porphyrin derivatives). In the second approach, the micellization agent used is a therapeutic agent (antibiotics), which allows obtaining directly a hybrid mesostructure loaded with the drug, whose amount can be controlled by varying the DHBC block lengths[1]. Subsequently, release of the drugs can be achieved by controlling the pH of the extracting medium. [1] E. Molina et al. Langmuir 2015, 31, 12839

Resume : In cancer research, the number of scientific works searching alternative therapies for chemotherapy and radiotherapy is continuously increasing, due to the severe side effects that are associated to them. In this context photothermal (PTT) and photodynamic therapies (PDT) are interesting for cancer treatment. Upconversion nanoparticles (UCNP) are being extensively studied for this purpose. We present the synthesis and characterization of two systems of core@shell UCNP with interesting spectroscopic properties for use in PDT. UCNP were synthesized by the thermal decomposition method in the pure hexagonal phase. For the NaYbF4@NaYF4 system, the core@shell structure was characterized by TEM as a core composed mainly for the Yb(III) and a shell composed by a mixing between Y(III) and Yb(III). This sample presented the possibility of anchoring with any PS with absorption around 650 nm for PDT and simultaneous imaging in the Visible or NIR region, besides the resulting white emission under 980 nm excitation. The NaGdF4:Pr:Er:Yb@NaYF4 NP were anchored with Rose Bengal molecule that produces 1O2. The emission properties of the NP were adequated for absorption of the RB molecule and the first results of 1O2 production were made using DPBF as singlet oxygen probe. The results show that the association of the two kinds of NP here presented could be a proposal of a system for theranostic applications using only NIR (980 nm) incident radiation.

Resume : mRNA is an important biomolecule due to its involvement in various diseases (e.g. cancer). Thus, lately the detection of messenger RNA (mRNA) using fluorescently labelled gold nanoparticle-oligonucleotides has been under an intense research interest as potential diagnostic tool.1 At the same time, silencing RNAs (siRNAs) have been introduced as an effective biomolecule for early treatment of diseases playing an essential role in reducing the overexpression of key proteins.2 In recent years, the use of gold nanoparticles as carrier vesicles for RNA has been shown to be a promising system for RNA delivery directly related to applications in imaging and multiplex theranostic treatments.3 In this work, we present the application of fluorescently labelled oligonucleotide coated gold nanoparticles in a complex tissue model (ex vivo skin) for the imaging and manipulation of scar tissue, which demonstrates their potential as multiplex theranostic tools. On the one hand, the oligonucleotide–gold nanoparticles sense mRNAs related to the wound healing process and on the other hand, they downregulate genes related to the expression of proteins involved in tissue scarring.

Resume : Recent years, nanomaterials have attracted more and more attention, and are widely applied in bio-field, such as bio-imaging, drug delivery and tissue engineering. In general, there are two kinds of nanoparticles: fluorescence nanoparticle and non-fluorescence nanoparticle. For most fluorescence nanoparticle, they are cytotoxicity, such as CdS, InP and so on. How to prepare biocompatible fluorescence nanoparticles for bioimaging still is a great challenge. (Fluoridated) Hydroxyapatite (FAp/HAp) is a kind of autologous material with high biocompatibility and well biodegradability. By doped with rare earth elements or combined with other inorganic fluorescence materials, FAp/HAp will possess fluorescence with relatively long life compared with common organic fluorescence materials. What?s more, fluorescence FAp/HAp also can be used as drug delivery for cancer treatment, which can be tracked in vivo. The synthetic method of FAp/HAp nanoparticles is mainly based on hydrothermal/solvothermal method, which has been developed for years. However, precisely control of nanoparticles? morphology and size is still a great problem for their practical applications. In addition, in most cases, the excited wavelength of the fluorescent FAP/HAp nanoparticles could not match the light source attached on commercial laser confocal microscopes or other equipment, which restricts their applications in bioimaging. Recently, our group have focused on synthesis and property investigation on codoped FAp nanoparticles with emitted double color at single excited wavelength, and carbon quantum dots assembled-HAp composite nanorods. We applied the nanoparticles into cell imaging and drug delivery. Further, we used these nanoparticles to modulate stem cells fate and promote cells directional differentiation. Through our work, we have gotten some important results, and believe that these multifunctional fluorescent (fluoridated) hydroxyapatite nanoparticles will have very important application in bio-field. In this talk, we will present the progress on multifunctional fluorescent (fluoridated) hydroxyapatite nanoparticles and their applications in bioimaging, drug delivery and effect on differentiation of stem cells.

Resume : Theranostic nanomedicine is capable of diagnosis, therapy and monitoring the delivery and distribution of drug molecules and has received growing interest. Herein, a self-monitored and self-delivered photosensitizer-doped FRET nanoparticle (NP) drug delivery system (DDS) is designed for this purpose. During preparation, a donor/acceptor pair of perylene and 5,10,15,20-tetro (4-pyridyl) porphyrin (H2TPyP) is co-doped into a chemotherapeutic anticancer drug curcumin (Cur) matrix. In the system, Cur works as a chemotherapeutic agent. In the meantime, the green fluorescence of Cur molecules is quenched (OFF) in the form of NPs and can be subsequently recovered (ON) upon release in tumor cells, which enables additional imaging and real-time self-monitoring capabilities. H2TPyP is employed as a photodynamic therapeutic drug, but it also emits efficient NIR fluorescence for diagnosis via FRET from perylene. By exploiting the emission characteristics of these two emitters, the combinatorial drugs provide a real-time dual-fluorescent imaging/tracking system in vitro and in vivo, and this has not been reported before in self-delivered DDS which simultaneously show a high drug loading capacity (77.6%Cur). Overall, our carrier-free DDS is able to achieve chemotherapy (Cur), photodynamic therapy (H2TPyP), real-time self-monitoring of the release and distribution of the nanomedicine (Cur and H2TPyP). More importantly, the as-prepared NPs show high cancer therapeutic efficiency both in vitro and in vivo. We expect that the present real-time self-monitored and self-delivered DDS with multiple-therapeutic and multiple-fluorescent ability will have board applications in future cancer therapy.

Resume : Inorganic nanoparticles have been widely applied to various industrial fields and biological applications. Especially, zinc oxide (ZnO) and silicon dioxide (SiO2) nanoparticles are used in various food products to improve functionality and stability as a nutritional supplement and an anti-caking agent, respectively. However, there is little information about the physicochemical properties of nanoparticles in foods and their interaction with food components or biological matrices, which can eventually affect their biokinetics and in vivo toxicity. In this study, interaction between ZnO or SiO2 nanoparticles and food ingredients or biological matrices was assessed in terms of changes in their physicochemical properties, in vitro and in vivo solubility, and fluorescence quenching to determine biological fate. Quantitative analysis of saccharides, lipids and minerals which interact with nanoparticles was performed using HPLC, GC-MS and ICP-AES respectively. The results demonstrate that nanoparticles can interact with food components or biological matrices, inducing changes in their physicochemical properties in foods or biological fluids and high fluorescence quenching ratio. Biological fate of nanoparticles was highly dependent on their in vivo dissolution property. Further study is needed to investigate whether these interactions will affect absorption efficiency and potential toxicity as well.

Resume : Silicon dioxide and titanium dioxide nanoparticles have been applied to food industry as an anticaking and food coloring agents, respectively. However, there is little information about their undesirable effects on human health and biokinetic behaviors. In this study, physicochemical properties, intestinal transport mechanism, and biokinetics of food additives silicon dioxide and titanium dioxide nanoparticles were evaluated with respect to particle size (nano versus bulk). We also evaluated the effects of the presence of food ingredients, such as saccharides and proteins, on oral absorption. Biokinetic results demonstrated that silicon dioxide nanoparticles rapidly and largely entered the bloodstream as compared to bulk-sized materials after a single-dose oral administration to rats, while oral absorption efficiency of titanium dioxide was not affected by particle size. Both nanoparticles were determined to be more effectively transcyrosed by microfold (M) cells, present in intestinal epithelium, using in vitro model of human follicle-associated epithelium model. No significant effects of particle size of both materials on tissue distribution and excretion were found. On the other hand, oral absorption of both nanoparticles was extremely low and also highly affected by the presence of food ingredients. These findings will be useful to understand and predict potential toxicity of food additives inorganic nanoparticles.

Resume : With increasing attention to healthy foods, nanotechnology have been attempted to develop functional foods and high value agricultural products. Especially, gold nanoparticles (Au-NPs) have shown to have capability to extend shelf-life and enhance sensory characteristics and functionality of some crops, such as rice, apple, and peach. In this study, the toxicity of colloidal Au-NPs and the extract of Au-NPs-treat red ginseng was evaluated in human intestinal INT407 cells and after 14-day repeated oral administration to rat, respectively. As a result, colloidal Au-NPs showed no cytotoxicity in INT-407 cells after 24 h exposure in terms of inhibition of cell proliferation, membrane damage, and generation of reactive oxygen species, although colony forming ability significantly decreased by exposure to Au-NPs for more prolonged time at high concentration. Biokinetic study revealed that extremely small amount of Au-NPs was absorbed into the blood stream and biological fate of Au-NP is primarily in nanoparticulate form at the systemic levels. 14-day repeated oral toxicity study of Au-NPs and the extract of Au-NPs-treated red ginseng demonstrated no significant changes in body weight, mortality, hematological and blood biochemical parameters, and histopathological morphology. Overall results suggest that Au-NPs and Au-NPs-treat red ginseng did not cause severe toxic effects, implying their excellent potential for high value agricultural products.

Resume : Drug delivery systems can successfully recognize the diseased cells and liberate biologically active compounds directly to target. In the present work, silica carriers equipped with molecular and supramolecular pH-responsible nanovalves were designed by combination of sol-gel synthesis and selective post-synthetic modification. Mesoporous structure of synthesized materials was characterized by low temperature nitrogen adsorption-desorption, small-angle X-ray diffraction and transmission electron microscopy. Chemical immobilization of N-[N’-(N’-phenyl)-2-aminophenyl]-aminoalkyl groups was confirmed by IR spectral and chemical analysis of surface layer. Protolytic properties of grafted aromatic amino groups were studied by potentiometric titration method. It was found that basicity of aromatic amino groups of MCM-41-type mesoporous silica surface increases in the presence of β-cyclodextrin. Loading and release behavior of silica drug carriers was studied in phosphate buffer solutions with pH = 7.0 and 5.0, correspondingly, using doxorubicin as a test compound. It was shown (by UV spectroscopy measurements) that the loading efficiency of mesoporous materials reaches 59-76 %, whereas cumulative values of doxorubicin released from silicas with surface molecular and supramolecular nanovalves into the phosphate buffer solutions with pH = 5.0 achieve 48% and 51%, respectively. Obtained results demonstrate that aromatic amino groups and surface supramolecular structures localized near by pore openings play essential role in pH controlled liberation of doxorubicin.

Resume : Injectable, sustained release drug delivery offers the potential for reducing medical complications by improving patient adherence. [1] Solid drug nanoparticles (SDNs) [2] have the highest drug loading of nanomedicines and offer great potential as reservoirs for long-term drug delivery. However, currently there is only no control of the rate of drug release from such depot sites. To address this issue, we have developed a novel polymer system which entraps the SDNs a polymer gel upon injection. We have shown that thermally-responsive behaviour of poly(N-isopropylacrylamide) nanoparticles can be designed to gel under specific conditions. Heating of the PNIPAM nanoparticles above their lower critical solution temperature to 37°C at physiological ionic strength, resulted in the PNIPAM nanoparticles aggregating into a gel. This gel is capable of entrapping up to 40 % w/w of SDNs. Such nanoparticle/gel composites show controlled drug delivery of the HIV drug lopinavir over periods of time >60 days and that the polymer composition can be used to tune the rate of drug release. These nanoparticle/gel composites will offer the potential for sustained release of dissolved drug molecules from an injected depot in the body. Such drug delivery systems may significantly improve patient outcomes for the treatment of long-term conditions such as HIV. References 1. Hoffman, A. S. J. Control. Release 132, 153?163 (2008) 2. McDonald, T. O. et al. Adv. Healthc. Mater. 3, 400?411 (2014)

Resume : In this work, we have developed a new chitosan-nanohydroxyapatite biocomposites with biomedical applications such as tissue engineering and drug delivery. The biocomposites were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), swelling, degradation and biomineralization capability. Drug delivery through the biocomposites was examined using Fosamax (alendronate sodium, a bisphosphonate). The results obtained suggest that the developed biocomposites possess the essential requisites for their application in the fields of tissue engineering and drug delivery.

Resume : SuperParamagnetic Iron Oxide Nanoparticles (SPIONs) are widely used in biomedical applications (hyperthermia, drug delivery and T2 contrast agents for Magnetic Resonance Imaging (MRI)) thanks to their chemical and biological properties [1-3]. This multimodal probe is a great tool for theranostics. For many years, our lab has developed oxide nanoparticles (NPs) with a narrow size distribution by means of a continuous hydrothermal process [4]. These NPs can be coated in situ by organic molecules like catechols, in particular 3,4-dihydroxy-L-phenylalanine (L-DOPA) [4]. This type of ligand leads to an anti-oxidizing effect and an improvement of suspension stability at physiological pH [4,5]. In this study, a multimodal magnetite (Fe3O4) NPs used as contrast agent for MRI/PET (Positron Emission Tomography) double imaging has been developed. This multimodal agent is easily synthesized by the successive adsorption of three nitrodopamine-based PEGs or macrocycle ligands. The grafting of ligands was confirmed by exhaustive characterizations. The imaging agent is stable under physiological conditions and shows no interaction with serum proteins. Yields of radiolabeling indicate an efficient chelation. In vitro, in vivo and biodistribution results on the constituting elements of the final NPs will be presented. Considering these preliminary results, the elaborated nanohybrids appear as promising contrast agents for MRI/PET with a view to cardiovascular diseases imaging. [1] J. Boudon et al., Chem. Commun., 2013, 7394 [2] L. Maurizi et al., Langmuir, 2009, 8857 [3] L. Maurizi et al., J. Biomed. Nanotechnol. 2015, 126 [4] L. Maurizi et al., Chem. Commun., 2011, 11706 [5] G. Thomas et al., RSC Adv., 2015, 78614

Resume : Titanate nanotubes (TNTs) synthesized via hydrothermal process have attracted extensive interest since their first synthesis owing to their special physicochemical properties such as high aspect ratio. So that, they have been used in various applications such as photocatalysis, energy production, medicine (carriers of medicament) [1]. In spite of their special physicochemical properties and their low cytotoxicity [1], titanate nanotubes (TNTs) present some shortcomings which limit their use as biomaterials such as strong aggregation and a poor solubility [2]. To overcome these limits, organic functionalization of TNTs may be a good alternative. This work consists to functionalize TNTs with 3,4- dihydroxycinnamic acid (DHCA), amino acid L-3,4-dihydroxyphenylalanine (L-DOPA), 3-aminopropyltriethoxysilane (APTES), and chitosan in order to make them suitable for a biomedical application. These molecules improve TNTs properties such as long-term stability under physiological conditions (DOPA and DHCA) and good biocompatibility (APTES, chitosan). Experimental conditions (pH, temperature, time reaction, molar ratio…) have been discussed and optimized in order to obtain the highest grafting efficiency for all the studied compounds. The obtained compounds were characterized by a large range of techniques including transmission electron microscopy (TEM), X-ray Photoelectron Spectroscopy (XPS), thermal analysis (TGA), IR and UV-visible spectroscopy, and zeta potential measurements. [1] A-L. Papa et al., Nanotox. 2013, 7, 1131 [2] A-L. Papa et al., Dalton Trans 2015, 44, 739

Resume : Titanate nanotubes (TiONts) and zinc oxide nanoparticles (ZnO NPs) present enormous bio application capabilities. TiONts have already been reported for their application in biosensor, cancer treatment (Mirjolet et al. 2013), bio imaging, bone regeneration and for drug and gene delivery applications (Papa et al. 2013). TiONts possess needle shape morphology with a cavity and open ends. This structure aids in the functionalization and cellular uptake of the TiONts. In the present study cellular interaction of in-house synthesized and characterized TiONts with glial cells were evaluated prior to proposing them as an agent for glioma treatment. These materials were compared for its toxicity, with ZnO NPs known to be toxic to glial cells (Sruthi et al. 2015). In brief, the nanoparticles were synthesized by classical hydrothermal method and the ZnO NPs were synthesised by wet precipitation method. The synthesized particles were characterized by TEM, SEM, XRD, SAED, Zeta potential and surface area measurements, DLS, XPS, IR, and Raman spectroscopies. The particles were analyzed for their biocompatibility and toxicity using glial cell line. MTT assay was carried out to determine the dose response of the particle incubated with the cells. Particle uptake by the cells was determined using TEM and FACS analysis and the cellular vesicles were examined for the presence of particle. The toxicity of nanoparticle is mainly attributed to its ability to generate Reactive Oxygen Species (ROS), measured using DCFH-DA. Cell death induced by the particle and genotoxic potential was studied in detail to better understand the cellular interaction of the particle. The current study will lay a foundation for the safer use of this nanoparticle for the glioma treatment in the future. S. Sruthi et al., Colloid Surface B, 133 (2015) A.-L. Papa et al., Nanotoxicology, 7 (2013) C. Mirjolet et al., Radiother. Oncol., 108 (2013)

Resume : Gallium selenide (GaSe) belongs to the large class of layered semiconductors. It is a promising material for usage in THz electronics, as the basis of various optoelectronic devices, a matrix for hydrogen storage, substrates in planar nanotechnologies, etc. The technologies of GaSe nanotubes, nanowalls, nanowires, nanoparticles with the thickness of only one layer synthesis exist for today. In this communication we discuss a possibility to use GaSe in medicine. As follow from many investigations, porphyrins stored in tumors transform into strong oxidants under high intensity light illumination and can destroy a cancerous growth in some cases. It is well known that tumor tissues contain an excess of water in comparison to normal ones. It is generally accepted that GaSe is water-insoluble. We have found that a contact of GaSe with water (even if water is in a bound state) under daylight illumination at room temperature results in the dissolution of this semiconducting compound and the formation of selenous and selenic acids. The obtained results allow to consider GaSe nanoparticles as a promising material not only for early diagnosis of tumors but also for their therapy.

Resume : As one of the most abundant polysaccharides in nature, amylose is a linear homopolymer of glucose linked with ?(1,4) glycosidic bond. Amylose is known as safe natural material and widely used in food, medicine, and chemical industries with its biocompatible and biodegradable properties. Here, we present a novel approach to prepare pure amylose microparticle by enzymatic synthesis and self-assembly process. Iron oxide nanoparticles can spontaneously be incorporated into the amylose microstructure during enzymatic synthesis to form a well-defined spherical particles with superparamagnetic characteristics. The resulting amylose magnetic particle (AMP) was characterized by TEM, SEM, XRD and VSM magnetometer. For the conjugation of specific antibodies to the surface of AMP, we produced a MBP-SPG chimeric protein by fusing two genes that encode these two proteins and expressing it in E. coli. The specific affinity of MBP and SPG toward amylose and Fc domain of antibody, respectively, enabled us to conjugate antibodies to the surface of AMP. We demonstrated the ability of antibody-conjugated AMP to capture target pathogen, E. coli O157:H7. The target bacteria captured on the surface of AMP were easily concentrated by magnet and be subsequently analysed by quantitative real-time PCR. The antibody-conjugated AMPs were shown to capture over 90% of target bacteria and facilitated the detection procedure by effectively isolating and concentrating low number of target pathogen from a clinical samples.

Resume : Raman spectrometry provides a huge variety of applications in biological and biomedical research. Besides characterization of cells and tissue, it allows for identification and quantification of disease related significant Biomarkers present in human body fluids. Quantitative measurements with the aim to ensure metrological traceability can be performed by applying the isotope dilution (ID) principle. This approach, which results in significantly lower uncertainties, requires the determination of an intensity ratio resulting from Raman signals of the natural biomolecule (analyte) and signals from an isotopologue which has a minimal higher molecular weight than its natural form. In combination with surface-enhanced Raman scattering (SERS), biomarkers at their relevant concentration levels can be measured. Here, we report on an approach that aims in combining the accuracy of the ID-principle with the sensitivity and specificity of a SERS-immunoassay. The preparation of the preferably gold nanoparticles involves coating with a Raman reporter molecule (dye) in a natural and isotope-edited form for detection and quantification and the use of suitable antibodies which are targeted against the biomolecule of interest. The specificity of this coherent complex is given by the antibodies which are used. In this manner, a reference procedure for the quantification of ß-Amyloid and tau-protein in cerebrospinal fluid (CSF) as a diagnostic test for Alzheimer disease shall be realized.

Resume : Tailored nanoparticle properties have wide ranging potential in areas of energy conversion, catalysis, diagnostics, sensing, imaging and therapeutics.[1] Hybrid materials comprising supported nanoparticles are advantageous as they are easy to handle and manipulate, and also provide a highly localized concentration of nanoparticles.[2] An eloquent example of their application is the use of supported particles as radiosensitizers in cells.[3,4] For diagnostic and therapeutic applications the support offers additional avenues for targeted delivery in vivo. Though a variety of methodologies exist to prepare robust composites using gels, polymer networks, micelles, microspheres, achieving controlled coating remains a key challenge. In this study the preparation of a variety of composites will be presented together with the methodologies used to quantify the gold loading, as well as the application of these hybrid materials as radiosensitizers with good dose enhancement factors when exposed to radiation. A variety of families were studied, consisting of polystyrene sphere supports (60 nm - 200 nm) with various sized gold nanoparticles (4.5 nm - 26 nm) immobilized. REFERENCES [1] S. Eustis, M. A. el-Sayed, M. Chem. Soc. Rev. 2006, 35, 209?217. [2] J. M. Campelo, D. Luna, R. Luque, José M. Marinas, A. A. Romero ChemSusChem 2009, 2, 18?45. [3] K. T. Butterworth, S. J McMahon, F. J. Currell, K. M Prise Nanoscale, 2012, 4, 4830-8. [4] S. Jain, J. A Coulter, K. T Butterworth, A. R Hounsell, S. J McMahon, W. B Hyland, M. F Muir, G. R Dickson, K. M Prise, F. J Currell, D. G Hirst, J. M O'Sullivan Radiother Oncol., 2014, 110, 342-7.

Resume : Implantable drug delivery systems (IDDS), which deliver a sustained release of a drug over extended periods of time, have been shown to be effective in several therapeutic areas. [1] If a therapeutic regimen requires a patient to take drug doses frequently or over a long time period, adherence will be impacted leading to increased morbidity and mortality. The preparation of poorly water soluble pharmaceuticals as solid drug nanoparticles (SDN) which can be dispersed in water has significant benefits, [2] including increased oral bioavailability, reduced side effects and improved targeting. [3] We are working on the synthesis of charged polymer microparticles which when combined will form a porous hydrogel network which can store and release SDNs. These charged polymer microparticles consist mainly of hydroxyethyl methacrylate (HEMA) or Dextran-HEMA monomers along with ionisable co-monomers. When oppositely charged microspheres are combined in the presence of SDNs a hydrogel microparticle network will be formed and the release of drug containing nanoparticles from this IDDS will be studied. The advantages of such an IDDS is that it combines the sustained long-term drug delivery with the improved targeting provided by nanomedicines. References 1.Hoffman, A.S., J.Contr.Rel., 2008, 132, (3),153; 2.McDonald,T.O. et al., Adv. Healthcare Mater., 2014, 3, 400; 3.McNeil,S.E., WIREs:Nanomed.Nanobiotech., 2009, 1, 264.

Resume : The tetraruthenium substituted polyoxometalate [Ru4O4(OH)2(H2O)4(gamma-SiW10O36)2]10- (RuPOM) is a nano-sized bio-mimetic inorganic catalyst. We have investigated its catalase (CAT) and superoxide dismutase (SOD) activity in physiological-like environment, demonstrating its potential as robust artificial antioxidant enzyme, for Reactive Oxygen Species (ROS) scavenging. Owing to its polyanionic charge, RuPOM can be easily integrated into nano/micro carriers, by exploiting Layer-by-Layer (LbL) self-assembly procedures. In particular, it was immobilized within the shell of polyelectrolyte multilayer microcapsules (PMCs) containing a soft fluorescent core. Dioxygen bubbles evolved during H2O2 dismutation were shown to propel the movement of the capsules in a aqueous mixture containing a H2O2 gradient. In addition, as highlighted by using a microfluidic channel, the capsules induce an elastic turbulence of the fluid, depending on the kinetic parameters of H2O2 dismutation.

Resume : Breakthroughs in nanotechnology allow the integration of different nanoparticles single hybrid nanostructures (HNSs), forming multifunctional nanosized sensors with an excellent perspective in the life sciences while small luminescent nanoparticles (LNPs) have brought great advances to the areas of imaging and sensing but most of them use ultraviolet or visible light and are hence limited to cellular studies or topical applications. For in vivo bioimaging LNPs excited by and emitting in the infrared hold more promise because of their ability to penetrate biological tissue.[1] Combining our interest in LNPs that emit IR light between 1000 ? 1350 nm, an area with minimal biological tissue absorption and scattering defined as the 2nd biological windows (II-BW) with the strive for multimodal/multifunctional nanoparticles, we developed HNSs based on poly-lactic-co-glycolic acid (PLGA) that include superparamagnetic iron oxide and quantum dots, which absorb and emit in the II-BW. We demosntrated their ability for in vivo multimodal imaging (MRI and infrared fluorescence) in mice, overcoming the tissue penetration limits of classical visible-light techniques. Additionally, their multifunctionality was studied through the assessment of their applicability as potential magnetic heating agents in hyperthermia treatments.[2] In summary, HNSs including magnetic and luminescent nanoparticles were synthesized and applied to in vivo multimodal bioimaging while also demonstrating their magnetic hyperthermia capability. [1] D. Jaque, B. d. Rosal, E. M. Rodríguez, L. M. Maestro, P. Haro-González, J. G. Solé Nanomedicine 2014, 9, 1047. [2] D. H. Ortgies, L. de la Cueva, B. del Rosal, F. Sanz-Rodríguez, N. Fernández, M. C. Iglesias-de la Cruz, G. Salas, D. Cabrera, F. J. Teran, D. Jaque, E. M. Rodríguez ACS Appl. Mater. Interfaces 2016, DOI: 10.1021/acsami.5b10617.

Resume : Non-chemically cross-linked chitosan converted in biogel by sono-chemical synthesis in high density ultrasonic, was evaluated as support for drug delivery aiming to obtain nontoxic functional biopolymer network. The successful crosslinking was performed by OH-radicals generated during cavitation in aqueous solution of acetic acid. The capacity of swelling and pH-responsive, antioxidant activity is estimated for silymarin. In vitro pharmacological tests was performed on 3T3 fibroblasts (ACC 173). The chitosan self -crosslinked network shows a hydrodynamic diameter measured by DLS for chitosan molecules with DDA-80%; a high capacity of swelling up to 96% for silymarin; the antioxidant activity ~ 96% has been measured by chemiluminescence and DPPH methods. Pharmacological evaluation, show significant values for cell viability at application of concentration of 0.1 mg/ mL and 1 mg/ml (96.99% for 0.1 mg/ mL samples and 65.74 - 99.53% for 1 mg/ mL samples). Keywords: drug delivery nano systems, herbal extracts, antioxidant activity, silymarin

Resume : We electropolymerized polypyrrole thin layer onto Au nanowires via cyclic voltammetry (CV) as molecular imprinted polymer (MIP) biosensor to detect dopamine (DA) and catechol (CA). The Au nanowires/polypyrrole core/shell structure were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Raman spectroscopy and linear sweep voltammetry (LSV) was used for sensitivity and selectivity test. Selectivity of MIPs is due to binding sites which are complementary in size and shape to target molecules. Therefore, these sites can recognize targets and rebind them. The DA sensor showed LSV signal intensity at 9.5 μA mM-1 and CA sensor at 10.7 μA mM-1. The determined linear range was 5 x 10-5 - 5 x 10-4 mol L-1 (N=3)

Resume : Early stage of Paramecium bursaria chlorella virus 1 (PBCV-1) infection on Chlorella (green algae) is non-invasively measured using electrochemical field-effect transistor (eFET) in real-time. The eFET made from a network of one-dimensional (1D) Au nanoparticle necklace arrays, which spans over 70 µm, has extreme sensitivity to a single-electron charge at room temperature (RT), resulting in the high specificity and sensitivity of electrophysiological changes of microorganisms. The applicability of the device is demonstrated by measuring the membrane depolarization of the Chlorella and K ion exocytosis from the cells in real-time during the virus infection process. The viral infection is also confirmed by quantitatively measurement of eFET coupled with delivery of viral particles by a scanning probe microscope cantilever.

Resume : Silicon is by far the most important semiconductor material enabling high-performance microelectronic and photovoltaic devices. Application spectrum of silicon is further expanding in the field of optoelectronics and photonics. Recently, significant potential of silicon nanocrystals (SiNC) was revealed for their possible application in biology and medicine. They have been studied especially because of their biocompatibility, low toxicity and natural biodegradability. Such characteristics distinguish SiNC from the current semiconductor fluorescence markers called quantum dots which are inappropriate for the application into the living organism due to their potential toxicity. Nowadays, biomedical research concentrates on the possible use of SiNC as tools for bioimaging, biological sensors as well as for drug delivery. Our specially designed SiNC are co-doped with boron and phosphorus and, from the physical point of view, they display exceptional parameters - well defined concentration and crystal size (3-4 nm) in methanol colloid, long lasting dispersion in aqueous solutions, photostability and photoluminescence insensitive to the changes of the environment and to surface modification. Thus these alloy-like SiNC are applicable in biomedicine. First in vitro experiments with human osteoblastic cell line led to the evaluation of the cytotoxicity of different types of SiNC and to the assessement of the highest non-cytotoxic SiNC concentration suitable for their detection in cells using fluorescence microscopy. Two different types of SiNC (3 and 4 nm) were detected in cells at different time points. Results showed how SiNCs are incorporated into the cells over time. Additionally, we proposed possible influence of the presence or absence of serum (fetal bovine or human) in cultivation media on SiNC cytotoxicity and transport into the cells. Finally, we discuss potential of co-doped SiNC in biomedicine.

Resume : Nanoparticles assemblies present a great interest for the development of new advanced applications such as magneto-plasmonic sensors. The structuration of magnetic nanoparticles in arrays envisioned to produce ultra-sensitive nanosensors of bio-molecules thanks to collective and synergistic properties. Surface plasmon resonance (SPR) of gold substrates functionalized by self-assembled monolayer (SAM) of organic molecules has been modulated significantly by controlling the assembling of iron oxide nanoparticles as a monolayer with different density[1]. On the other hand, ?click? chemistry has been demonstrated to be very efficient to control the structure of nanoparticle assemblies and so to modulate magnetic collective properties[2, 3]. The assembling process is controlled by specific interactions between functional groups located at the surface of both SAM and nanoparticles. In this study, we take advantage of this approach to prepare biosensors based on magneto-plasmonic properties. Iron oxide nanoparticle monolayers assembled, which still have functional groups, are post-functionalized using ?click? chemistry by biomolecules which acts as detectors. The very well-known biotin/streptavidin couple has been investigated to assess the efficiency of such sensors[3]. The surface of nanoparticle layer is functionalized by biotin groups which enable the detection of streptavidin by bio-conjugation with. The adsorption of streptavidin is demonstrated by monitoring the shift of SPR signal from gold substrate as function of reaction time and streptavidin concentration. [1] Pichon, B. P.; Barbillon, G.; Marie, P.; Pauly, M.; Begin-Colin, S., Nanoscale, (2011) 3, 4696. [2] Toulemon, D.; Pichon, B. P.; Cattoen, X.; Man, M. W.; Begin-Colin, S., Chem Commun (Camb), (2011) 47, 11954. [3] Barbillon, G.; Bijeon, J. L.; Plain, J.; Royer, P., Thin Solid Films, (2009) 517, 2997.

Resume : There are many advantages in applying nanotechnology to molecular diagnostics to improve the simplicity and the sensitivity of the analysis. Semi-conductor nanocrystals or quantum dots (QDs) in particular demonstrate several properties that make them good candidates for biomolecular recognition. These nanocrystals present narrow size-tunable emission spectra and a broad excitation spectrum. In addition, they offer higher photostability and brigthness compared to conventional organic dyes. The objective of this study is to develop a new diagnostic tool based on fluorescent nanobeads containing quantum dots for a fast and single-molecule detection. Our strategy is to self-assemble QDs into clusters of a few hundreds of nanometers in diameter for an even brighter fluorescence and easily detectable analytical signal. In order to protect their colloidal and photo-stability, the clusters need to be covered with ligands such as a polymer or a silica shell. Our work focuses on the formation of monodisperse QD-based superparticles (SPs). In order to obtain very stable SPs, we chose to grow a silica shell on their surface. Biocompatibility is further assured by a new type of polymer-silica hybrid presenting a zwitterionic chain for water solubility and reactive functions for conjugation with biomolecules. Preliminary results have shown that we can integrate the SPs into a microfluidic system for an efficient single-particle counting.

Resume : Nanomedicine is one of the most interesting field of application for nanoparticles (NP). In cancer research, many scientists are currently focusing on building multi-purpose nano or micro systems, often obtained by combining NPs with different properties like gold and iron oxide NPs. Gold NPs are of interest in nanomedicine because of their biocompatibility, easy functionalization and optical properties which make them a powerful cells detection tool using, for example, SERS spectroscopy and for hyperthermal treatment of cancer cells. Iron oxide NPs in particular those made of magnetite, can be used for cell sorting exploiting their magnetic properties. We report the synthesis, by using NPs obtained by Laser ablation Synthesis in Solvents, of core-shell-satellite (CSS) sub-micron systems made of an iron oxide NPs core, dye-doped silica shell and AuNPs as satellite decoration. CSS have been fully characterized and show high SERS activity. They are functionalized with an antibody specific for PSMA antigen overexpressed by prostatic cancer cells. We show that antibody functionalized CSS can be efficiently used in the selective immunomagnetic sorting of prostatic cancer cells and, coupling immunomagnetic sorting with single-cell SERS analysis, a reduction of false positive arising from immunomagnetic sorting procedure is obtained. We also show application of CSS in photothermal treatment of prostatic cancer cells using a continuous laser at 647 nm.

Resume : Atherosclerosis is a chronic disease characterized by the formation of plaque in regions of arteries. Current imaging techniques lack specificity for the plaques and consequently, new selective contrast agent is necessary for accurate imaging of plaque. Poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) are appealing Drug Delivery Vectors, since they have the ability to be surface engineered in order to increase the body circulation, direct to specific organs or uptake via targeted cells. Since drug delivery reach the cell interior, is fundamental to understand the uptake of the NPs into cells, quantify it and determine the localization intracellular. In this study, we proceed towards the synthesis of PLGA NPs with encapsulated iron oxide NPs, labelled with quantum dots in the particle core for targeting atherosclerotic lesions. PLGA NPs were loaded with rhodamine 6G, an FDA-approved fluorescent agent and used for intracellular localization studies, applying flow cytometry (FACS) and Confocal Scanning Laser Microscopy (CLSM). Studies of PLGA NPs biodistribution with different surface functionalization, took place, allows us to quantify the amount of NPs per organ and the distribution patterns after intravenous delivery. Cytotoxicity studies with hepatocytes further reinforced the ability of NPs to enhance cellular uptake using FACS-CLSM analysis for cellular localization and quantification. Radiolabelled PLGA NPs are appeared to be an excellent biocompatible model for the evaluation of targeting capacity of nanocarriers, with unique biodistribution, cellular uptake and targeting properties, thus to enhance a prototype platform for drug delivery system, especially for cardiovascular diseases.

Resume : Recently, metal nanostructures are frequently considered for highly sensitive detectors that show potential in biochemistry and medical diagnostics. In this work we report on preparation and characteristics of ultrathin (5-10 nm) Au films deposited onto highly ordered structured titanium templates for SERS (Surface Enhanced Raman Spectroscopy) and electrochemistry applications. The templates are formed by chemical etching of TiO2 nanotubes out of titanium substrate. The surface of the obtained material perfectly maps the bottom layer of the nanotubes and consists of a uniformly distributed dimples with diameter in the range of 30-100 nm. For the structures covered with ultra thin Au films the measured average SERS signal is markedly higher than observed for bare Ti templates. This is due to strong electromagnetic field in the vicinity of the film discontinuities. The electrochemical response of the prepared Au/Ti substrates studied in the presence of glucose solution shows that the current registered at the oxidation peak rises with the increasing glucose concentration on the contrary to bare Ti for which no activity can be observed. The effect depends linearly on glucose concentrations in the range of 0.1 - 40 mM that covers the physiological detection needs regarding the blood sugar. KG and KS acknowledge the National Science Centre of Poland for financial support via grants 2012/07/N/ST5/02139 and 2012/07/D/ST5/02269.

Resume : Localised drug delivery to tumours may be applied by the use of a blood compatible nanosized carrier agent able to respond to an external stimulus (such as a temperature increase) with a ?triggered? drug release. Conventional thermosenstive liposomes (TSLs) or similar delivery systems lack the labels for in vivo tracking or tumour imaging and hence the ability to assess the optimal trigger time post administration. Here we discuss a dual labelled thermosensitive liposomal delivery system for localised delivery by Focussed Ultrasound (FUS) assisted mild hyperthermia. In addition to labelling for MRI, we introduced a Near Infrared fluorescence (NIRF) label which greatly assisted in the near real time tracking of the carrier in our murine xenograft cancer model. This in turn allowed for optimisation of the FUS conditions and timings, allowing for triggered-release and functional delivery of the therapeutic drugs to the tumours. NIRF allowed for high sensitivity imaging with little intrinsic tissue background fluorescence and good transmission & tissue scattering characteristics compared to visible region fluorophores. We synthesised these labels as lipid attached conjugates to ensure specific and lasting labelling of the carrier liposomes. MRI contrast enhancement ability and NIRF signals were assessed in vitro and in vivo. Nanoparticle kinetics in murine tumours were assessed with optical imaging and at defined time intervals post i.v. injection, FUS was applied to induce a small increase in temperature to 42-43°C for 3-5 min. Imaging revealed both dramatic nanoparticles accumulation (5-10 fold) and drug release immediately after induced hyperthermia and only in the FUS treated tumour. Significant tumour growth inhibition was observed for the FUS treated tumours compared to those that were treated only with the drug nanoparticles. The applications of such multifunctional ?nanotheranostics? with short, mild hyperthermia could have a transformative effect on cancer chemotherapy.

Resume : Polysaccharide-based nanoparticles exhibit a great potential as holders for imaging and drug agents because of their intrinsic physicochemical properties being biocompatible and biodegradable. Moreover, they are highly stable, safe, non-toxic, hydrophilic and offer a large variety of functional groups, degree of branching (linear to highly branched) and molecular weight (low to high). Thus, they have a wide variety of composition and properties that cannot be replicated in a chemical laboratory, and the ease of their production makes various polysaccharides cheaper than synthetic polymers. Therefore, the use of polysaccharides as biomaterials is quite promising for biomedical applications. In this work, we will discuss about polysaccharides being a key component in the development of contrast agents for the new magnetic resonance imaging technologies. In this context, different processing strategies for synthesizing different polysaccharide-based composite nanoparticles will be presented. Acknowledgment: This research is supported by the H2020-EU.1.2.1.-founded project ?CONQUER: Contrast by quadrupole enhanced relexation?, Project reference 665172

Resume : Nanoparticles only can be loaded with therapeutic and imaging agents enabling de facto combination therapy and multimodal imaging. In this work, theranostic polymeric nanoconstructs (TPNs) are realized with a core of poly(lactic-co-glycolic acid) (PLGA) stabilized by a lipid monolayer decorated with polypropylene glycol chains (PEG2000) resulting in ~ 200 nm spheres. TPNs are loaded with potent anti-cancer (docetaxel - DTXL) and anti-inflammatory (diclofenac - DICL) molecules for the treatment of brain tumors. The lipid monolayer is further labeled with near infra-red dye (Cy5) and gadolinium-based macromolecules (Gd-DOTA) for biodistribution analysis and therapy follow-up. In vitro, DTXL:DICL TPNs induce tumor cell death two times more efficiently than DTXL-only TPNs, achieving an IC50 of 90.5 nM at 72h on U87-MG cells. In mice bearing glioblastoma multiforme tumors, TPNs demonstrate the highest efficacy: at 70 days post treatment, survival rates are 70%, 40% and 0% for mice treated with DTXL:DICL TPNs; DTXL-only TPNs and free DTXL, respectively. TPNs accumulation within malignant masses of different sizes, namely of ~ 300, 600 and 1,000 mg, is studied via both fluorescent optical imaging (IVIS Spectrum) and elemental analysis for gadolinium via inductively coupled plasma mass spectroscopy (ICP-MS). Longitudinal relaxivities as high as 60 (mM·s)-1 are documented at 1.41T resulting in significant contrast enhancement in 3T clinical Magnetic Resonance Imaging (MRI) scanners.

Resume : Prostate cancer is the second cause of cancer in male population, with the risk of cancer developing typically increasing with age and with familiarity. Early stage detection is crucial for a positive prognosis of the disease when it is still localized within the capsule, before its metastatic transformation. The development of novel and non-invasive approaches for the detection and control of prostate cancer progression is increasing in importance because they could give the possibility of a wide population screening. Nowadays non-invasive detection methods are mainly based of the detection of specific tumor markers inside urine, but unfortunately they are neither enough sensitive, neither enough specific. Prostate-specific antigen (PSA) is an FDA approved marker for prostate cancer screening in USA. Levels between 4 and 10 ng/ml in the blood are considered to be suspicious and prostate biopsy is performed to confirm diagnosis. Laboratory techniques for PSA detection are often expensive, time consuming and they require specialized technicians. In our laboratory, we are developing an Organic electrochemical transistor (OECT) based on PSA antigen incorporated in a PEDOT:PPS conductive polymer for PSA recognition in the urine. This OECT should detect PSA level around 4 ng/ml or lower, it is reusable, economic, non-invasive and easy to use thanks to the embedded electronics. Thus, when optimized, it can be a valuable candidate as a tool for extensive prostate cancer screening.

Resume : In the present work, we have synthesized Fe3O4 magnetite ferrite nanoparticles using coprecipitation method, in this study we have controlled the PH of the reaction as it is an important parameter during synthesis. XRD pattern of the obtained black powder shows the presence of the intence peak corresponding to (311) crystallographic orientation of the spinel phase of Fe3O4 magnetite nanoparticle. The mean size of the nanoparticles determined using the Scherrer formula, was in the order of 11nm, which is close to the value measured from Transmission Electron Microscopy which was found in order of 12nm. Magnetic properties of the obtained nanoparticles were determined from the hysteresis loop of magnetization vs the field, which shows a behavior of superparamagnetic nanoparticles which are a good candidate for nanomedicine.

Resume : Cancer is the main cause of mortality in the world. It is crucial to be able to design and, most importantly quantify an efficient therapeutic approach based on multimodal nanoparticles (NPs). Inorganic NPs based on metal oxides (e.g. TiO2 and Hf02) exhibiting several functionalities allowing their localization using various techniques (MRI, luminescence) as well as their quantification (IAB). When these metal oxides are bombarded with ionizing radiation, very large numbers of electrons are generated, considerably amplifying the lethal dose of energy in a tumor. The efficacy of the radiotherapy is thus multiplied, while the dose of irradiation impinging healthy tissues remains low. Two mechanisms may be involved, radioenhancement or radiosensitizer effect. In this context, NPs should be designed with several functionalities such as the ability to cross biological barriers as well as their non-toxicity for the humans. Moreover, it is important to be able to detect them in the human body, especially in the tumor target, via their magnetic or luminescence properties. For this purpose, rare-earth elements are inserted in the metal oxide matrices. As the lethal temperature for healthy and tumor cells differs, it is also important to ascertain the temperature of the cells during ionizing radiation treatment. This may be achieved based on the light emission of suitable pairs of rare-earth elements.

Resume : Background Calcium phosphate nanoparticles (CPNPs) can be used for applications in dental/medical treatments as they are biocompatible, biodegradable and their synthesis can be controlled. Here, we describe the preparation of CPNPs to be used for diffusion studies in hard tissues. We used near-infrared dye, Indocyanine Green (ICG) encapsulated within the nanostructure as a label for the detection of their diffusion through defective calcium phosphate substrates. Methods CPNPs were synthesised with co-precipitation of calcium chloride and sodium phosphate, in a solution of ICG for entrapment. Sodium citrate was then added as a capping agent. Their physicochemical properties, including particle size, zeta potential, morphology and the entrapment efficiency of ICG were characterised. Enamel slabs were then prepared and decalcified to simulate carious lesion. The ability of CPNPs to penetrate carious lesion was examined by staining the porous enamel with the nanoparticles. Results CPNPs were successfully made with an average particle size of 76 ± 14 nm using 300 mM citrate. They had an average zeta potential of – 21 ± 1.1 mV. The entrapment efficiency of ICG was 17%. Spherical and rice-shaped nanoparticles were observed under the AFM. Finally, selective accumulation of calcium phosphate nanoparticles on the decalcified areas of the teeth was observed using near-infrared imaging. At a longer contact period, more CPNPs had accumulated in the porous apatite based substrate, exhibited by a brighter fluorescence on the decalcified area. Conclusion CPNPs doped with ICG showed clear potential to detect and treat defects in hard tissues. For the next stage, we will find out the depth and time dependency of nanoparticle penetration into lesions.

Resume : Non-enzymatic glucose sensing is based on the direct catalytic oxidation of glucose and typically allows very high sensitivity at the expense of high fabrication cost, low selectivity and poor stability. Here we report on a novel NiO nanostructure fabricated by low-cost methods and successfully applied for non-enzymatic glucose sensing [1]. A Ni nanofoam (surface area of 25 m2/g) is obtained by room-temperature chemical bath deposition followed by 350 °C annealing in reducing ambient. To activate glucose detection a catalytic layer of Ni(OH)2/NiOOH is formed onto the nanofoam by cyclic voltammetry (CV). Two glucose sensor electrodes, on flexible and on rigid substrate, were fabricated, both showing sensitivity in the range of 1.5-3 mA/cm2mM, linear range of 0.01?0.7 mM, limit of detection (LOD) of 5 uM, resistance to chloride poisoning and excellent long-term stability (4% decrease in sensitivity after 64 days) and selectivity in the presence of common interfering species. Then, a full device was realized with a miniaturized electrochemical cell, based on a silicon chip with planar microelectrodes, connected to a PCB board (for the electrical driving and reading) and containing also a resistor for sensing test at high temperature. The sensing electrode was based on Ni oxy-hydroxide and glucose testing was performed with NaOH or PBS electrolytes. The device sensitivity decreases with decreasing the electrolyte pH, with still some activity also at the physiological pH of 7.4. [1] Iwu et al., Sensors and Actuators B 224 (2016) 764?771

Resume : GaN and related alloys such as AlGaN have attracted a great deal of attention due to their potential for wide-band gap optoelectronic and medical diagnostics applications. In the present work, AlGaN/GaN epitaxial films have been grown on SiN surface-treated sapphire substrate by metal organic vapor phase epitaxy (MOVPE) under atmospheric-pressure. The Al content in the studied samples is varied between 3% and 15% though the change of the flow rate of trimethylaluminum (TMA) from 12 µmol/min to 32 µmol/min. The optical properties and carrier dynamics of the AlGaN/GaN epitaxial films are investigated by means of time-resolved Photoluminescence (TR-PL). Low temperature measurements show that the PL emission peak broadens and shifts to higher energies when the TMA flow rate is increased. Simultaneously, the PL decay time increases with increasing Al contents in the studied films. These observations can be attributed to the carrier localization effects in band tail states due to compositional fluctuations in the AlGaN layers. Remarkably, the PL spectrum of the epitaxial film with a relatively high Al content of 15 % reveals several peaks. In order to find out about the origin of these features, temperature-dependent PL measurements are performed in the range from 25 K to 300 K. Keywords: AlGaN, Metal organic vapor phase epitaxy, Photoluminescence, time-resolved photoluminescence.

Resume : The Turkevich method is widely used to synthesize charge stabilised gold nanoparticles (Au NPs) from tetrachloroauric acid by reduction with sodium citrate. The low toxicity and ease at which such Au NPs can be funtionalised have made them very popular for biological applications. However this method is typically limited to producing larger Au NPs in excess of 10 nm. In order to achieve Au NPs which are 5 nm or less, other techniques are typically adopted which rely on the use of powerful surfactants or capping agents to limit particle growth. These particles in turn have more limited applications since the removal of the capping agent is not easily achieved. In our study, the effect of circular capillary microreactor on the size of Au NPs prepared by classic citrate synthesis method has been investigated. Since it is known that microreactor enhances both mass and heat transfer, a faster nucleation rate was expected, which would in turn lead to smaller particle sizes. By studying the effect of different parameters (temperature, initial pH and concentration of precursor and microreactor inner diameter), we have been able to produce Au NPs with mean sizes as low as 3 nm. There is still potential to improve on this result, as more parameters such as microreactor material, mixing method and residence time distribution will be studied in the future.

Resume : < p align="justify">The increase of multidrug-resistant bacteria infections represents an important bio-medical problem that requires the development of new antibacterial agents for which pathogens will not be able to develop resistance. The photodynamic (PDT) and/or photothermal (PTT) therapy can be considered as a viable alternative to conventional antibiotic therapies, which are becoming less efficient for the inactivation of microorganisms. PDT employs a photosensitizer (PS) molecule to induce irreversible cell damage upon laser irradiation at a specific wavelength. Near infrared (NIR) excitation light in the range of 700 to 1000 nm is of particular interest, because absorption of NIR photons in biological tissues is minimal and the penetration depth is optimal. In this study, we demonstrate that NIR plasmonic nanostructures, composed of silica coated gold nanorods with embedded indocyanine green (Au NRs@SiO< sub>2< /sub>-ICG) exhibit no cytotoxicity to cells and high capacity to inactivate a virulent strain of < i>Escherichia coli (E. coli)< /i> associated with Crohn?s disease under 810 nm pulsed-mode laser irradiation.< /p>

Resume : Ceria nanoparticles have been successfully synthesized using a new facile and eco-friendly sol-gel method. This approach is based on the thermal decomposition of ceric ammonium nitrate Ce(NH4)2(NO3)6 in the presence of a sucrose-pectin mixture. The chemical nature, structure and morphology of the CeO2 nanoparticles have been investigated by FT-IR, XPS, XRD and TEM. The XRD investigations reveal the formation of the fluorite crystal structure, while the TEM images show the formation of spherical nanoparticles with an average particle size range between 5 to 10 nm. Nanoceria switches between Ce4+ and Ce3+ states creating oxygen vacancies and this capability is similar to that of biological antioxidants. Thus, the antibacterial effect of the nanoparticles against four pathogenic bacteria: Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Bacillus cereus has been studied. More over, the cytotoxic effect induced by the ceria nanoparticles has been evaluated on two cell lines, D407 (human retinal pigment epithelial) and B16-F10 (melanoma murine). The results make the as-obtained ceria nanoparticles promising candidates for biomedical applications. Acknowledgments: This research was supported by UEFISCDI Project No. PN-II-RU-TE-2014-4-0944, Contract No 16/01.10.2015

Resume : We report on the fabrication of two types of Fe3O4 / para-aminobenzoic acid core-shell nanoparticles covered with a secondary intermediate silver layer. The final organic shells were built by phthalocyanine and flavonoid type compounds, respectively. High quality thin films of the final assembled core-shell nanomaterials have been obtained by MAPLE (matrix-assisted pulsed laser evaporation) and ESI (electrospray) deposition methods, on silicon and optical glass substrates. The intermediate and final nanostructures have been characterized by XRD, FT-IR, UV-VIS, HPLC-MS, DLS, HRTEM. MAPLE- and ESI-deposited thin coatings have been characterized by FT-IR microscopy, SEM, and AFM. The core-shell magnetic nanoparticles and obtained thin films have been investigated in vitro for their microbicidal (minimal inhibitory concentration assay) and anti-biofilm (microtiter assay of the minimal biofilm eradication concentration) properties on a large panel of bacterial and fungal strains antioxidant activity, as well as for their influence on the mammalian cells morphology, viability and proliferation (cytotoxicity, cellular cycle assay). Our results revealed that the multifunctional nanostructures proved both antioxidant and antimicrobial activities and a good biocompatibility. Therefore, the obtained thin films could be further used for the development of novel bioactive materials, with antimicrobial and antioxidant properties, that could be useful for a wide range of applications, in the prevention, control or eradication of the initial adherence and further formation of microbial biofilms, frequently involved in the microbial contamination of different media and products, as well as in the etiology of chronic and hard to treat infections.

Resume : In this contribution we demonstrated the possibility to obtain ultra-pure inorganic particles based-on Si-NPs by femtosecond laser assisted method in aqueous solution. Their physico-chemical properties were rigorously characterized by numerous technics such as TEM-XRD, XPS, and DLS/Zeta potential method. The results showed that Si-NPs appeared as crystalized Si spherical form with narrow size distribution around 50 nm and with surface charge close to -35 mV. In addition, by monitoring the dissolved oxygen in synthesis medium we demonstrated the possibility to control their stability duration in biological matrix by modulate their thickness oxide shell (SiOx) on their surface. Concerning their biological properties, in vitro tests were performed on human microvascular endothelial cells (HMEC) in order to assess their safety/cytotoxicity properties. Only 20 % decrease in cell survival was observed after 72h of incubation up to 50 µg/mL. In addition, their fate in term of biodistribution and biodegradability was completely assessed throw a systematic in vivo study realized on nude mice. The physiological and histological analysis showed that the Si-NPs not induce oxidative stress damage. These encouraging results contribute to develop promising biocompatible and biodegradable nanosystems for potential applications in nanomedicine.

Resume : The emergence of novel materials and processing at the nanoscale has set the conditions for the fabrication of a wide range of nano-objects and multilevel nanostructured networks. Here we report a simple and versatile waterborne synthesis of highly persistent superparamagnetic magnetic wires following the concept of electrostatic ??desalting transition?? [1]. The wires have diameters of 400 nm and lengths comprised between 1 and 500 µm. When submitted to a magnetic field, the wires behave as embedded rheometers and represent interesting sensors for micro-rheology. We develop the technique of rotational magnetic spectroscopy to measure the shear viscosity of simple and complex fluids, including water-gycerol mixtures, viscoelastic surfactant solutions and polysaccharide gels. With increasing frequency, the magnetic wires exhibit a rotational instability between a synchronous and an asynchronous regime of rotation. The critical frequency between these regimes varies as ?C ~ H^2/?L^2 where H is the magnetic excitation, ? the viscosity and L the wire length, and provides an accurate measure of the static shear viscosity. Fluids of biological relevance, e.g. intracellular medium of cells or pulmonary surfactant are also studied, showing that the technique can be useful for biomechanical studies [3]. [1] J. Fresnais, J.-F. Berret, B. Frka-Petesic, O. Sandre, R. Perzynski, Adv. Mater. 20 (2008) 3877 ? 3881 [2] L. Chevry, N. K. Sampathkumar, A. Cebers and J.-F. Berret, , Phys. Rev. E 88, 062306 (2013) [3] J.-F. Berret, Nature Comm. 7, 10134 (2016)

Resume : Colloidal stability in physiological fluids is often difficult to achieve, mainly due to their high salt concentration. Polymeric stabilisers are commonly used to tackle this problem, e.g. pegylation. More recently, macro-chain transfer agent (macro-CTA), which consists in a highly hydrophilic homopolymer synthesized via Reversible Addition-Fragmentation Transfer (RAFT) polymerization, have been used as steric stabilizer for the aqueous emulsion polymerization of hydrophobic co-monomers with > 90 % incorporation. This technique has been widely used to obtain self-stabilised particles (or ?hairy particles?), due to its well-defined architecture and nature, in a broad variety of ?high-tech? application areas such as optical data storage, security encryption, photonic crystals, and biotechnological or medical applications. The apparent core-shell structure of these hairy particles allows the periphery of the particles to be easily functionalised. Self-stabilized particles prepared by emulsion polymerization using poly[oligo(ethylene glycol) methacrylate] macro-CTA as polymeric stabilizer were successfully produced by varying reaction parameters such as co-monomer mixture, type of radical initiators, and amount of macro-CTA stabiliser. The resulting latex particles have been characterized in terms of particle size, Zeta potential, colloidal stability, Tg and amount of stabilisers. MTS and LIVE/DEAD assay have demonstrated the low toxicity of these self-stabilized particles.

Resume : Multi-modal imaging is an emerging area that integrates multiple imaging modalities to simultaneously capture visual information over many spatial scales. Luminescence imaging is a well-established, fast visualization technique, which can be easily adapted using portable devices and used for real-time surgical guidance. Compared to ultraviolet and visible light, short wavelength infrared (SWIR) has deeper penetration depths, which facilitates resolving anatomical features located up to 1 cm beneath the skin. In contrast, penetration depths of up to 5 cm can be achieved by using an emerging optical imaging modality known as photoacoustic (PA) imaging. PA imaging that converts laser energy into acoustic waves can be detected using ultrasound and used to image tissues with a high spatial resolution and better contrast at deeper penetration depth because ultrasound scattering is two to three orders of magnitude weaker than UV and visible optical scattering in tissues. Rare-earth doped CaF2 nanophosphors with enhanced SWIR emission have a promising potential for biomedical applications in diagnostics and deep tissue imaging due to their low vibrational energy, large optical transparency and superior biocompatibility. Highly emissive SWIR-emitting CaF2 nanophosphors is critical to enhancing the luminescence detection resolution and achieving deeper penetration depth for deep tissue imaging. The PA contrast that can be achieved using these CaF2 nanophosphors depends strongly on the absorption and vibrational characteristics as well as particle size and morphology. In our presentation, we will discuss the synthesis of CaF2 nanophosphors with enhanced SWIR emission using a tri-dopant scheme incorporating Ce, Yb and Er rare earth dopants. Next, the luminescence and PA properties of these as-synthesized CaF2 nanophosphors were measured to evaluate their feasibility as dual-modal contrast agents for luminescence and PA imaging to achieve high contrast, temporal and spatial resolution and deeper detection depth.

Resume : Therapeutic proteins are widely used in the field of diseases treatment due to their high specificity and bioactivity. A major disadvantage of proteins is their instability to external stressors during storage, transport and use. To overcome the problems, we report site-specific in situ growth of a polymer from a cyclized protein with improved in vitro stability and in vivo tumor retention. A linear green fluorescence protein (l-GFP) was genetically engineered, where two sortase A recognition substrates, pentaglycine and LPETG were fused to the N- and C-termini of GFP for sortase-catalyzed cyclization, a free cysteine residue was introduced near the C-terminus for site-specific installation of the atom transfer radical polymerization (ATRP) initiator. The maleimide-functionalized initiator was selectively attached to the cysteine residue to form a macroinitiator (c-GFP-Br), followed by in situ ATRP of oligo(ethylene glycol) methyl ether methacrylate (OEGMA) to generate a site-specific (C-terminal) and stoichiometric (1:1) c-GFP-POEGMA conjugate with almost quantitative conversion. Notably, the c-GFP-POEGMA conjugate displayed 9- and 310-fold increases in thermal stability compared with l-GFP and l-GFP-POEGMA, respectively, indicating that c-GFP-POEGMA was much more resistant to thermal denaturation than l-GFP and l-GFP-POEGMA. Besides, in a tumor-bearing mice model, the c-GFP-POEGMA conjugate showed significantly improved tumor retention than the l-GFP and l-GFP-POEGMA. The fluorescence of l-GFP in tumors rapidly decreased to nearly zero within 3 h, whereas the fluorescence of c-GFP-POEGMA and l-GFP-POEGMA in tumors slightly decreased and was detectable even after 48 h post administration. The fluorescence of c-GFP-POEGMA in tumors was over 6-fold higher than that of l-GFP-POEGMA at 60 h post injection, which might be attributed to the improved tumor retention of c-GFP over l-GFP. The data indicated that c-GFP-POEGMA had much better tumor retention than l-GFP and l-GFP-POEGMA. In conclusion, we develop a unique and general route to synthesize a site-specific and stoichiometric cyclic protein polymer conjugate with improved in vitro and in vivo stability via sortase-catalyzed cyclization followed by site-specific in situ ATRP.

Resume : Graphene and its derivatives (graphene oxides, nanoplatelets) have attracted research interest for their potential applications in nanomedicine, biotechnologies, pharmacology. Earlier studies focused on toxicity and biocompatibility. However, little information of their antioxidant activity and relation with different molecules well recognized as free radical scavengers such as OH. Herein are performed a comprehensive study on the antioxidant activity, measured by chemiluminescence, for a couple of hosts (graphene, graphene oxide) and guests (flavone-quercetin, rutin; flavanone-hesperidin, naringenin) selfassembled in high density ultrasonic field. The results have shown a high specific activity for graphene oxide and no effect for graphene. Thai is a consequence of the functional groups COOH and OH induced by solvo-thermal exfoliation during oxidation process of the carbon raw materials. In addition is proved that free radical scavenging is as mediated by proton transfer to the couple host-guest and not via electron transfer.

Resume : Preparation of well-defined nanoparticles is a big challenge for many scientists. Especially in case when structure is divided into the core and shell. However, this case is most interesting because of possibility to combine many properties in one low dimensional structure. Presented in this paper particles of core-shell structure have many interesting properties, among other we can stress: specific optical, magnetic, electrical, and mechanical one. In addition, few of them can be combined in one spices [1]. Discussed core-shell ferrite nanoparticles, has various composition of the core -mixture of Fe3+, Fe2+ and Men+ ions (where Me = Co2+, Co3+, Mn2+, Mn3+ or Ni2+). Elements were mixed in such way, that crystalline structure of magnetite is preserved. Shell layer was composed of magnetite. Applied synthesis was based on step-by-step seeds method with use of proper metal acetylacetonate salts in organic solution [2]. Characterization of the nanoparticles was done by Transmission Electron Microscopy, X-ray diffraction, Infrared spectroscopy, and Mössbauer spectroscopy.

Resume : In the present study, we have synthesized nanoparticles of iron ferrite by the sol-gel method with Polyethylene glycol (PEG-4000) as solvent and ferric iron nitrate Fe(NO3)3 as presursor with different concentrations. Above a temperature of 100°C, Fe3O4 was obtained directly without calcination. XRD pattern shows the presence of an intense peak corresponding to the known (311) crystallographic orientation of the Fe3O4 magnetite spinel phase. When followed by heat treatment at 400°C under atmospheric pressure, a phase transfomation to α- Fe2O3 was noticed due to the presence of the (104) crystallographic orientation in the XRD pattern. A mean size of about 12 nm of the magnetite nanoparticles was directly measured by TEM experiments, which is consistent with the value estimated from XRD analysis. Magnetic properties of the synthesized magnetite nanoparticles, measured using a SQUID magnetometer, showed a superpamagnetic behavior. All these features make the synthesized nanoparticles good candidates in nanomedicine.

Resume : Aim: to develop, characterize and evaluate an iron oxide nanosystem for the efficient stabilization and delivery of natural volatile compounds with antimicrobial effect. Materials and methods: Iron oxide nanoparticles were prepared by an adapted co-precipitation method and functionalized with limonene. The obtained nanosystem was characterized by High Resolution Transmission Electron Microscopy (HR-TEM), Scanning Electron Microscopy (SEM), Thermogravimetric analysis (TGA) and X-ray diffraction (XRD). The antimicrobial effect was tested for both free-floating (agar diffusion test and flow cytometry) and sessile bacterial cells belonging to Pseudomonas aeruginosa clinical and reference strains (Cravioto, microtiter and viable counts assay methods). Results: the obtained nanobiosystem inhibited the development of both free-floating and attached microbial cells in a strain and dose-dependent manner, as revealed by: i) the decrease of the minimum inhibitory concentration of the active compound by up to two folds, the most significant effect being observed for the antibiotic susceptible P. aeruginosa strains; ii) the reduced biofilm formation of all tested strains, but however a much higher amount of bioactive nanosystem being necessary to inhibit biofilm formation in case of antibiotic resistant strains; iii) modulation of the efflux pumps in a dose and strain dependent manner, limonene functionalized nanoparticles exhibiting a synergic effect with antibiotics.

Resume : Transportation systems based on albumin nanoparticles represent an important strategy since significant amounts of drug can be incorporated within the particle, as a function of different albumin binding sites. Bovine Serum Albumin (BSA) is highly water soluble and binds noncovalently with drugs or an inorganic material for the efficient drug delivery in various areas the affected body. Historically, albumin has been extensively applied as biodegradable anti-cancer drug charger because of its excellent biocompatibility and high stability in blood can accumulate in malignant or inflamed tissue. [1] [2] [3] Due to the well defined structure of the protein, containing charged amino acids, the albumin nanoparticles may allow electrostatic adsorption molecules negatively or positively charged without the application of other compounds on the surface. Therefore significant amounts of drug can be incorporated within the particle, as a function of different albumin binding sites. In this work we study the synthesis of bovine serum albumin nanoparticles, by desolvation process, containing salicylic acid as the active agent. Salicylic acid (SA) and salicylates are components of various plants and have been used for medication with anti-inflammatory, antibacterial and antifungal properties. Commercially, salicylic acid salicylates are an active metabolite of aspirin (acetyl salicylic acid) and have been used since century V. However, when administered orally to adults (usual dose provided by the manufacturer), decomposes 50% salicylates. [4] Then, over time there is a search to develop new systems in order to improve the bioavailability of SA and salicylates in the human body. To this end, in synthetic procedure, the pH ranged synthesis (5.4, 7.4 and 9.0) in order to evaluate the influence on the size and stability of the formed nanoparticles. The samples were analyzed using by FE-SEM, TEM, FTIR, Zeta Potential and DLS. Through fluorescence was possible to analyze the release of salicylic acid "in vitro" in PBS solution. The chemical morphology characterization results and the “in vitro” release studies indicate the potential use of these nanoparticles, as drug carriers in biological systems requiring a fast release of salicylic acid. Keywords: albumin nanoparticles, salicylic acid entrapped, drug delivery; [1] R. Xu, M. Fisher, R.L. Juliano, Bioconjug. Chem. 22 (2011) 870. [2] G.V. Patil, Drug Dev. Res. 58 (2003) 219. [3] J.M. Irache, M. Merodio, A. Arnedo, M.A. Camapanero, M. Mirshahi, S. Espuelas, Mini Rev. Med. Chem. 5 (2005) 293. [4] M. A. Rogers, Y. F. Yan, K. Ben-Elazar, Y. Lan, J. Faig, K. Smith, K. E. Uhrich, Biomacromolecules, 2014, 15, 3406−3411. eriuhle@fc.unesp.br – Av. Eng. Luiz Edmundo Carrijo Coube, 14-01, 17033-360 Bauru – SP – Brazil Tel: +55-14-3103-6084, ramal. 34; Fax: +55-14-31036094.

Resume : Iron oxide nanoparticles, NP, mineralized within human H-chain ferritin, HFt, represent a viable platform to develop novel theranostic agents, as they offer multiple advantages in terms of stability, biocompatibility and easiness of functionalization. However, a major limitation is that the NP size cannot exceed the inner diameter (ca. 8 nm), a too low size to produce a sizable amount of heat. In order to overcome this drawback while still exploiting the advantages provided by HFt, we developed two different approaches: first, we increased the magnetic anisotropy of the NPs by doping with Co2+ ions (CoNP-HFt); second, we conjugated HFt on larger (15 nm), chemically synthesized, maghemite NPs (HFt-NP). In vitro tests showed that both constructs were highly biocompatible. The hyperthermic properties of CoNP-HFt and HFt-NPs were investigated through calorimetric technique and related to structural and magnetic properties. A doping of 5% was found to enhance the hyperthermic efficiency of CoNP-HFt and to induce a significant reduction of cell viability when exposed to an alternating field. On the other hand, despite of the much larger hyperthermic efficiency, HFt-NP were found to affect cell viability only when internalized in large amount, as confirmed by confocal microscopy. These results enlighten the fundamental role of cell-NP interactions with respect to large hyperthermic efficiency. Research funded by RINAME (MIUR), BaTMAN (CARIPLO n.2013-0752) and CNR-CNPq projects.

Resume : We report the assemblies of novel nanohybrid materials formed between planar carbon systems (PCS) such as coronene or TRGO and a series of new halogenated ?- amino acid tagged NDI chromophores. The functionalization of graphene-like systems with NDI chromophores can be facilitated via a mechanism of Föster resonance energy transfer (FRET) whereby photoinduced electron transfer takes place. UV-Vis, Fluorescence spectroscopies and fluorescence lifetime decay measurements rationalised such FRET mechanisms and elucidated the role of coronene and TRGO as a quenching materials. The morphology of organic nanoparticles generated from the complexation of NDIs with coronene and TRGO nanosheets were also studied by SEM and TEM. NMR experiments allowed for studies into the kinetics of the supramolecular association between chromophores and PCS. Confocal fluorescence microscopy studies carried out on NDI-coronene and NDI-TRGO show that our nanocomposites are up-taken in PC-3 cells suggesting their potential as prostate cancer bioimaging agents. MTT assays suggest that the deliberate functionalization of the surface of TRGO and coronene with phenylalanine NDIs significantly improves the biocompatibility of such graphene-like nanocomposites with respect to the pristine TRGO and coronene. The results here presented may lead to a new prospective in the use of the functionalization processes to achieve fluorogenic composite materials based on planer carbon materials for biological application The authors would like to thanks ERC Consolidator grant scheme (O2Sense to Sofia Pascu).

Resume : In this presentation I will the most recent results of our group on synthesis and functionalisation of nanoparticles for biomedical applications. Ref: 1. R. Hachani, M. Lowdell, M. Birchall, A. Hervault, D. Merts, S. Begin-Colin, N.T.K. Thanh*. (2016) Polyol synthesis, functionalisation, and biocompatibility studies of superparamagnetic iron oxide nanoparticles for potential MRI contrast agents. Nanoscale. DOI: 10.1039/c5nr03867g. Open Access 2. R. M. Pallares, X. Su, S. H. Lim, N. T. K Thanh* (2016) Fine-Tuning Gold Nanorods Dimensions and Plasmonic Properties Using the Hofmeister Salt Effects. Journal of Material Chemistry C. 4: 53-61. Open access. Front Cover 3. C. Blanco-Andujar, P. Southern, D. Ortega, S.A. Nesbitt, Q.A., Pankhurst and Thanh, N. T. K*. (2016) Real -time tracking of delayed-onset cellular apoptosis induce d by intracellular magnetic hyperthermia. Nanomedicine. 11: 121-136. Open Access. 4. R. M. Pallares, S. L. Kong, H. R. Tan, Thanh, N.T.K, Y. Lu and X. Su (2015) A plasmonic nanosensor with inverse sensitivity for circulating cell-free DNA quantification. Chemical Communications. 51, 14524 - 14527 5. L. T. Lu, N. T. Dung, L. D. Tung, C. T. Thanh, O. K Quy, N. V. Chuc and N. T. K. Thanh* (2015) Synthesis of magnetic cobalt ferrite nanoparticles with controlled morphology, monodispersity and composition: the influence of solvent, surfactant, reductant and synthetic condition. Nanoscale. 7: 19596-19610. Open Access. Front Cover 6. R. Baber, L. Mazzei, N. T. K. Thanh, A. Gavriilidis (2015) Synthesis of silver nanoparticles in microfluidic coaxial flow reactors. RSC Advances. 5: 95585-95591. Open Acess

Resume : Multifunctional thermoresponsive nanoparticles were easily designed via activators regenerated by electron transfer atom transfer radical polymerization (ARGET- ATRP) of commercially available methacrylate derivatives from the surface of zinc oxide quantum dots (ZnO QDs). Conserving the luminescent properties of ZnO QDs, these nanoparticles showed different responsive behavior in water as well as in different media by varying the composition of the copolymer. The results also showed that below a specific concentration these nanoparticles have a very low toxicity towards healthy cells. Therefore, they could be very interesting candidates for applications in nanomedicine.

Resume : Gynaecological cancers include ovarian cancer and endometrial cancer. We have developed biological and synthetic nanomedicines to target these two diseases. To effectively diagnose and monitor cancer effective nano-based sensors are required. We have developed a graphene based sensor that offers levels of sensitivity enabling detection of circulating cancer markers including epigenetic factors. The development and integration of nanomedicines and nanodiagnostics will be discussed, including their fit with the emerging global personalised medicine agenda.

Resume : The fabrication of nanoporous / nanoparticulate composites and their applications via surface patterning with chemicals and bio-chemicals has a direct impact in bio-sensing and bio-separation. Surface patterning on nanoparticles in suspension can be a complex process due to the aggregation of the particles and their Brownian motion in the suspension. An overview of group?s research on nanomaterials and their applications in the separation of nucleic acids (DNA and RNA) from the biological cells will be presented in connection with an industrial collaboration with Q-Bioanalytic, Germany. The possibility of affinity interaction of biomolecules i.e. nucleic acid, protein, antibody, microorganisms etc. through hybrid capture will also be discussed in the context of food quality and hygiene in Bio-sensing which has recently been published in Nature publishing group (http://www.nature.com/srep/2012/120807/srep00564/full/srep00564.html?WT.ec_id=SREP-639-20120903). Separation of toxic and microbial contaminants from water and soil using nanotechnology tool will be discussed in the context of on-going multinational projects (http://senlabs.org/international-projects/ & http://nanowateratuclan.org/) in collaboration with top academic and industrial researchers from Europe, India and China. Recent development (UK Patent: 2013: GB1315407.5. & PCT/GB2014/052,630) on sensing antimicrobial nanocomposites will be discussed in connection with water technology.

Resume : We have developed a unique DNA-assembled gold nanoparticles (AuNPs) dimer for biosensing via the dynamic light scattering (DLS) platform. Using estrogen receptor (ER), which binds specifically to a double-stranded (ds) DNA sequence containing estrogen response element (ERE), as an example, we successfully incorporated ERE sequence into the DNA linkers to bridge the AuNPs dimer for ER binding. Coupled with DLS, this AuNP dimer-based DLS detection system gave distinct readout of a single ?complex peak? in the presence of the target molecule (i.e., ER). This unique signature marked the first time that such nanostructures can be used to study transcription factor-DNA interactions, which DLS alone cannot do. This was unlike previously reported AuNP-DLS assays that gave random and broad distribution of particles size upon target binding. In addition, the ERE-containing AuNP dimers could also suppress the light-scattering signal from the unbound proteins and other interfering factors (e.g., buffer background), and has potential for sensitive detection of target proteins in complex biological samples such as cell lysates. In short, the as-developed AuNP dimer probe coupled with DLS is a simple (mix and test), rapid (readout in ~5min) and sensitive (low nM levels of ER) platform to detect sequence-specific protein-DNA binding event.

Resume : To explore the most dynamic cross field of nanotechnology and life science, and to find a more practical method of fabricating nanodevices on a very large scale with advantages of low manufacturing cost, high reliability and reproducibility, we successfully fabricated a submicron IrO2 nanowire array biosensor platform by conventional complementary metal-oxide-semiconductor (CMOS) process. Single crystal IrO2 nanowire array was grown uniformly on a 6-in. wafer surface by chemical vapor deposition (CVD) method and patterned into submicron array clusters. The obtained clusters were positioned in a designed pattern similar to a multiple electrode array (MEA) format, and each was individually addressed. The fabrication method was found to be reliable, low cost and robust. The final chip showed excellent transparency, functionality and durability. These chips have been used for neuron cell stimulation and protein sensing, the experiment results will be presented.

Resume : SPIONs have exciting potential as cancer theranostic agents. SPION can be used for imaging and also induced to heat by application of an alternating magnetic field. Heat is toxic to the targeted cells, resulting in effective localized hyperthermic therapy. The feasibility and translational challenges of exploiting this nanomedicine approach to cancer treatment will be discussed.

Resume : The clinical exploitation of plasmonic particles such as gold nanorods, nanoshells and nanocages has become a remarkable mission for a broad community of scientists. These particles are proposed as contrast agents for various applications in biomedical optics, including the photothermal ablation and the photoacoustic imaging of cancer, which are emerging as hopeful alternatives in oncology. At present, one of the most relevant challenges in nanomedicine is to really optimize the delivery of functional nanoparticles into tumors upon systemic administration, as a ?magic bullet?. In this context, PEGylated gold nanorods offer favorable conditions. Their hydrodynamic sizes are ideal to exploit the enhanced permeability and retention of most tumors. Besides, PEG strands enable their bio-conjugation with targeting units, such as antibodies. PEGylated and bio-conjugated gold nanorods display high specificity and blood compatibility in vitro. Therefore, their direct injection into the bloodstream has become a mainstream solution. However, after more than one decade of preclinical trials in dozens of labs, this route still remains problematic. Upon intravenous injection, more than 90% of the gold nanorods typically get captured by the mononuclear phagocyte system. More recently, the notion to exploit the natural tropism of cells such as tumor-associated macrophages, T cells, mesenchymal stem cells and neural stem cells has begun to emerge as a radical alternative. Tumor-tropic cells hold an innate ability to infiltrate tumors across all barriers with the aim to modulate their microenvironment and may be exploited as biological vehicles to take up and to deliver functional cargos. Several authors have suggested that this approach may improve the distribution of different particles both between the tumor and other parts of the body and locally within the tumor. In addition, the use of cellular vehicles implies a significant simplification of the biological interface of the plasmonic particles. Here, we disclose our recent achievements in the preparation of polycationic gold nanorods for a robust design of cellular vehicles that combine high optical absorbance, viability and chemotaxis. We discuss perspectives and possible limitations for their use as contrast agents for photoacoustic imaging, CT and photothermal treatments. We are confident that our work may inspire new solutions at the crossroads of nanomedicine and biomedical optics.

Resume : Nanoparticles are increasingly popular choices for labeling and tracking cells in biomedical applications such as cell therapy. However, all current types of nanoparticles fail to provide a real-time and noninvasive monitoring of cell status and functions while often generating false positive signals. Herein, a nanosensor platform to track the real-time expression of specific biomarkers that correlate with cell status and functions is reported. Nanosensors are synthesized by encapsulating selected sensor molecules within biodegradable polymeric nanoparticles. Upon intracellular entry, nanosensors reside within the cell cytoplasm, serving as a depot to continuously release sensor molecules for approximately 30 days. In the absence of the target biomarkers, the released sensor molecules remain ?Off?. When the biomarker(s) is expressed, a detectable signal is generated (On). As a proof-of-concepts, three nanosensor formulations based on this concept are synthesized to monitor cell viability, secretion of nitric oxide, and stem cell differentiation.

Resume : Bimetallic nanostructures show exciting potential as materials for effective photothermal hyperthermia therapy. This talk will discuss the seed-mediated synthesis of palladium-gold (Pd-Au) nanostructures containing multiple gold nanocrystals on highly-branched palladium seeds.[1] The nanostructures were synthesized via the addition of gold precursor to a palladium seed solution, in the presence of oleylamine which acts as both a reducing and stabilizing agent. The interaction and the electronic coupling between gold nanocrystals and between palladium and gold broadened and red-shifted the localized surface plasmon resonance (LSPR) absorption maximum of the gold nanocrystals into the near-infrared (NIR) region, to give enhanced suitability for photothermal hyperthermia therapy. Results discussed will show Pd-Au heterostructures irradiated with 808 nm laser light caused destruction of HeLa cancer cells in vitro, as well as complete destruction of tumor xenographs in mouse models in vivo for effective photothermal hyperthermia. [1] A. J. McGrath, Y-H. Chien, S. Cheong, D. A. J. Herman, J. Watt, A. M. Henning, L. O. Gloag, C-S. Yeh, R. D. Tilley, ‘Gold over Branched Palladium Nanostructures for Photothermal Cancer Therapy’ ACS Nano, 9,12283–1229, (2015).

Resume : In order to overcome the current limitations of systemic drug administrations in some clinical indications, Targeted Delivery Systems (TDS) emerge as a promising facet of the biomedical and pharmaceutical research and industry, since TDS allow controlling the location of the bioactive systems and producing efficient pharmacological effects only to specifically intended target sites, with significant therapeutic benefits and reduced side effects. In particular, the local drug release mediated by injectable TDS directly implanted on the target site, using interventional radiology procedures, is a simpler and short-term strategy, from a technical and regulatory point of view. Interestingly, it also anticipates the foreseeable increase in the use of interventional imaging machines in operating rooms. In this context, we will describe a new class of TDS for bone therapies, based on injectable calcium phosphate cements (CPCs) combined to drugs, with unique multitasking capabilities. In addition to its role as carrier to allow the controlled local elution of the drug, CPCs also act as a “calcium and phosphate ions reservoir” that promote bone repair in the bone site where it is injected. The expected feature is a high benefit/risk ratio, since only one minimally invasive injection should be necessary. Some examples will be given for the following indications: prevention of osteoporotic hip fracture, bone tumor therapy, spinal fusion.

Resume : Multimodal contrast agents (MCAs) exploit the benefit of multiple and complementary imaging techniques. Magnetic resonance imaging uses safe radiation and allows total body screening, but suffers on low spatial resolution if compared to optical imaging, that is characterized, however, by low tissue penetration. Surface Enhanced Raman Spectroscopy (SERS) is a vibrational technique amplified by a plasmonic substrates, namely noble metal nanoparticles. SERS allows imaging at sub-micrometer resolution with intensities comparable to fluorescence. Into this context we designed the synthesis of SuperDOTA, a new polyethylene glycol based polymer characterized by high Gd3+ loadings. SuperDOTA consists by three main fractions. The first is a cysteine amino acid, containing a thiol group for a stable linking to AuNPs surfaces. The second is a repetition of a short PEG spacer and a DOTA based branch for the Gd3+ chelation. The third is a long-chain PEG, to enhance the biocompatibility. Gold nanoparticles were synthesized by laser ablation synthesis in solvent (LASiS) and a SERS substrate is obtained by a controlled centrifugation. Si-Naphthalocyanine, used as SERS label, was functionalized with mercaptoethanol on both the apical sites to improve the optimal situation in which the dye molecule stays between two plasmonic particles, i.e. in a hot spot. The new nanostructure is used for multicontrast imaging using both MRI and SERS also for in-vivo measurements.

Resume : A reliable preclinical diagnosis of neurodegenerative diseases, such as Alzheimer’s disease (AD), is of great importance both for drug development and efficient treatment. Immunoassays are widely used for multiple sample analysis and quantification of biomarkers in biological samples. A promising biomarker for AD is the Amyloid beta (Abeta) oligomer, an aggregated but soluble multimeric form of the Abeta peptide which is the major culprit in AD pathology. Such oligomers can be detected by the sFIDA (surface-based fluorescence intensity distribution analysis) assay. A unique feature of the sFIDA assay is its ability not only to count single oligomers but also to analyze the composition of each particle. Therefore also hetero-aggregates consisting of more than one disease-associated peptide can be quantified. In this work we present a set of standard materials that can be used in oligomer-specific immunoassays. In contrast to other materials developed for oligomer-specific diagnostics the presented materials are tunable in their number of epitopes and epitope composition. A tunable epitope quantity and composition allows the design of hetero-aggregate mimicking particles in the oligomer size range. Therefore, immunoassay development and standardization can be performed towards the detection and quantification of hetero-aggregates within biological samples. We present the synthesis strategy of the nanoparticles and show the performance of these materials in the nanoparticle sensitive sFIDA assay on the example of hetero-aggregates consisting of Abeta and Tau protein.

Resume : Cancer is a leading cause of death worldwide [1]. Future projections predict globally an increase up to 23.6 million new cases each year by 2030. Therefore, cancer research cannot wait, and new approaches are urgently required. RADIOMAG aims at building up the next generation of researchers in a highly multidisciplinary field by enabling current and organising new research dealing with the efficiency of radiotherapy and its synergetic combination with magnetic fluid hyperthermia (MFH). The latter treatment takes advantage of the heat produced by magnetic particles in alternating magnetic fields to kill or damage cancer cells [2]. The last two decades have been quite prolific in the development of bespoke magnetic nanoparticles and instrumentation for MFH [3], but despite the recent positive outcomes from clinical trials with glioblastoma multiforme patients, the treatment is still far from becoming a standard practice. The presentation will give an overview about the activities of this COST Action and present early scientific results. Special emphasis will be put on the design of a large scale inter-laboratory experiment, the first of its kind in the pursuit of a more accurate and comparable characterisation of nanoparticles for magnetic fluid hyperthermia. 1. S.W. Bernard, WHO World Cancer Report, ISBN 978-92-832-0429-9, (2014) 2. A. Hervault et al, Nanoscale 6 (2014) 11553; S. Dutz, R. Hergtet al, Nanotechnology 25 (2014) 452001 3. E. Périgo et al, Appl. Phys. Rev. 2 (2015) 041302

Resume : Elastin-like proteins (ELP) are recombinant proteins of the VPGXG pentapeptide (X being any amino acid but proline), the sequence of elastin, an extracellular matrix protein that undergoes deswelling transition at a critical temperature. This communication reports the grafting of diblock ELPs designed with a thermosensitive block and a hydrophilic block proving steric repulsion, onto superparamagnetic iron oxide NPs. Magnetic cores of superior magnetic heating efficiency were prepared by the DEG-NMDEA route introduced by Caruntu et al. The functionalization of the magnetic NPs by mixed brushes of ELP and PEO chains resulted into stable magnetic and thermosensitive colloids with a decreasing temperature-size response. The ELP and PEO grafting was achieved by a convergent strategy using a phosphonate moiety introduced by a crosslinking agent grafted onto a cysteine residue near the N-terminus of the diblock ELP construction or on PEG-SH. In situ DLS measurements during magnetic hyperthermia (MH) indicated that diblock ELP-grafted iron oxide NPs exhibited fast size changes versus the applied field strength profile. At long times (thermal equilibrium), the brush thickness correlates with the macroscopic temperature measured by fiber optic thermometry. During the MH treatment, a transient gap between the macroscopic and the local temperature was also evidenced. These results could help understanding the ?cold hyperthermia? effect of iron oxide NPs in intra-cellular compartments.

Resume : Theranostic systems are developed to monitor a treatment in vivo according to their abilities for both imaging and therapy. We propose an innovative strategy to treat colon cancer with the design of magnetic thermosensitive liposomes that enable to target a superficial tumor under a magnetic field gradient and simultaneously to deliver an antitumoral drug by High Intensity Focused Ultrasounds (HIFU) application, with additional bimodal imaging properties for in vivo monitoring. In our study, we demonstrate the significant increase of accumulation of Ultra-Magnetic Liposomes (UML) in CT26 murine tumor using magnetic guidance compared with passive enhanced permeability retention effect. UMLs are highly loaded with magnetic nanoparticles of iron oxide (γ-Fe2O3) enabling both magnetic guidance and in vivo monitoring by MRI. The addition of a fluorescent probe in the formulation allows to track them by optical imaging. In vivo UML biodistribution was evaluated with dynamic susceptibility contrast imaging in MRI and fluorescent imaging. The accumulation of UMLs into the tumor after I.V. injection was measured using MRI image percentile processing, iron dosage by ICP AES and histology. Co-encapsulation of magnetic nanoparticles and SN38, an antitumoral drug, gave theranostic properties to our thermosensitive liposomes. Application of HIFU can, then, be used in the tumor for an active release of the drug in the region of interest for the development of an innovative therapy.

Resume : Key words: Magnetic nanoparticle; Liposome; drug combination; Magnetic nanoparticles have been widely investigated as a promising nano-medicine agent since last decade. A number of in vitro and in vivo results suggest the chemo-thermotherapy via multifunctional magnetic nanoparticle has positive effect in cancer treatment [1]. However, unlike the thermo-radiotherapy, the clinical trials of chemo-thermotherapy could not make a consistent synergism conclusion. [2] Apart from the complicated tumor architecture, it may due to the tumour heterogeneity. Herein, our group constructs a multifunctional magnetic thermo sensitive liposome and optimizes it by introducing both pharmaceutics and biomolecular concepts. In the first part, we refined the polyol method for synthesis the multi core magnetic nano-flower that has high magnetic property. In order to boost the chemo-thermo effect, the most synergistic chemodrugs combination ratio was carefully chosen among the 3 most prevalent anti breast cancer medicines: gemcitabine, paclitaxel and doxorubicin, by testing on different breast cancer cell lines. Both of them were loaded into the thermosensitve liposomes via thin film hydration methods. The efficiency of the multifunctional magnetic chemosensitive liposome will be verified and compared with the baseline among all cell lines. Based on the outcomes, the second part will be focusing on further optimizing our chemo-thermotherapy strategy via biomolecular analysis. Ref: [1] A. Hervault and N. T. K. Thanh, ?Magnetic nanoparticle-based therapeutic agents for thermo-chemotherapy treatment of cancer,? Nanoscale, vol. 6, no. 20, pp. 11553?11573, 2014. [2] B. Hildebrandt, P. Wust, O. Ahlers, A. Dieing, G. Sreenivasa, T. Kerner, R. Felix, and H. Riess, ?The cellular and molecular basis of hyperthermia,? vol. 43, pp. 33?56, 2002

Resume : Oligonucleotides (ODNs) present a high therapeutic potential for a wide variety of diseases. However, their clinical development is limited by their degradation by nucleases and their poor blood circulation time. Depending on the administration mode and the cellular target, these macromolecules will have to cross the vascular endothelium, to diffuse through the extracellular matrix, to be transport through the cell membrane and finally to reach the cytoplasm. To overcome these barriers, many strategies have been developed. Here, we report a method allowing the direct and one-step oligonucleotide conjugation onto iron oxide nanoparticle surface using complexation affinity of phosphate1,2 or phosphorothioate3 functions and avidity effect induced by the large number of these moieties on the ODN backbone for the nanoparticle surface. We show that the average number of ODN per nanoparticle could be controlled and that the adopted conformation was related to the grafting density progressing from a flat conformation to perpendicular one with the increase of ODN loading The phosphodiester ODN delivery in cells and the biological activity (inhibition of the nuclear localization of the transcription factor STAT3) seems to be related to the grafting density. Finally, we explore different strategies, based on the electrostatic interactions between the different partners, to functionalize the surface of SPIONs with a ODN and a cationic cell penetrating peptide3. References 1- Easily Controlled Grafting of Oligonucleotides on Fe2O3 Nanoparticles: Physico-Chemical Characterization of DNA Organization and Biological Activity Studies. F. Geinguenaud, I. Souissi, R.Fagard, Y. Lalatonne, L. Motte, J. Phys Chem. B 2014,1535-1544. 2- Electrostatic Assembly of a DNA Superparamagnetic Nanotool for Simultaneous Intracellular Delivery and In-Situ Monitoring, F. Geinguenaud, I. Souissi, R. Fagard, L. Motte, Y. Lalatonne, Nanomed. Nanotechnol., 8 (7), 2012,1106-15. 3- Iron oxide nanoparticles coated with a phosphorothioate oligonucleotide and a cationic peptide: exploring four different ways of surface functionalization. F. Geinguenaud, C. Banissi, A. Carpentier, L. Motte, Nanomaterials, Special Issue "Nanoparticles in Theranostics": invited article,5(4), 2015,1588-1609.

Resume : Biomedical applications of superparamagnetic iron oxide nanoparticles (SPIONs) require an optimized coating via which SPIONs interact with any biological entities (e.g., proteins, cell membranes). We demonstrate a spontaneous one-step way to form carboxylated/PEGylated shell on SPIONs by post-coating method, in which optimized magnetic cores were combined with commercial (e.g. poly(acrylic acid) and designed polymer molecules. In the latter, poly(ethylene glycol) methyl ether methacrylate (PEGMA) and acrylic acid (AA) were chemically coupled in a comb-like copolymer, P(PEGMA-co-AA) with PEG chains and pendant carboxylates in a single coating molecule. The multiple bindings on SPIONs via surface complexation of Fe-OH sites by carboxylates are spontaneous. The formed polyelectrolyte layer hinders SPIONs’ dissolution, provides combined electrostatic and steric (electrosteric) stabilization, and high protein repellency by PEG moieties. In addition, free carboxylates still exist in the shell for anchoring bioactive molecules via peptide bonding. In the lecture, we will show the synthesis and post-coating of naked SPIONs, their physicochemical and colloidal characterization and the comparison of optimized products’ testing for hemocompatibility (blood sedimentation rate, blood smear and blood cell viability), in vitro cell tests (MTT cell proliferation assay, Prussian blue staining), MRI contrast and hyperthermia efficiency.

Resume : For tissue repair of regeneration based on tissue engineering principle, scientists with medical and biological background often pay much attention to stem cell itself, or use some traditional biomaterials as scaffold, and other researchers from materials engineering filed often pay much attention to scaffold research and the last step, test repairing effect in an animal model by implant the scaffold. The most important part, interaction between the surface of biomaterials and differentiation of stem cells is often ignored, because it need interdiscipline background, and people rarely possesses very strong background in both biology and materials science. Fortunately, most recently, an increasing number of researchers devote to this field, and pay much attention to the relationship between materials and stem cells, which will make great progress in this field. The effect of nanotopography of the biomaterials on differentiation of stem cells is the first topic to study the interaction between materials and stem cells. However, most researches were taken the materials such as silicon, PDMS as basic materials, because they are very easy for building nanostructures by micro-manufacture techniques. Although these works uncovered some unknown phenomenon about the response of stem cells on nanotopography, they are very far for application in tissue engineering because of the composition difference among the experimental samples and biomaterials. Nanoparticles, a very important category of biomaterials, have been widely investigated and even applied in cell and tissue imaging, drug delivery, photothermal cancer therapy, etc. Normally, the nanoparticles should be injected into the body for their therapy function. After fulfilling their bio-function, the nanoparticles, especially the particles without bio-degradation ability will stay in the body. Some of the nanoparticles can be discharged thorough circulatory system, and others may enter the area of the stem cells. As is well known, stem cells has strong Recent years, the interaction between material and stem cell has been attract much attention, because it is helpful to understand some bio-process and very useful for design and preparation of tissue engineering scaffold for tissue repair and tissue regeneration. Nanotopography is one of the most important physical cues for regulation fate of stem cells. Because the extracellular matrix is composed by polysaccharide and protein fibers with nanoscale diameter, the nanotopography can be act as a physical signal to tune differentiation of stem cells by endocytosis ability, which can take a large number of nanoparticles. Although there are thousands researchers do research on the interaction between cancer cell and nanoparticles, a few scientists have involved in the effect of nanoparticle on differentiation of stem cells. However, the behavior of nanoparticles in stem cell is very important, which could affect the differentiation ability of the stem cell, and will further affect the maintain and reconstruction of some important tissues in the body. Therefore, the regulation effect of nanoparticles on differentiation of stem cells is a very important topic for biomaterials. With last three years, our group focuses on the interaction of nanostructures and nanoparticles and stem cells. We found some nanoarray and nanostructures built by biomaterials can enhance osteogenic and neural differentiation, and demonstrates that the localized and committed differentiation could be realized in one system by integrating materials with different topography. Moreover, we have found that some nanoparticles, such as graphene quantum dots, lithium niobate nanoparticle, not only have good cytocompatibility, but also can enhance osteogenic differentiation of MSCs, which proved the safety of some nanoparticles during the therapy applications. In this talk, we will report the above works, and try to explain the reasons for physical signal induced differentiation from both physical mechanism and bio pathways. We believe that the regulation effect of nanostructures and nanoparticle will have great impact for design and application of biomaterials, especially for tissue engineering scaffold, and will bring great progress in tissue regeneration medicine.

Resume : Nowadays, nanoparticles can be obtained by many various methods in variable forms. At the nanometric level properties of nanostructures are very sensitive to many parameters: composition, size, crystallinity, shape. In this presentation we would like to compare nanoparticles which were prepared by three methods. Chosen samples were fabricated by: i) co-percipitation of magnetite from iron chlorides in aqueous solution, ii) thermal decomposition of iron (III) acetylacetonate comples in organic solution; iii) laser ablation in solution. Each of the presented methods leads to other kind of ferrofluids and therefore its application is different. Laser Ablation Synthesis in Solution, involves few phenomena: firstly material is evaporated from bulk target, with use of focused lasers short impulses, melt reactions, and particles fabrication. By this method, bimetallic, but also metal oxide nanoparticles can be obtained. Chemical synthesis of nanoparticles in organic or inorganic solvent and proper stabilizer allows to ferrofluid fabrication. Every type of obtained nanoparticles will be characterized by TEM, IR, XRD, UV-Vis and Mössbauer spectroscopy.

Resume : The hybrid organic/inorganic nature of the surface of functionalized magnetic iron nanoparticles (MNPs) plays a pivotal role in determining their properties and in turn their applicability. In this contribution, the role of Fe3O4 - surface functionalization on MNPs capability of overcoming “in vitro” models of the blood-brain barrier (BBB) has been investigated. Three different coatings have been covalently anchored on the Fe3O4 surface through a multistep route based on MNPs prefunctionalization with a bifunctional phosphonic linker, (3-aminopropyl) phosphonic acid, able to bind on iron surface through the phosphonic groups. Then the functional molecules have been covalently bonded to the amino group of the phosphonic monolayer. In particular, attention has been focused on three functional systems: a simple polyethylene glycol (PEG) coating, a mixed folic acid – PEG layer and a coating based on gH625 a viral fusion peptide able to easily cross through the cell membrane. To monitor MNPs uptake into BBB, all the nanosystems have been marked with rhodamine- or NBD- based luminescent probes covalently bonded, respectively, to the phosphonic acid pre-functionalized nanoparticles or to the anchored gH625 peptide. Finally, using primary microvascular endothelial cells from human brain (EC), the effect of MNPs permeation on the production of reactive oxygen species (ROS) has been evaluated and compared for all the investigated nanosystems.