Bioinspired and biointegrated materials as new frontiers nanomaterials VIII

Resume : The interaction of single-walled carbon nanotubes (SWNTs) with DNA and of proteins with DNA are crucial for several nano-materials applications, including SWNT:DNA nanoscale devices or nanosized building blocks for use in nano-switches, nanoscale wiring and biomedical applications involving DNa delivery. In biotechnological applications, modulating the DNA:SWNT interaction is important for SWNT purification, DNA recognition, and ultrafast DNA sequencing, and drug delivery. I will present a study of the the conformational equilibrium and the dynamics between B-to-A forms of double-stranded DNA adsorbed onto single-walled carbon nanotubes (SWNT) using free energy profile calculations based on all-atom molecular dynamics simulations. The potential of mean force of the B-to-A transition of ds-DNA in the presence of an uncharged (10,0) carbon nanotube for two dodecamers with poly-AT or poly-GC sequences is calculated as a function of a root-mean-square-distance metric quantifying the B-to-A transition. The calculations reveal that in the presence of a SWNT DNA favors B-form DNA significantly in both poly-GC and poly-AT sequences. Furthermore, the poly-AT DNA:SWNT complex shows a higher energy penalty for adopting an A-like conformation than poly-GC DNA:SWNT by several kcal/mol. The presence of a SWNT on either poly-AT or poly-GC DNA affects the free energy of the transition such that the B form is favored by as much as 10 kcal/mol. The presence of the marked structural differences between B- and A-DNA have consequence for understanding thermal and conduction properties of composites of nanotubes and DNA used in biomaterials applications. Also, I will present a methodology that uses molecular-dynamics derived parameters to scale up the dynamics of DNA molecules on the micrometer-microsecond scale via Kirchhoff elasticity theory for elastic rods.

Resume : Biological and bioinspired units such as protein micelles, lipid vesicles or peptide macromolecules offer manifold functionalities at high specificity. Research in life and materials sciences develops methods to make these functional bio units accessible for next generation devices for e.g. sensing, molecular electronics or bio photonics. Most bio units form in liquid suspensions. This is a suitable environment for e.g. drug delivery systems, however is a drawback for the integration of bio units into electronic devices. Facing this challenge, here we present an approach for the site-selective deposition of single functional bio units, i.e. protein micelles and DNA origamis, into ordered arrays on large areas of solid surfaces. In particular, we use nanosphere lithography as a bottom-up approach for the nanopatterning of different material surfaces. We create nanohole arrays, i.e. ordered cylindrical holes in thin films, exhibiting a surface topography along with a local material contrast. We use these nanoholes as templates for the site-selective deposition of casein micelles. We take advantage of the templates chemical contrast. We locally perform an enzyme mediated autodeposition and create nanostructured biocoatings. In another example, we site-selectively adsorb DNA origamis inside the nanoholes. These origamis are for instance suitable transporters for quantum dots enabling bioinspired next generation nanoelectronics.

Resume : The recent rise in life expectancy has led an increase in the number of cases of neurological diseases such as Alzheimer?s, Parkinson?s and macular degeneration. Traditional therapeutic approaches are ineffective as regeneration is limited in the Central Nervous System (CNS). Neural prosthetics and stem cell therapy present exciting solutions for enabling the function of the brain to be restored. Implanted materials for neuronal prosthetics must have outstanding electrical properties whilst being inert and biocompatible. Neural stem cells (NSCs) have great potential for inducing repair in damaged areas of the central nervous system (CNS). NSCs have the ability to self-renew, but also are able to differentiate into the main cells in the CNS: neurons, oligodendrocytes and astrocytes. Understanding the differentiation into these cell lineages is critical for regenerative therapy treatment of such for mentioned neurological diseases as detailed knowledge of how these specific cells are affected by disease is vital (1). In order to utilise the potential of stem cells in the field of regenerative medicine, it is essential that we are able to isolate the cells from their natural setting, propagate the cells in culture, and introduce the cells to a foreign environment (2). Given the outstanding bioactivity shown by nanodiamonds (NDs) towards neurons (3), the proliferation and differentiation of human NSCs (hNSCs) and adipose derived stem cells (ADSCs) and their relationship with diamond has been investigated. Here, the biocompatibility of diamond upon boron inclusion and nanostructuring has been investigated, as well as the interaction of hNSCs and ADSCs with functionalised NDs. Oxygen-NDs significantly favouring the proliferation and adhesion of hNSC over Hydrogen-NDs and the tissue culture polystyrene (TCPS) control. Contact angle and protein adsorption investigations suggesting why O-NDs are highly suited for hNSC growth. Also, ND surfaces with O and H functionalisation are shown to influence the differentiation of hNSCs in varying ways, with hNSCs fate being investigated via both inductive and spontaneous differentiation assays. 1. Lindvall O, Kokaia Z. Stem cells for the treatment of neurological disorders. Nature. Nature Publishing Group; 2006 Jun 29;441(7097):1094?6. 2. Scadden DT. The stem-cell niche as an entity of action. Nature. 2006 Jun 29;441(7097):1075?9. 3. Thalhammer A, Edgington RJ, Cingolani LA, Schoepfer R, Jackman RB. The use of nanodiamond monolayer coatings to promote the formation of functional neuronal networks. Biomaterials. 2010 Mar;31(8):2097?104.

Resume : One of the latest trends in the fields of tissue engineering as well as oncological research is the development of in vitro 3D systems mimicking tissues or organs in vitro. Indeed, there is an increasing demand for biomimetic in vitro models recapitulating the tridimensional structure and microenvironment experienced by cells in vivo. In addition to chemical and mechanical cues, some tissues are known to be regulated by endogenous bioelectrical cues. One such tissue is bone. It has also been demonstrated to exhibit piezoelectric properties in vivo. Electroactive scaffolds, based on conducting polymers, are promising candidates to address bone piezoelectricity and to enhance osteogenesis in vitro as well as in vivo. Herein, we describe the development of a biomimetic 3D model of bone tissue based on the conducting polymer PEDOT:PSS and human adipose derived stem cells. 3D electroactive porous scaffolds were produced using the ice-templating technique. Different PEDOT:PSS to Collagen ratios) were explored. Pores sizes, mechanical and impedance properties were measured as a function of scaffolds composition. Osteogenic differentiation studies were run and the different compositions were assessed for their impact on stem cells fate. SEM/FIB was used to investigate the interaction between materials and cells, highlighting an intimate contact of the cells lining the pores walls. The results obtained so far underline the usefulness of the developed porous conductive scaffolds as in vitro platforms to study hADSCs for bone tissue engineering.

Resume : Cancer cells can become resistant to chemotherapeutic drugs and pose a challenging impediment for oncologists in providing effective chemotherapy treatment. Nanomedicine may allow overcoming chemoresistance and is the focus of our investigation. Here we show the validity of nanomedicine approach for targeted chemotherapeutic cargo delivery to overcome chemoresistance in cancer cells both in vitro and in vivo. For this, we functionalise ~100 nm long porous silica nanoparticles (~20 nm diameter ordered pore structure) by conjugating anticancer drug, cytochrome c enzyme and dual-function anticancer aptamer AS1411 in single supra-assembled nanocargos. The supra-assembly on the porous silica nanostructure allows for a high loading of catalytic enzyme cytochrome c, anticancer drug and aptamer. The silica supra-assembly is characterized by transmission electron microscopy (TEM) and BET analysis. Conjugation of cargoes has been monitored at each step by UV-Vis and Fluorescence spectroscopy. Finally, the constructed supra-assembled nanocarrier tested in vitro and in vivo. A pH-responsive, intracellular theranostic cargo delivery has been achieved and the triple action of the nanocargo made an efficient killing of drug resistance colon cancer cells in vitro by supressing the P-glycoprotein (P-gp) level. The nanocargos displayed triplex therapy effects on the drug resistance cancer cells both in vitro and in vivo

Resume : Tuning nanoparticles surfaces is very important to introduce new properties to the system such as solubility, self-assembly, sensing, biocompatibility, etc.1, 2 This talk will explain the growth of silver nanoparticles in aqueous solution, without the presence of typical surfactant molecules, but under the presence of different proteins.3 The shape of the resulting silver nanoparticles could be tuned by the selection of the types of proteins. The number of accessible lysine groups was found to be mainly responsible for the anisotropy in nanoparticle formation. Viability measurements of cells exposed to protein capped spherical or prism-shaped NPs did not reveal differences between both geometries. Thus, in the case of protein protected Ag NPs, no shape-induced toxicity was found under the investigated exposure conditions.

Resume : The desire to restore the quality of life to transtibial amputees in Nigeria has been on the front burner in recent years. In this study, a home-grown nanocomposite (NC) material (multi-walled carbon nanotube reinforced natural rubber) and multiflex dynamic response 2 foot (a common foreign foot prosthetics in Nigeria) were investigated with a view to comparing their water absorption capacity, thermal stability, wear resistance and morphological properties. The inherent challenge of ensuring uniform distribution of multi-walled carbon nanotube (MWCNT) in the host matrix was addressed by the use of sodium dodecylbenzene sulfonate (C18H29NaO3S). The CNT was synthesised via catalytic chemical vapour deposition (CCVD) technique and the NC was produced using an electrically heated hydraulic press. While the initial decomposition temperatures (Tonset) of the materials show that the newly developed NC with 260.01 oC is more thermally stable than M. DR2 foot with the temperature of 238.17 oC, incorporation of MWCNTs into the unfilled NR matrix shows a significant change in Tonset. MWCNT loading was found to influence the moisture content of the reinforced matrix by about 7% with the NC being 35% more thermally stable than M. DR2 foot. SEM/EDS micrographs indicated complete embedment of MWCNTs in NR matrix thereby making it more suitable than M. DR2 foot which was inundated with cavities. While it takes both NR/MWCNT and DR2 foot 120 days to attain saturation point in water, the former is 93% more dimensionally stable than the latter and also demonstrated better resistance to wear than the latter. It can, therefore, be concluded from the foregoing that the home-grown material is to be preferred to its foreign counterpart for anthropomorphic prosthetic foot application. Keywords ? Natural rubber, NR/MWNT, TGA/DTG, SEM/EDS, Water Absorption, DR2 foot, Wear Resistance

Resume : The high hydrophobicity of fullerenes and the resulting formation of aggregates in aqueous solutions hamper the possibility of their exploitation in many technological applications. Noncovalent bioconjugation of C60 with proteins is an emerging approach for their dispersion in water. Using lysozyme and C60 as model systems and NMR chemical shift perturbation analysis, a protein-C60 binding pocket was identified unambiguously in aqueous solution [1]. Lysozyme forms a stoichiometric 1:1 adduct with C60 and conserves its tridimensional structure upon binding. Only few residues, localized in a well-defined protein binding pocket, are perturbed. AFM, cryo-TEM and high resolution X-ray powder diffraction show that the C60 dispersion is monomolecular. The adduct is biocompatible, stable in physiological and technologically-relevant environments, and easy to store. Hybridization with lysozyme preserves the photophysical and electrochemical properties of C60. Near infrared fluorimetry and EPR spin-trapping experiments show that the C60@proteins hybrids produce reactive oxygen species (ROS) following both the type I and type II mechanisms [2]. C60 shows a significant visible light-induced generation of ROS, that can be exploited in photocatalysis or photodynamic therapy. The non-covalent bioconjugation of C60 with different proteins offers a palette of carriers for fullerenes for all pH ranges.

Resume : Keynote Oral Presentation - Young Scientist Forum (VSF) Scanning tunnelling microscopy (STM) is one of the most commonly used techniques for high resolution imaging of molecules and also to measure their electrical properties at the single-molecule level[1-5]. It is a technique that has been used to control the formation of self-assembled monolayers revealing the parameters which are involved in their formation whether they are physical parameters (e.g. concentration of solutes, wettability, contact angles, superficial tension, temperature variations) or chemical parameters (e.g. molecule structure, orbital configuration). Mainly, the STM performed at the liquid/solid interface is an ideal tool to investigate self-assembly phenomena in real time imaging the molecules adsorption dynamics[2]. Recently, the STM has been used to build multicomponent molecular systems, e.g., self-assembled monolayers with more than one molecular element, vertical supramolecular structures synthetized in-situ[3,4], molecular switches[2], nanostructured and biocompatible interfaces[5]. This work reviews the methodology which is involved in the development of these systems revealing details on how to use the STM to monitor their fabrication[6]. [1] Q. Ferreira, L. Alcácer, J. Morgado, ?Stepwise Preparation and Characterization of Molecular Wires made of Zinc octaethylporphyrin complexes bridged by 4,4?-bipyridine on HOPG?, Nanotechnology, 22, 435604, 2011 [2] Q. Ferreira, A. M. Bragança, N. M. M. Moura, M. A. F. Faustino, L. Alcácer, J. Morgado, ?Dynamics of porphyrin adsorption on highly oriented pyrolytic graphite monitored by scanning tunnelling microscopy at the liquid/solid interface?, Applied Surface Science, 273, 220, 2013. [3] Q. Ferreira, A. M. Bragança, L. Alcácer, J. Morgado, ?Conductance of well-defined porphyrin self-assembled molecular wires up to 14 nm in length?, Journal of Physical Chemistry C, 118 (3), 7229 - 7234, 2014. [4] J. Oliveira, A. M. Bragança, L. Alcácer, J. Morgado, Mário A. T. Figueiredo, J. Bioucas, Q. Ferreira, Sparse-coding denoising applied to reversible conformational switching of a porphyrin self-assembled monolayer induced by scanning tunnelling microscopy: IMAGE DENOISING ALGORITHMS FOR STM IMAGES, Journal of Microscopy, DOI: 10.1111/jmi.12699, 2018 [5] A. M. Braganca, L. Alcacer, A. Ferraria, A. Rego, J. Morgado, and Q. Ferreira, Bioactive nanostructured monolayer with glucuronic acid assembled on graphite, submitted [6] C. Delfino, Q. Ferreira, Recent Advances in bottom-up self-assembled supramolecular structures built by STM, Materials, 2018, accepted.

Resume : Engineered nanoparticles (NPs) have been shown to be a promising tool for biofilm prevention and disruption. Although many studies indicate that the most likely mechanism for this action is the interference with the bacterial metabolism and cellular membranes, the role of the biofilm extracellular polymeric matrix (EPS) in the framework of the NPs-biofilm interaction is not yet fully understood, and quantitative relationships between NPs properties and EPS composition are still lacking. This contribution deals with the use of engineered fluorescent silica NPs to elucidate the role of the EPS matrix in NPs-biofilm interactions, especially how the EPS composition, density and structure affect phenomena such as NP diffusion, uptake and accumulation within the biofilm. Epoxide-engineered fluorescent silica NPs were prepared, and the epoxide moieties were used as anchoring platform for further functionalisation (modification with amine groups, PEG, aromatic or alkyl groups). These NPs were injected in biofilms grown from Pseudomonas strains and the interaction with the EPS studied through confocal microscopy, UV-vis, IR and fluorescent spectroscopy, DLS and Z-potential analysis. The results show that selective interactions with the EPS take place according to specific surface functionalisation. The outcome of this study will be useful in applications where antibiofouling technology is needed, such as the water, biomedical and food industries.

Resume : With increasing demand for the detection of delicate bio-signals for medical electronics, the Internet of Things (IoT), E-skin and flexible integrated circuit (IC) devices, an enhancement in sensitivity has become a major issue in flexible mechanosensors, however, overcoming the limited sensitivity remains problematic. Here, we introduce mechanosensors inspired by spiders having an ultrasensitivity, durability. For ultrasensitivity and durability, we considered the geometrical effects in cracks and self-healable polymers. By controlling crack depth by simple propagating process, the sensitivity of our sensor shows ~15,000 in 2% strain, which is the world best sensitivity value. Due to the high sensitivity, the signal-to-noise-ratio is 6 times higher than before, up to ~35 so that it can be used in sensing human voice clearly. Also, self-healable polymer helps to recover the crack gaps after 25,000 cycles. We introduce the possilibility of semi-permanent uses over 1,000,000 cycles in our sensors. The spider inspired sensory system with high sensitivity and durability would provide versatile novel applications such as E-skins, devices for medical applications, and IoT applications etc.

Resume : Poly(2-methoxyethyl acrylate) (PMEA) shows excellent blood compatibility due to the existence of intermediate water (1). Small amount of amino groups was found to change the hydration structure of 2-hydroxyethyl methacrylate when combining in a copolymer structure, which additionally decreased the interactions with lymphocytes (2). Here we exploit another possibility to manipulate the surface hydration structure of PMEA by incorporation of small amount of other than nitrogen - the hydrophobic fluorine groups in MEA polymers using Atom Transfer Radical Polymerization and the (macro) initiator concept (3). Focusing on the difference in mobility, two kind of fluorinated MEA polymers were synthesized using 2,2,3,3,4,4,5,5,6,6,7,7,8,8-pentadecafluoro-1-octanol (F15) and poly(2,2,2-trifluoroethyl methacrylate) (P3FM) (macro) initiators appearing liquid and solid at room temperature, respectively. The fibrinogen adsorption of the two varieties of fluorinated MEA polymers was different, that could not been explained only by the bulk hydration structure. Contact angle and AFM measurements reveal that the F15-PMEA reorients in water easily to the surface as compared to the P3FM-b-PMEA which reorientation was suppressed by the small solid fluorinated P3FM block. These findings illustrate, that in order to make a better bio-inert material, the chains containing sufficient intermediate water need to be efficiently oriented to the water surface. References: (1). M. Tanaka, A. Mochizuki. J. Biomed. Mater. Res. 2004, 68A, 684-695. (2). T. Tsuruta. J. Biomater Sc. 2010, 21, 1831-1848. (3). K. Jankova, X. Chen, J. Kops, W. Batsberg. Macromolecules 1998, 31, 538-541.

Resume : Electrospinning has emerged as a versatile, cost effective and reliable technique for fabrication of micro/nano fibers and particles with a strict regulation of solution parameters. However, there is minimal evidence till date of any successful dual structured particle on fiber ?Sandwich?drug delivery system obtained from electrospinning that could provide controlled release of two or more different drugs. In this study we have successfully fabricated a dual structured electrospun fibrous - micro particle system through electrospinning for delivery of a hydrophilic and a hydrophobic drug simultaneously. A blend of two FDA approved biodegradable polymers was used as the electrospinning solution. Preliminary work focused on optimization of the processing parameters to arrive at 5-8?m porous particles that were electro-sprayed on a fibrous mesh network of PLGA-PCL fibres. The particles were further protected by another fibrous mesh on the top to prevent its loss during post fabrication, thereby the entire system resembling a sandwich. Characterization of the scaffold was carried out using Scanning Electron Microscopy, Rheometer and Fourier transform Infrared Spectroscopy. The hydrophobic Dexamethasone (Dex) was loaded into the fibers while Ascorbic acid (AA) was the hydrophilic biomolecule loaded into the particles. The release profiles of these drugs were quantified with the help of High Performance Liquid Chromatography (HPLC) which enabled the detection of Dex at 246nm and AA at 262 nm respectively. These scaffolds were used to study the differentiation capabilities of mesenchymal stem cells (MSCs) into osteo and chondro lineage. Future studies are directed towards understanding the influence of the carrier morphology and the synergistic effect of multiple biomolecules will be explored in depth for musculo-skeletal tissue engineering.

Resume : Hydrogen peroxide (H2O2) is a strong oxidizing compound. It is involved in signalling and defence responses in biological systems, where enzymes (oxidases) reduce it to H2O through different pathways. In food and clinical analysis, concentrations of organic molecules are quantified by measuring the H2O2 produced by the coupled activity of oxidases. Although many methods are available for H2O2 detection, they are not suitable for industrial application. Electrochemical techniques, such as voltammetry, are advantageous for low level H2O2 detection in industrial applications because they are simple, fast and sensitive. Since commonly used electrodes are subject to non-linearity in their response, poisoning, and interference with collateral reactions when complex matrices are involved, we here propose an alternative sensing system. Cerium oxide (CeO2) nanoparticles show oxidase-like activity due to their capacity to change electronic configuration and exhibit oxygen vacancies, or defects, in the lattice structure. We conducted an electrochemical, morphological and spectroscopic study aimed at investigating the performance of polycrystalline CeO2 thin layers on Glassy Carbon electrodes as redox sensors, detecting micromolar concentrations of H2O2 in Phosphate Buffer Solution. Surface characterization was performed by SEM and AFM imaging before and after the electrochemical measurements, while system stability was investigated by Synchrotron Radiation Photoelectron Spectroscopy.

Resume : Poly(3,4-ethylenedioxythiophene) (PEDOT) materials was considered as star materials for fabricating bioelectronic devices 1-4 due to its excellent electrical conductivity and electrochemical stability5,6. Decoration with functional groups and introducing nanostructures would enhance the electric and biological properties of PEDOT materials, which could expand the potential applications in the fields of bioelectronics. Therefore, it was important to design simple but controllable nano-assembling methods for functionalized PEDOT materials. Compared to traditional template assembling, template-free method had the beneficial of easy fabrication in large scale and integrated preservation of achieved nanostructures, which would make it possible for fabricating nano electronic devices in wafer-scale. Several methods had been reported to realize nano assembling PEDOT materials, however, few of them could realize fabricating different types of nano-morphologies for various functionalized EDOT monomers with one simple method. Herein, we proposed template-free method for fabricating various functionalized PEDOT thin films with nanodot and nanotube morphologies via electrochemical depositing. The morphology parameters could be tuned including diameter, length and density. Glancing incident wide angle XRD was measured to testify the difference of crystal orientation for the achieved nanodot and nanotube thin films. Meanwhile, possible formation reason and process of nanodot and nanotube morphologies were proposed. Besides, simulation tests were carried out and verified the beneficial of the PEDOT and functionalized PEDOT thin films with nanotube morphology using in bioelectronic devices. We hoped the achieved template-free assembling method could be expanded to other kinds of bio-functionalized PEDOT materials and designed PEDOT thin films with proper bio-functional groups and 3D nanomorphologies based on the requirements in practical application of bioelectronic devices. Reference: 1. Khodagholy, D.; Rivnay, J.; Sessolo, M.; Gurfinkel, M.; Leleux, P.; Jimison, L. H.; Stavrinidou, E.; Herve, T.; Sanaur, S.; Owens, R. M.; Malliaras, G. G. Nat Commun 2013, 4, 2133. 2. Rivnay, J.; Inal, S.; Collins, B. A.; Sessolo, M.; Stavrinidou, E.; Strakosas, X.; Tassone, C.; Delongchamp, D. M.; Malliaras, G. G. Nat. Commun. 2016, 7, 11287. 3. Williamson, A.; Rivnay, J.; Kergoat, L.; Jonsson, A.; Inal, S.; Uguz, I.; Ferro, M.; Ivanov, A.; Sjostrom, T. A.; Simon, D. T.; Berggren, M.; Malliaras, G. G.; Bernard, C. Advanced materials 2015, 27, 3138. 4. Williamson, A.; Ferro, M.; Leleux, P.; Ismailova, E.; Kaszas, A.; Doublet, T.; Quilichini, P.; Rivnay, J.; Rozsa, B.; Katona, G.; Bernard, C.; Malliaras, G. G. Advanced materials 2015. 5. Sotzing, G. A.; Briglin, S. M.; Grubbs, R. H.; Lewis, N. S. Analytical chemistry 2000, 72, 3181. 6. Jalili, R.; Razal, J. M.; Innis, P. C.; Wallace, G. G. Advanced Functional Materials 2011, 21, 3363.

Resume : Silkworm silk and spider silk with outstanding properties have unique hierarchical structures at mesoscale. As the basic building block of the hierarchical structure, silk nanofibrils (SNF) is the key unit for the formation of high performance silk based materials. Conventional methods to prepare SNFs have some limitations, such as nanofiber aggregation/inadequate dissociation, low yield, toxic solvent, imperfect building model of silk hierarchical structure etc. To fabricate stable SNFs suspension efficiently, a novel solution system was used to prepare SNFs. Transmission electron microscope, atomic force microscope, and synchrotron radiation small angle X-ray scattering were applied to confirm the size of SNFs accurately. Moreover, a SNF ultra thin film with good flexibility, high transmittance, biocompatibility and biodegradability were fabricated via vacuum filtration from the SNFs suspension. The film may have potential application for biosensing devices, optics, photonics or tissue engineering.

Resume : High temperature frying, broiling, grilling and especially smoking protein providing food products, such as meat, fish, poultry, eggs, and cheese generates in these products toxins, namely heteroaromatic compounds, amines, nitrosamines etc. Continuous exposure to low doses of these toxins causes several chronic diseases, serious hormonal dysfunctions, and cancer. One of the trace amines, tyramine, may also cause unwanted interactions with antidepressant monoamine oxidase (MAO) inhibitors causing so called “Cheese reaction”. Currently used procedures for determination of these toxins in food matrices are either expensive or tedious and time-consuming. Therefore, fast, inexpensive, simple, and reliable determination procedures, without need of separation of these toxins, in the protein food matrices are in demand. Molecularly imprinted polymers (MIPs) are excellent example of bio-mimicking recognition materials. Therefore, they have found numerous applications in selective chemosensing. Within this project we have applied tyramine imprinted polythiophene films as a selective recognition units for devising electrochemical sensors. Tyramine-MIP film coated electrodes showed good sensitivity and selectivity with respect to tyramine. The linear dynamic response range of the devised chemosensor was from 260 µM to 2,6 mM tyramine and imprinting factor was IF = 3.

Resume : Diseases typically result from the accumulation of substances that disrupt the normal operation of cells. The introduction of enzymes, catalytic protein molecules essential for normal biological function, which act on such substances into affected tissues provides an attractive alternative for curing such diseases. Silicon nanocrystals, owing to their limited toxicity and photodynamics, offer a potentially safer and more efficient bioimaging platform compared to status quo organic dyes. Thus, a hybrid material consisting of enzymes that have been interfaced with silicon nanocrystals could offer simultaneous imaging and therapy. This study reports, for the first time, methods for the preparation of enzyme-conjugated silicon nanocrystals from native enzymes and acid or alkene-terminated silicon nanocrystals through the amide coupling and thiol-ene reactions, respectively. Model enzymes, glucose oxidase and lactase were successfully immobilized on silicon nanocrystals as confirmed by Fourier Transform Infrared Spectroscopy and X-ray Photoelectron Spectroscopy. Moreover, single reaction and cascade kinetic assays confirm that the conjugated enzymes retain their catalytic activity. The hybrids manifested either excellent solubility or good dispersibility in buffer, and were photostable, exhibiting bright orange photoluminescence even after more than a month of dispersion in an aqueous medium. The methods reported herein are general and can be used for the preparation of bioinorganic silicon-based hybrids that can be employed in personalized medicine for targeting and potentially treating diseases like cancer and other metabolic disorders.

Resume : Advanced affinity tools for cell imaging are of particular interest as they can detect, localize and quantify molecular targets. Aberrant gly-cosylation sites and deregulated expression of growth factor receptors are promising bi-omarkers of many human diseases, most no-tably cancer. However, targeting these bi-omarkers is often challenging due to a lack of receptor molecules. Molecularly imprinted pol-ymers (MIPs) are tailor-made synthetic recep-tors (antibody mimics), able to specifically rec-ognize target molecules. They are synthesized by co-polymerizing functional and cross-linking monomers in the presence of a molecular template, resulting in the formation of binding sites with affinities and specificities compara-ble to those of natural antibodies [1]. Herein, we demonstrate biotargeting and bi-oimaging with fluorescently labeled MIPs on two different cancer biomarkers: hyaluronan and a growth factor receptor protein. MIPs were synthesized using a solid-phase synthe-sis approach in which an epitope of the bi-omarkers was immobilized on glass beads (as solid support) via click chemistry. This configu-ration allows an oriented immobilization of the template upon which thermoresponsive MIP nanoparticles were synthesized. The binding sites of the resulting MIPs all have the same orientation, thus MIPs synthesized by the sol-id-phase approach can be considered analo-gous to monoclonal antibodies [2-3]. Hyaluronan imaging was achieved by applying rhodamine-doped MIPs specific for glucuronic acid (an epitope of hyaluronan) on fixed hu-man keratinocytes [4]. Hyaluronan is com-posed of alternating units of D-glucuronic acid (GlcA) and N-acetyl-D-glucosamine. Thus, azide functionalized-glucuronic acid was im-mobilized on glass beads bearing alkyne groups. MIP-GlcA (70 nm) as water soluble particles were found to bind selectively extra-cellular, intracellular and nuclear hyaluronan, as imaged by epifluorescence and confocal microscopies. The specificity of bind-ing was verified with a non-imprinted control polymer and by comparing the staining with a hyaluronan binding protein. For bioimaging the growth factor receptor membrane protein, a short peptide (terminal alkyne functionalized) was selected as epitope for immobilization on azide-modified glass beads. The MIP nanoparticles (50 nm) specifi-cally recognized both the template peptide and the whole protein. Cell imaging studies with fluorescent dye-doped MIPs were per-formed. RFERENCES: [1] K. Haupt, A. V. Linares, M. Bompart and B. Tse Sum Bui, Top. Curr. Chem. 325 (2012), 1-28 [2] S. Ambrosini, S. Beyazit, K. Haupt and B. Tse Sum Bui, Chem. Commun. 49 (2013), 6746-6748 [3] J. Xu, P.X. Medina-Rangel, K. Haupt and B. Tse Sum Bui, Methods Enzymol. 590 (2017), 115-141 [4] M. Panagiotopoulou, S. Kunath, P. X. Medina-Rangel, K. Haupt and B. Tse Sum Bui, Biosens. Bioelectron. 88 (2017), 85-93

Resume : Despite strict compliance with the standards of ISO 10993 with respect to medical implants, their widespread use proves a certain level of risk of implant failure. Among the main problems regarding the use of implants in clinical practice microbial biofilm-related infections and an insufficient integration of implant with tissues are serious obstacles. Currently available biomaterials are relatively well-suited to strength requirements, therefore, the methods of surface modification of implants are used to improve their properties and minimize possible negative outcomes of the use of biomaterials. Moreover, changes and proper control of process parameters allow to get surfaces better adapted to the needs and expectations of personalized medicine. Parameters of anodization process can be easily altered and well controlled, thus leading to the tailoring of properties of the resulting oxides. In the performed research, the anodic oxidation was used for creation of titanium dioxide films (TiO2) on surfaces of titanium alloy Ti6Al4V samples. The coatings differed from each other in the surface structure. Four different types of surface textures were obtained (smooth, porous, nanotubular and nanorough). Additionally, each type of the coating was doped with antimicrobial agents – copper and zinc. Presence of dopants decreased the number of E.coli cells which adhered to the examined surfaces. Biological evaluation concerning the live/dead test for osteoblasts indicated that the created layers were not cytotoxic. The presented results prove a real possibility of the use of titanium dioxide films doped with copper and zinc for medical applications. Acknowledgement: The work was supported by the Polish National Center for Research and Development project POIR.04.01.04-00-0058/17-00.

Resume : We will report the synthesis of triple labeled Mesoporous Silica Nanoparticles (FORMSN) by encapsulates 3 kind of dyes: Fluorescein isothiocyanate(FITC, pKa=6.7) and oregon green succinimidyl ester (OG, pKa=4.8) as pH sensitive dyes and rhodamine isothiocyanate (RITC) as a reference dye, which have broad range pH detection. FORMSN have also been decorated with lysosomal sorting peptide (YXXɸ) for targeting lysosome. The 3D single particle movement based on pH detection were demonstrated by using this FORMSN. We too found out that with the peptide conjugation increased the number accumulation of particle in lysosomal compartment. Moreover, we noticed 3 different patterns of the pH changing to the time of FORMSN-peptide during traffic routes: acidification, basification, and stabilization.

Resume : Multilayer graphene (MLG) has been actively investigated because of its high electrical and thermal conductivities. Because graphene has a unique two-dimensional structure, its characteristics are anisotropic. Therefore, large-grained highly oriented MLG on insulators is highly desirable. Metal-induced layer exchange (MILE) is a promising technique allowing for large-grained, highly-oriented Ge and Si on glass [1]. In this study, we applied MILE to amorphous carbon (a-C) and fabricated high-quality MLG at as low as 500 °C. Ni and a-C thin films (each 50 nm thick) were sequentially prepared on glass using magnetron sputtering. Samples were annealed at 500 °C for 50 h. The Ni layers were then etched away. The Raman spectra of back side of the samples have sharp D, G, and 2D peaks corresponding to MLG, indicating the layer exchange between the C and Ni layers [2,3]. The cross-sectional TEM analyses showed that the layers of a-C and Ni were exchanged and {002} oriented MLG formed on glass substrate. The grain size was approximately several hundred nm. After the removal of the Ni layers, the MLG covered the entire substrate. The electrical conductivity was approximately 400 Scm< sup>-1< /sup>. The uniformity and electrical conductivity are the highest level among the MLG directly formed on glass at low temperature. [1] Toko et al., APL. 104, 022106 (2014). [2] Murata et al., APL. 110, 033108 (2017). [3] Murata et al., APL. 111, 243104 (2017). (Highlighted in Nature INDEX.)

Resume : A cicada wing has a biocidal feature of rupturing the membrane of cells, while the cactus spine can transmit a water drop to the stem of the plant. Both of these properties have evolved from their respective unique structures in nature. Here, we endeavor to develop geometry-controllable polymer nanohairs that mimic the cicada?s wing-like vertical hairs and the cactus spine-like stooped hairs, and to quantitatively characterize the cell migration behavior of the hairy structures. It was found that the neuroblastoma cells are highly sensitive to the variation of surfaces: flat, vertical, and stooped nanohairs (100 nm diameter and 900 nm height). The cells on the flat structures showed random movement while the cells on the vertical hairs showed significantly decreased proliferation. It was also found that the behavior of cells cultured on stooped nanohairs is strongly influenced by the direction of the stooped pattern of hairs when we quantitatively measured the migration of cells on flat, vertical, and stooped structures. Cells on the stooped structure showed higher forward migration preference compared to that of the other structures. Furthermore, we found that these cellular behaviors on the different patterns of nanohairs were affected by intracellular actin flament change. Consistent with these results, the vertical and stooped structures can facilitate the control of cell viability and guide directional migration for biomedical applications such as organogenesis.

Resume : Cancer is the second leading cause of death in the world. So its early detection and therapy is very important. There are many different ways such as blood tests, X-rays, (contrast) CT scans and endoscopy to diagnose cancer that are so expensive and time-consuming. The effective and rapid detection of cancer cells at various stages are the challenges of diagnosis of cancer. Herein, we introduce a lab-on-phone device that has the ability to detect vascular endothelial growth factor (VEGF) angiogenesis on the surface of the cancerous cells. The benefits of this approach are being easy to operate, portable, low cost, reliable and fast. This method is based on the localized surface plasmon resonance (LSPR) property of gold nanoparticles (GNPs) which play the key role in this measurement. In this work, a reference solution was made that acts as a probe for detection of VEGF. Gold nanoparticles were linked to Bevacizumab (Avastin) antibody (BAB) by mercaptopropionic acid (MPA) and EDC/NHS. Then, GNPs were agglomerated due to BAB connection to VEGF on the surface of cancerous cell. The color of solution changed from red to purple due to GNPs agglomerations. This color change was analyzed by taking a photo by the phone?s camera using a generated software application which can process the photos and convert this change to the analyte concentration.

Resume : The epidermal equivalents are used as an alternative animal test of permeation and toxicity screening of chemicals. In this study, we demonstrate stereolithography three dimensional (3D) printing for the mimicry of the epidermal equivalents. In comparison with traditional methods for constructing epidermal equivalents, it offers advantages in terms of cell laden retention, reproducibility and high culture throughput. Human keratinocytes are incubated on a silk fibroin-PEG hydrogel scaffold containing fibroblasts and also grown at the air-liquid interface, which allows the enhanced maturation and stratification. In the presentation, the detailed process of purification of silk fibroin from the cocoon, the synthesis of acrylated-PEG, 3D printing of bioinks, and the critical parameters of the cell culture will be discussed.

Resume : The epidermal equivalents are used as an alternative animal test of permeation and toxicity screening of chemicals. In this study, we demonstrate stereolithography (SLA) three dimensional (3D) printing for the mimicry of the epidermal equivalents. In comparison with traditional methods for constructing epidermal equivalents, it offers advantages in terms of cell laden retention, reproducibility and high culture throughput. Primary human keratinocytes are incubated on a silk fibroin-PEG hydrogel scaffold containing fibroblasts and also grown at the air-liquid interface, which allows the enhanced maturation and stratification. In the presentation, the detailed process of purification of silk fibroin from the cocoon, the synthesis of acrylated-PEG, 3D printing of bioinks, and the critical parameters of the cell culture will be provided. Keyword Epidermal equivalents, Silk, PEG hydrogel, Alternative animal test, Stereolithography (SLA)

Resume : The development of new drug delivery (DD) systems able to release the drugs during prolonged periods has been receiving greater attention in recent years due to the ability to use these systems to treat diseases without human intervention. Mainly in ocular diseases with most of treatments consisting in applying eye drops which has a poor patient compliance. We are developing DD multilayers films able to release an ocular drug used in glaucoma treatment which can release the drug during a month and at specific periods of time. Biocompatible films composed of brimonidine encapsulated in ?-cyclodextrin alternated with monolayers of a hydrossoluble polymer (poly (?-amino ester)) and/or graphene oxide are able to release a precise amount of drug for a month. The ?lms growth and the pharmacokinetics were monitored by ultraviolet-visible spectroscopy, quartz crystal microbalance and atomic force microscopy. The obtained results showed that the ?lms are stable and drug release can be controlled by the presence of the hydrossoluble polymer and the graphene oxide. In particular, it was observed that graphene oxide delays signi?cantly the brimonidine release enabling precisely control the amount of drug delivered. This work contributed for new developments in DD ?lms that can be used in glaucoma treatment or adapted to other DD systems with other types of drugs. Acknowledgements We thank FCT-Portugal, under the project UID/EEA/50008/2013 for financial support

Resume : Microbial Fuel Cell (MFC) is a sustainable energy transducer, that directly coverts organic matter into electrical energy. It shows promise in both wastewater treatment and bio-energy production. A bio film of photosynthetic green alga Chlamydomonas sp. TRC-1 deposited on fluorine tin oxide (FTO) electrodes was investigated for its ability to generate power. Cyclic voltammetry (CV) scans recorded a sharp anodic and cathodic peak with a potential difference ?V= 0.239 V. A peak power output of 10.02 mW/m2 was observed with a current density of 27A/m2. The algal biofilm applied in MFC improved the physicochemical parameters of the wastewater, significantly reducing the chemical oxygen demand (COD: 77.1%), total dissolved solids (TDS: 82.1%) and total suspended solids (TSS: 87.4%). The study not only offers an economically and eco-friendly solution to successful power generation but also contributes towards waste water treatment and biofuel production.

Resume : Self-assembled phthalocyanines (Pcs) monolayers have been studied with scanning tunnelling microscopy (STM) due to the organized 2D arrays that they form with potential applications in functional organic devices. The metal central region gives them important electronic, optical and optoelectronic properties and the possibility to extend the 2D monolayer into three dimensions through vertical reaction chemistry [1,2]. Synthetized metal Pcs [3,4] were assembled in graphite and followed by STM. High resolution images were obtained during the formation of the monolayer using a method previously applied in porphyrin monolayers [1,2,5,6]. The molecules were added to graphite trough an interface with tetradecane and the monolayer formation was followed in real time using molecular resolution images obtained by STM. Differential functional theory simulations were made to support the STM results. . [1] Q. Ferreira et al, J. Phys. Chem C, 2014 [2] Q. Ferreira et al, Nanot., 2011 [3] A. Sastre et al, Sust. Energy Fuels, 2017 [4] A. Sastre et al, J. Phys. Chem. C, 2016 [5] Q. Ferreira et al, Applied Surface Science, 273, 220, 2013 [6] Q. Ferreira et al, J. of Microscopy, 2018 Acknowledgements We thank FCT-Portugal, under the project UID/EEA/50008/2013 for financial support

Resume : Unwanted biofilm formation is currently one of the most problematic areas in the biomedical and water treatment fields. In the biofilm phenotype, a self-produced matrix of biological components (EPS ? Extracellular Polymeric Substances) acts as a chemical and physical barrier for the use of conventional biocides. Despite the extensive research of antimicrobial nanoparticles (NPs) against planktonic bacteria, information on how NPs interact and diffuse into the matrix is still a relatively unknown field, and the full potential of nanotechnology against resistant biofilms is yet to be explored. In this study, silica nanoparticles of different sizes and functionalizations (amine, carboxylic acid) were synthesized using the classic Stober and the microemulsion methods while also being labelled with fluorescent tags. EPS-NP interactions using Pseudomonas biofilms were assessed via studies using Confocal Laser Scanning Microscopy (CLSM), Dynamic Light Scattering (DLS) and Zeta potential measurements. Attachment and penetration of nanoparticles into the biofilm as well as in vitro interactions using extracted EPS were shown to be dependent on the NPs? charge and size. The preliminary results obtained on these systems introduce basic knowledge on biofilm interactions with designed nanoparticles and begin to pave the way towards the development of antimicrobial nanoparticles with activity against bacterial biofilms.

Resume : Hydrogen peroxide (H2O2) is a strong oxidizing compound. It is involved in signalling and defence responses in biological systems, where enzymes (oxidases) reduce it to H2O through different pathways. In food and clinical analysis, concentrations of organic molecules are quantified by measuring the H2O2 produced by the coupled activity of oxidases. Although many methods are available for H2O2 detection, they are not suitable for industrial application. Electrochemical techniques, such as voltammetry, are advantageous for low level H2O2 detection in industrial applications because they are simple, fast and sensitive. Since commonly used electrodes are subject to non-linearity in their response, poisoning, and interference with collateral reactions when complex matrices are involved, we here propose an alternative sensing system. Cerium oxide (CeO2) nanoparticles show oxidase-like activity due to their capacity to change electronic configuration and exhibit oxygen vacancies, or defects, in the lattice structure. We conducted an electrochemical, morphological and spectroscopic study aimed at investigating the performance of polycrystalline CeO2 thin layers on Glassy Carbon electrodes as redox sensors, detecting micromolar concentrations of H2O2 in Phosphate Buffer Solution. Surface characterization was performed by SEM and AFM imaging before and after the electrochemical measurements, while system stability was investigated by Synchrotron Radiation Photoelectron Spectroscopy.

Resume : The epidermal equivalents are used as an alternative animal test of permeation and toxicity screening of chemicals. In this study, we demonstrate stereolithography three dimensional (3D) printing for the mimicry of the epidermal equivalents. In comparison with traditional methods for constructing epidermal equivalents, it offers advantages in terms of cell laden retention, reproducibility and high culture throughput. Human keratinocytes are incubated on a silk fibroin-PEG hydrogel scaffold containing fibroblasts and also grown at the air-liquid interface, which allows the enhanced maturation and stratification. In the presentation, the detailed process of purification of silk fibroin from the cocoon, the synthesis of acrylated-PEG, 3D printing of bioinks, and the critical parameters of the cell culture will be discussed.

Resume : The demand for new functional materials dedicated for sensors and various optical devices is increasing and consequently novel and compatible high throughput micro lithography techniques are emerging. Direct laser interference patterning (DLIP) is an example of fast fabrication method, capable to impose periodic patterns in practically any material upon selection of proper ablation parameters. Ultrashort pulse irradiation can melt and therefore change structure and linear dimensions of nanoparticles. Silver nanoparticles have attracted considerable amount of interest due to their plasmonic properties and wide range of applications, among which are antibacterial coatings and various sensors. However, silver is a fast oxidising metal and thus requires passivation. One of the possible ways to achieve passivation is embedding silver nanoparticles in passivating matrix, for example diamond-like carbon. In this work, we present Yb:KGW femtosecond laser two second harmonic beams interference ablation of diamond like carbon thin films doped with silver nanoparticles. We investigate the influence on nanoparticle size distributions and one-dimensional periodic structures based on applied number of laser pulses and laser fluence. The DLIP effects are compared for nanocomposites with two different silver contents as well as pure silver and pure diamond-like carbon thin films. It was obtained that existence of silver nanoparticles in thin films lowers the ablation threshold, due to presence of localised surface plasmon absorption.

Resume : Diseases typically result from the accumulation of substances that disrupt the normal operation of cells. The introduction of enzymes, catalytic protein molecules essential for normal biological function, which act on such substances into affected tissues provides an attractive alternative for curing such diseases. Silicon nanocrystals, owing to their limited toxicity and photodynamics, offer a potentially safer and more efficient bioimaging platform compared to status quo organic dyes. Thus, a hybrid material consisting of enzymes that have been interfaced with silicon nanocrystals could offer simultaneous imaging and therapy. This study reports, for the first time, methods for the preparation of enzyme-conjugated silicon nanocrystals from native enzymes and acid or alkene-terminated silicon nanocrystals through the amide coupling and thiol-ene reactions, respectively. Model enzymes, glucose oxidase and lactase were successfully immobilized on silicon nanocrystals as confirmed by Fourier Transform Infrared Spectroscopy and X-ray Photoelectron Spectroscopy. Moreover, single reaction and cascade kinetic assays confirm that the conjugated enzymes retain their catalytic activity. The hybrids manifested either excellent solubility or good dispersibility in buffer, and were photostable, exhibiting bright orange photoluminescence even after more than a month of dispersion in an aqueous medium. The methods reported herein are general and can be used for the preparation of bioinorganic silicon-based hybrids that can be employed in personalized medicine for targeting and potentially treating diseases like cancer and other metabolic disorders.

Resume : Textile based optoelectronic devices have been widely investigated for wearable device application because of its wearability, lightweight, ease of processing, and low-cost production. Particularly, textile based organic photovoltaics (OPVs) have shown promising results. However, the photovoltaic performance of textile OPV is hindered by the non-uniform coating of conductive fibers and high sheet resistance of electrode. In this work, highly flexible, transparent hybrid conducting electrodes (TCE) were fabricated via embedding silver sub-electrodes/conductive polymers into an UV-curable polymer. The photovoltaic performance with active layer consisting of PTB7-Th:PC71BM shows a relatively high power conversion efficiency (PCE) of ~ 5% with an active area of 0.3 cm2. To prepare functional textile OPVs, multiple devices were weaved and integrated into the fiber thread with various aperture area. An output voltage of > 7 V was achieved by connecting seven devices in series. Additionally, flexible supercapacitor (SCs) were integrated with solar cells as textile based power pack. The SCs were fabricated based on PEDOT:PSS electrodes sandwiched between gel electrolyte in symmetrical device structure. An areal capacitance of >3.8 mF/cm2 was successfully achieved for the SC device. The multifunctional textile OPVs and SCs fabricated herein exhibits underlying potential for next-generation optoelectronics application.

Resume : Colorimetric detection is a widly used method for detection of analytes in different solutions. There are many instruments performing similar procedures but that are expensive and complex. Herein, we introduce a lab-on-phone device that can detect and analyze even slight color changes that are not visible for the naked eyes. The benefits of this device are being easy to oprate, portable, user friendly, low cost and fast. In our work, gold nanoparticles are playing the key role in detection of the analytes. They simly convert the analyte’s concentration to a measurable color change. This lab-on-phone device consists of lightweight opto-mechanical attachments which are wirelessly connected to a smart phone. The main active part of the opto- mechanical of device has been formed from four LEDs in blue, red, green and UV which can be used to assay different types of analytes with different absorbtions. The light absorbtion of an analyte causes a color change. This color change was analyzed by taking a photo by the phone’s camera using a generated software application which can process this photo and convert its color change to the analyte concentration.

Resume : Microbial Fuel Cell (MFC) is a sustainable energy transducer, that directly coverts organic matter into electrical energy. It shows promise in both wastewater treatment and bio-energy production. A bio film of photosynthetic green alga Chlamydomonas sp. TRC-1 deposited on fluorine tin oxide (FTO) electrodes was investigated for its ability to generate power. Cyclic voltammetry (CV) scans recorded a sharp anodic and cathodic peak with a potential difference ∆V= 0.239 V. A peak power output of 10.02 mW/m2 was observed with a current density of 27A/m2. The algal biofilm applied in MFC improved the physicochemical parameters of the wastewater, significantly reducing the chemical oxygen demand (COD: 77.1%), total dissolved solids (TDS: 82.1%) and total suspended solids (TSS: 87.4%). The study not only offers an economically and eco-friendly solution to successful power generation but also contributes towards waste water treatment and biofuel production.

Resume : A typical method to increase photoluminescence efficiency is shell formation on quantum dots (QDs) cores for removing surface trap sites. However, insulating property of QDs shell hinders efficient hole and electron injection between shell-coated QD layer and adjacent transport layers leading to decreased device efficiency when the QD layer is applied in optoelectronics. To solve this issue, thermal annealing is usually adapted to increase photoluminescence efficiency by controlling optimum amount of defect sites. On the other hands, relative high temperature used during thermal annealing may cause deterioration of interface between core and shell of QDs. To avoid the deterioration of the interface, Intense pulse light(IPL) annealing can be introduced as an alternative to thermal annealing as it almost instantaneously supplies heat within very short time and brings enough energy to QDs. In this study, we aim to maximize the photoluminescence efficiency of the QDs by controlling the defect sites of the core through a IPL annealing technique while maintaining the interface between core and shell of QDs intact. IPL annealing technique is possible to efficiently remove the defect sites by applying optimum amount of heat to quantum dots core using very short pulse. It is because IPL annealing can avoid over heating on QDs very easily by controlling pulse width. We have observed considerable improvement of quantum efficiency of the QDs by using the afore-mentioned IPL annealing process.

Resume : The long-term integration of the implants with human body depends on the chemical and physical surface properties of the substrates. Therefore, surface engineering of biomedical materials, is consider to be major technique for improving processes on the implant/body interface. A technique that can modify the titanium possessing initial, not flat topography is Direct Laser Interference Lithography (DLIL), without the deformation thereof. It can generate complex types of texture with high resolution, resulting in micrometer size patterns. Expect for modifying the surface appearance, laser irradiation, with the specific thermal characteristic, can generate appropriate microstructures below the free surface including nanocrystalline or metastable phases. The dissolution of the light elements, such as oxygen or nitrogen, strongly depends on the interactions between the laser, substrate and the atmosphere. The mentioned effects are especially important in the case of the processes limited to the substrate surface. This is of great importance in biomedical applications, where the assessment of various processes such as, protein adsorption, cell adhesion or corrosion resistance is not limited to topographical parameters but includes the formation of oxide layers and microstructural changes. Therefore, this study deals with a careful cross-section microstructural analysis by combining complementary methods such as FIB, SEM, EDS and STEM on different scales to elucidate the microstructure and surface chemistry after laser- patterning and the correlation with phase composition. The gained results are important with regard to the evaluation of laser patterned surfaces in biomedical applications. The description of the microstructure and chemical composition of the DLIL modified titanium samples, can be useful when understanding and characterizing the processes on the bone/implant interface. This research was financially supported by The National Science Centre Poland under Grant no. 2016/23/N/ST8/02044.

Resume : A cicada wing has a biocidal feature of rupturing the membrane of cells, while the cactus spine can transmit a water drop to the stem of the plant. Both of these properties have evolved from their respective unique structures in nature. Here, we endeavor to develop geometry-controllable polymer nanohairs that mimic the cicada‘s wing-like vertical hairs and the cactus spine-like stooped hairs, and to quantitatively characterize the cell migration behavior of the hairy structures. It was found that the neuroblastoma cells are highly sensitive to the variation of surfaces: flat, vertical, and stooped nanohairs (100 nm diameter and 900 nm height). The cells on the flat structures showed random movement while the cells on the vertical hairs showed significantly decreased proliferation. It was also found that the behavior of cells cultured on stooped nanohairs is strongly influenced by the direction of the stooped pattern of hairs when we quantitatively measured the migration of cells on flat, vertical, and stooped structures. Cells on the stooped structure showed higher forward migration preference compared to that of the other structures. Furthermore, we found that these cellular behaviors on the different patterns of nanohairs were affected by intracellular actin flament change. Consistent with these results, the vertical and stooped structures can facilitate the control of cell viability and guide directional migration for biomedical applications such as organogenesis.

Resume : Self-assembled phthalocyanines (Pcs) monolayers have been studied with scanning tunnelling microscopy (STM) due to the organized 2D arrays that they form with potential applications in functional organic devices. The metal central region gives them important electronic, optical and optoelectronic properties and the possibility to extend the 2D monolayer into three dimensions through vertical reaction chemistry [1,2]. Synthetized metal Pcs [3,4] were assembled in graphite and followed by STM. High resolution images were obtained during the formation of the monolayer using a method previously applied in porphyrin monolayers [1,2,5,6]. The molecules were added to graphite trough an interface with tetradecane and the monolayer formation was followed in real time using molecular resolution images obtained by STM. Differential functional theory simulations were made to support the STM results. . [1] Q. Ferreira et al, J. Phys. Chem C, 2014 [2] Q. Ferreira et al, Nanot., 2011 [3] A. Sastre et al, Sust. Energy Fuels, 2017 [4] A. Sastre et al, J. Phys. Chem. C, 2016 [5] Q. Ferreira et al, Applied Surface Science, 273, 220, 2013 [6] Q. Ferreira et al, J. of Microscopy, 2018 Acknowledgements We thank FCT-Portugal, under the project UID/EEA/50008/2013 for financial support

Resume : Dynamic adhesion and detachment of subcellular regions occurs during cell migration. Migrating cells exhibit highly polarized morphology with non-uniform distribution of adhesions across the cells. To systematically investigate roles of adhesions in each region of cells, we devised a new method enabling us to detach defined sub-cellular regions of cells and observe cell responses in real time using a cell-friendly photoresist poly(2,2-dimethoxy nitrobenzyl methacrylate-r-methyl methacrylate-r-poly(ethylene glycol) methacrylate) (PDMP).[1] Unlike conventional photoresist polymers, PDMP dissolves in cell culture medium with near neutral pH conditions after brief UV exposure without harming cells [2], thus in situ micropatterning can be performed in the presence of cells. First, PDMP polymer thin films formed by spincoating on glass coverslips were briefly treated with air plasma and coated with fibronectin. Then, cells were attached on fibronectin-coated PDMP thin films. By using a spatial light modulator, we illuminated UV to specific sub-cellular regions to selectively dissolve PDMP thin films underneath the cells, resulting in detachment of cell adhesions on UV-illuminated regions. Using this new method, we systematically investigated how cells respond when specific sub-cellular regions of the cells were detached.

Resume : Since discovery, organic solar cells (OSCs) have attracted immense research interests because of their unique features such as device structure tunability, lightweight, highly flexible nature, environmental-friendliness. To improve the light harvesting efficiency and photovoltaic performance of OSCs, it is crucial to reduce optical losses including reflections and scatterings. To address this issue, surface texturing, anti-reflection coating and plasmonic nanomaterials have been applied. In this study, we laminated an external M13 bacteriophage textured PDMS film onto OSC device and investigated its efficacy as anti-reflection medium. The OSCs based on PTB7-Th:PC71BM were fabricated on ITO glass substrate. The as-fabricated M13 bacteriophage textured film exhibited high haze and simultaneously, the reflectance of M13 film incorporated OSCs (M13-OSCs) device was lower than that of pristine OSCs. The optical absorption and power conversion efficiency of M13-OSCs was improved. M13 bacteriophage template film could potentially be an excellent metamaterial for future optoelectronics application.

Resume : Silk is a kind of natural macromolecule material with good mechanical properties, and good biocompatibility. The mechanical properties of silk based materials are influenced by hierarchical structure, such as molecular, β-crystalline, and nanofiber structure. Moreover, fibrillar structure, especially at nanoscale, has a crucial effect on the excellent performance of native silk. The properties of silk-based materials are originated from the special arrangement at the mesoscale of hierarchical structures. Conventional ways of preparing silk nanofibrils have many disadvantages, such as, toxic solvent, unstable silk nanofibrils, and low productive rate. And the research of silk nanofibrils is not detailed. To fabricate stable SNFs suspension efficiently, novel solution systems were used. Transmission electron microscope, atomic force microscope, and synchrotron radiation small angle X-ray scattering were applied to confirm the size of SNFs accurately. Molecular dynamics simulations of silk models demonstrated that the potential of mean force required to break the HBs between silk fibroin chains or the Van der Waals’ interactions between β-sheet layers in a particular solvent system. Mesoscale research has an important value on the forming and application of materials and the study of silk nanofibrils has guiding significance to form silk-based materials and construct the hierarchical structure model.

Resume : Poly(3,4-ethylenedioxythiophene) (PEDOT) materials was considered as star materials for fabricating bioelectronic devices due to its excellent electrical conductivity and electrochemical stability. Decoration with functional groups and introducing nanostructures would enhance the electric and biological properties of PEDOT materials, which could expand the potential applications in the fields of bioelectronics. Therefore, it was important to design simple but controllable nano-assembling methods for functionalized PEDOT materials. Compared to traditional template assembling, template-free method had the beneficial of easy fabrication in large scale and integrated preservation of achieved nanostructures, which would make it possible for fabricating nano electronic devices in wafer-scale. Several methods had been reported to realize nano assembling PEDOT materials, however, few of them could realize fabricating different types of nano-morphologies for various functionalized EDOT monomers with one simple method. Herein, we proposed template-free method for fabricating various functionalized PEDOT thin films with nanodot and nanotube morphologies via electrochemical depositing. The morphology parameters could be tuned including diameter, length and density. Glancing incident wide angle XRD was measured to testify the difference of crystal orientation for the achieved nanodot and nanotube thin films. Meanwhile, possible formation reason and process of nanodot and nanotube morphologies were proposed. Besides, simulation tests were carried out and verified the beneficial of the PEDOT and functionalized PEDOT thin films with nanotube morphology using in bioelectronic devices. We hoped the achieved template-free assembling method could be expanded to other kinds of bio-functionalized PEDOT materials and designed PEDOT thin films with proper bio-functional groups and 3D nanomorphologies based on the requirements in practical application of bioelectronic devices.

Resume : Poly(2-methoxyethyl acrylate) (PMEA) shows excellent blood compatibility due to the existence of intermediate water (1). Small amount of amino groups was found to change the hydration structure of 2-hydroxyethyl methacrylate when combining in a copolymer structure, which additionally decreased the interactions with lymphocytes (2). Here we exploit another possibility to manipulate the surface hydration structure of PMEA by incorporation of small amount of other than nitrogen - the hydrophobic fluorine groups in MEA polymers using Atom Transfer Radical Polymerization and the (macro) initiator concept (3). Focusing on the difference in mobility, two kind of fluorinated MEA polymers were synthesized using 2,2,3,3,4,4,5,5,6,6,7,7,8,8-pentadecafluoro-1-octanol (F15) and poly(2,2,2-trifluoroethyl methacrylate) (P3FM) (macro) initiators appearing liquid and solid at room temperature, respectively. The fibrinogen adsorption of the two varieties of fluorinated MEA polymers was different, that could not been explained only by the bulk hydration structure. Contact angle and AFM measurements reveal that the F15-PMEA reorients in water easily to the surface as compared to the P3FM-b-PMEA which reorientation was suppressed by the small solid fluorinated P3FM block. These findings illustrate, that in order to make a better bio-inert material, the chains containing sufficient intermediate water need to be efficiently oriented to the water surface.

Resume : Quartz crystal microbalance (QCM) methods are powerful tool to monitor some phenomena about bio-recognition with real time, which is assembled with flow injection method. QCM-based sensor are simple, convenience, and low cost. In addition, the method can be downsized easily using conventional microfabrication technologies. However, QCM-based sensor has poor sensitivity for low-molecular-weight materials since magnitude of the frequency shift is proportional linearly to the mass change, as a principle of Saurbrey equation [1]. The motivation of the research is improvement of the sensitivity of QCM-based sensor using large surface area due to 3D nanostructures. In this report, an anodic alminum oxide (AAO) was coated on the quartz crystal. The AAO nanostructure was obtained easily by anodizing only aluminum thin film [2]. Then, a self-organized nanohole array having a triangle lattice was formed in the downward direction with a high aspect ratio. The fabricated nanostructure was just like a honeycomb shape. We measured the frequency shift on antigen-antibody interraction of Mouse IgG. The frequency shift on the nano-honeycomb electrode was 2-3 times larger than that on flat Au surface electrode. The limit of detection (LOD) also improved from 0.48 to 0.20 µg/ml. [1] G. Saurbrey, Z. Phys., 155 (1955) 296-222. [2] H. Masuda et al., Adv. Mater., 13 (2001) 189-192.

Resume : Multilayer graphene (MLG) on insulators will lead to various advanced electronic devices. Graphene has a unique two-dimensional structure, whose characteristics are anisotropic. In line with this, large-grained, highly oriented MLG on insulators has been widely investigated. We previously reported the formation of large-grained, highly-oriented MLG on insulators at low temperature by metal-induced layer exchange. However, it was still not clear what type of metal induces the layer exchange growth of MLG. This study investigates the effect of the species of transition metal in the layer exchange growth of MLG. 50-nm-thick metal (Fe, Co, Ni, Ru, Ir, Pt, Ti, Mo, Pd, Cu, Ag, or Au) and 75-nm-thick amorphous carbon (a-C) thin films were prepared onto a glass using magnetron sputtering. The samples were annealed at 600-1000 °C for 1 h. The metal layers were then etched away. Interactions between transition metals and a-C were classified into 4 groups: (1) Layer exchange (Fe, Co, Ni, Ru, Ir, Pt), (2) Carbonization (Ti, Mo), (3) Local formation of MLG (Pd), and (4) No reaction (Cu, Ag, Au). Thus, the layer exchange was achieved for late transition metals. The metals in group (1) were divided into 2 types: low temperature growth or high crystallinity growth of MLG. Pd has an intermediate characteristic because it is located in the middle in the early and late transition metals. The guidelines for selection of catalyst metal species in the layer exchange growth of MLG will be presented based on the periodic table.

Resume : Fe-tannin species have widely been used in our daily life, from food to anti-corrosive products, and recently applied to the interfacial coating with characteristics of non-specific adhesion. Inspired by iron gall ink, a facile strategy was developed for fabricating freestanding films and surfactant-free oil-in-water emulsions through stepwise assembly of Fe(II)-tannin complexes. Fe(II) cations make soluble mono-catecholate complexes with tannins in aqueous solution, and are subsequently oxidized and self-assemble to Fe(III)-tannin complexes for liquid-interface coating. The difference in binding constants of Fe(II)-tannin and Fe(III)-tannin facilitates the oxidation step by lowering the reduction potential of Fe(II). This work provides an insight into the stepwise control of metal-organic complexes for nanobiomedical and nanobiotechnological applications.

Resume : The concentrations of greenhouse gases, especially CO2 and CH4, which play a critical role for global warming and acidification of water sources, reached to new highs in 2015 according to World Meteorological Organization report [1]. There are many reasons why the increasing concentrations of these gases in the atmosphere but the most significant cause is the combustion of fossil fuels in order to supply energy and use as fuel in transportation [2]. With the raising energy demand, the anthropogenic gas emission is increasing every passing day; therefore, the taking under control and storing of these gases have been having a great attention in academic area [3]. Membrane separation, chemical absorption and physical adsorption have been studied and reported as beneficial techniques for capturing of CO2 [4]. Among them, adsorption has been considering one of the most promising technique due to easy usability, low cost and the low energy requirement [5]. Research on the CO2 adsorption on activated carbon (AC) adsorbents has gained significant interest by virtue of their low cost, low regeneration energy, and eco-friendly characteristics [6]. Here in this study, cheap and effective activated carbon was used for CO2 and CH4 gas adsorption. Active carbon with the result thanks to its large surface area it will have both high and high retail attitude, with the adsorption capacities of CO2 and CH4 being very competitive. In summary, we reported AC which are produced from bio based materials can be used for adsorption of CO2 and CH4. Furthermore, a large increase in adsorbing amounts of gases such as CO2 and CH4 has been observed when these carbon materials that are activated have the ability to adsorb ions such as iron. References [1] WMO, The State of Greenhouse Gases in the Atmosphere Based on Global Observations through 2015, World Meteorological Organization, (2016). [2] J.J. He, J.W.F. To, P.C. Psarras, H.P. Yan, T. Atkinson, R.T. Holmes, D. Nordlund, Z.N. Bao, J. Wilcox, 6 (2016). [3] M.G. Plaza, A.S. Gonzalez, F. Rubiera, C. Pevida, J Chem Technol Biot, 90 1592-1600 (2015). [4] J. Fujiki, F.A. Chowdhury, H. Yamada, K. Yogo, Chem Eng J, 307 273-282 (2017). [5] A. Heidari, H. Younesi, A. Rashidi, A.A. Ghoreyshi, Chem Eng J, 254 503-513 (2014). [6] V.K. Gupta, O. Moradi, I. Tyagi, S. Agarwal, H. Sadegh, R. Shahryari-Ghoshekandi, A.S.H. Makhlouf, M. Goodarzi, A. Garshasbi, Crit Rev Env Sci Tec, 46 93-118(2016).

Resume : Additive manufacturing allows for custom-made design of implants and medical devices. Here, the Ti-6Al-4V alloy, which is known to show good biocompatibility, is exploited as printing material of in-vivo used elements. Besides the general materials biocompatibility, the control of cell adhesion on a surface depends strongly on the surface morphology. In this paper, we present an interdisciplinary approach combining the expertise of mechanical engineering and nanotechnology to face this problem on different size scales. On the one hand, we investigate a tailored macroscopic architecture of an additively manufactured element. The freedom of 3D device architecture allows for a local adjustment of the mechanical stability while providing lightweight design. Experimental results and numerical simulations for design rules for an optimum device performance are presented. On the other hand, we show an approach to control the cell-implant interactions on a nanopatterned 3D printed Ti-6Al-4V surface. To this end, we present results on the creation of ordered nanopore arrays by self-assembly processes in block copolymer thin films. The block copolymer lithography allows for large-area surface patterning with sub 20 nm features, which are in particular interesting as the nanopore diameter matches the size of the focal adhesions of a cell allowing for the control of cell adhesion. Both aspects together, we sketch an overall approach for improved implant performance bridging size scales from centimeters to nanometers.

Resume : In the last decade, silica nanoparticles have drawn massive interest by researchers as an excellent carrier numerous viral diseases due its distinguished physical and morphological properties. Higher biocompatibility, large surface to volume ratio, easy surface modifications,high stability and tunable pore sizes along with low cost and easy preparation methods make it more promising towards drug/nucleic acid delivery subsequently followed by larger encapsulation of drug molecules for real-life applications. Gene therapy has become a potential tool in the medical cooperation of genetically caused diseases. DNA or RNA based antiviral strategy showed better potential application over the viral media due to the less chances of gene recombination and immunogenicity. Hence, in this work mesoporous silica nanoparticle (MSN) based carrier system has been synthesized by simple chemical route, for the targeted delivery of DNA molecule against the conserved 5’-untranslated region of a viral RNA molecule to inhibit viral replications as The as-synthesized MSNs have been characterised by Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM), Brunauer–Emmet–Teller (BET) model and Fourier Transform Infrared Spectroscopy (FTIR) studies. The as synthesized MSNs have a diameter in range of 200-300 nm with an average pore size of 8-10 nm and possess very high specific surface area of ~2206 m2/gm. Additionally the synthesized refined MSNs have been conjugated with suitable functional groups to make it a controlled drug delivery system. In vitro cytotoxicity assay in human hepatocyte carcinoma (Huh7) cells exhibits excellent cell viability in presence of these MSNs carriers. Noticeable reducing of viral RNA levels has been achieved in HCV JFH1 infectious cell culture indicating that this nanoparticle based complex molecule can be used as an efficient candidate for the effective delivery of DNA molecule for gene silencing.

Resume : Nowadays the detection of pathogens is an inherent part of environmental or food industry safety. In spite of good selectivity of conventional methods, they are time-consuming and labour intensive. Biosensors are good candidates for real-time monitoring and fast detection of pathogenic agents ‎[1]. Acoustic devices are in the focus of researchers for bio-sensing applications using appropriate surface functionalization ‎[2]. Love wave surface acoustic wave (LW-SAW) sensors possess high sensitivity in liquid ‎[3]. Acoustic energy is confined in the guiding layer close to the sensitive surface, and the energy is not radiated in the liquid due to using pure shear wave with displacement parallel to the surface ‎[4]. Integration of diamond layer brings many favourable properties such as biocompatibility, various biomolecules attachment and prolonged stability of attached biomolecules ‎[5]‎[6]‎[7]. For these reasons, we theoretically investigated properties of layered structures Diamond/SiO2/36°YX LiNbO3 and Diamond/SiO2/ST-cut quartz for potential biosensor applications. Theoretical calculations were carried out with normalized thickness hSiO2/λ in the range of (0.01 – 1) and different thicknesses of the diamond coating. Legendre and Laguerre polynomial approach of wave propagation in layered structures ‎[8] was used for determination of the phase velocity vp and electromechanical coupling coefficient K2 dispersion curves. We also investigated the sensitivity of Diamond/SiO2/ST-cut quartz and Diamond/SiO2/36°YX LiNbO3 structures by using 100 nm thick PMMA film surface loading. The optimal sensitivity of the Diamond/ SiO2/36°YX LiNbO3 is obtained for silicon dioxide normalized thickness hSiO2/λ between 0.3 and 0.6 for all tested diamond thicknesses. In this range of normalized thicknesses hSiO2/λ, the electromechanical coupling coefficient is steeply decreasing from 15 % to 5 %. The diamond/SiO2/ST-cut quartz structure simulation results show similar behaviour. The highest sensitivity and K2 of this structure are obtained for normalized thickness hSiO2/λ between 0.2 and 0.6. Results of this theoretical study show it is possible to fabricate LW-SAW devices with a very thin diamond coating without significant loss of the sensitivity. Properties of LW-SAW devices fabricated on 36° YX LiNbO3 substrate will be presented and compared with theoretical results and previous studies on LW-SAW devices fabricated on ST-cut quartz substrate [9]. [1] SINGH, A. et al., Biosensors and Bioelectronics, 2009, 24(12), 3645-3651. [2] LÄNGE, et al., Analytical and Bioanalytical Chemistry, 2008, 391(5), 1509-1519 [3] GRONEWOLD, Thomas M.A., Analytica Chimica Acta, 2007, 603(2), 119-128. [4] RABUS, D. et al., Journal of Applied Physics, 2015, 118(11), 114505 [5] KRUEGER, Anke a Daniel LANG., Advanced Functional Materials, 22(5), 890-906 [6] Vadym N. Mochalin, et al., Nature Nanotechnology, 7(1):11–23. [7] VAIJAYANTHIMALA, V., et al., Biomaterials, 33(31), 7794-7802 [8] BOU MATAR, Olivier et al., The Journal of the Acoustical Society of America, 2013, 133(3), 1415-1424 [9] Drbohlavová, L. et al., Proceedings 2017, 1, 540.

Resume : Hydrogels are widely used as cell-culture platforms for various biomedical applications. With the biocompatible polymers as building blocks for hydrogels, it is often difficult to provide various physical properties to tailor to specific needs. In this study, bioactive and electrically conductive nanofibers consisting of conductive polymer (PEDOT:PSS) and biopolymer (photocrosslinkable gelatin) are prepared via electrospinning and further processed to generate short, diffusible nanofibers (< 10 um). These short nanofibers are incorporated into gelatin-based hydrogels to improve their mechanical properties as well as electrical conductivity which is otherwise generally non-existent in polymer-based materials. Their properties could be tuned in a wide range by controlling their physical parameters (e.g. concentrations, molar ratio, size, etc.). The nanofiber-infused hydrogel was used as a scaffold for cardiomyocytes to not only improve their viability, but also their electrophysiological functions, which was aided by the presence of conductive nanofibers. Overall, the nanofiber-based composite hydrogel systems presented in this study could provide unique and yet practical 3D cell culture platforms for biomedical applications.

Resume : Bacterial species capable of expressing fluorophores in response to external stimuli are actively being utilized as light-activated sensors for various applications. These stimuli-responsive bacteria encapsulated in miniaturized spherical hydrogels (‘microbeads’) are especially useful as a field deployable form of sensors for detecting environmental chemicals, due to the capability of mass production as well as long-range light detection. Herein, genetically engineered bacteria capable of expressing enhanced green fluorescent protein (eGFP ) in response to nitro compounds were encapsulated into alginate-cellulose beads to develop microbead biosensor. Mechanical strength of the conventional alginate microbeads was improved by incorporating anionic cellulose. The encapsulated bacteria proliferated within the microbeads, and eGFP expression was proportional to the amount of nitro compounds (e.g. DNT and TNT). Furthermore, the M13 bacteriophage having high binding affinity towards DNT and TNT were also encapsulated into the same microbeads, resulting in increased sensitivity of nitro detection. In addition, the fluorescence emitted from the microbead biosensor deployed on a soil sample was detected at long range (e.g. 20 meters and beyond) using a laser fluorescent scanning system to validate the feasibility as field application and safe long-range detection of explosives.

Resume : In this project we studied the mechanical and immunogenic properties of the oxide nanowires hybrid and similar hybrids, to see if in the future they could be used as coatings for medical prosthesis. The first group of samples consisted in nanowires of zinc oxide grown on a thin film of platinum, the second group was characterized by the deposition of graphene oxide on the nanowires. First a metal film of platinum was deposited on glass by RF Magnetron sputtering technique as catalyst. A tube furnace has been used to grow nanowires of ZnO on the top of the catalysed substrate by vapour-liquid-solid (VLS) technique. The samples have been investigated by Scanning Electron Microscope (SEM) to verify the morphology of the nanostructures, confirming the correct growth of the zinc oxide nanowires, then they have been tested for immunogenicity. Peripheral blood mononuclear cells (PBMCs) were isolated from venous peripheral blood of 5 healthy donors. One millions cells were cultured in the presence of zinc oxide nanowires and the same number of cells were cultured with or without stimuli for 5 hours (as positive/negative control of the experiment). In addition, PBMCs with zinc oxide nanowires were cultured for 5 days. No significant signs of lymphocyte activation was detected in terms of cytokine production, either after 5 hours and 5 days of culture.

Resume : Atherosclerosis is a systemic and chronic inflammatory condition in which plaques build up inside the arteries. Accumulation of cholesterol in early lesions leads to the formation of macrophage foam cells that ingest free cholesterol, eventually resulting in the presence of intra- and extracellular cholesterol crystal (CC) in advanced atherosclerotic plaques. Over time, growth of the necrotic core leads to plaque destabilization and vessel narrowing, which in turn increases the risk of rupture and thrombosis, leading to heart attacks and strokes. In this work we have developed novel catalytic and anti-inflammatory polymeric nanomedicines that are capable of directly diminishing a major detrimental effect of atherosclerosis; the formation of CCs within plaques. We present the development and characterisation of targeted polymeric nanobiocatalysts capable of selectively targeting atherosclerotic plaques and ‘dissolving’ CCs via a bioinspired catalytic approach based on innate cholesterol catabolic pathways.

Resume : Atherosclerosis is an inflammation-driven chronic disease of the arteries and the leading cause of death worldwide. Therefore, there is a growing need for efficient drug-testing and drug-screening systems. Commonly used drug-testing technologies are based on two-dimensional cell culture systems, which cannot recapitulate in vivo conditions. On the other hand, animal models are not only lengthy and costly, but also poor predictors of human responses. To overcome these shortcomings, we have proposed to use organ-on-a-chip technology for atherosclerotic nanomedicine studies. We have developed a microfluidic chip consisting of two PDMS layers, separated by a polyester membrane. Each layer has a microfluidic channel, which is capable of simulating the shear condition of a vessel. In addition, endothelial cells were cultured on the membrane and then inflamed to mimic an atherosclerotic vessel. Calcein AM assay was used to investigate cell-viability and morphology of the cells. Moreover, Immunohistochemistry studies and permeation studies were performed. The results of these studies successfully showed tight junctions between cells before inflammation procedure, as well as compromised and leaky junctions after this procedure, which is the main indicator of inflamed vessels. Consequently, the proposed microfluidic chip, which mimics shear condition of a vessel and inflammatory condition of atherosclerosis, is a suitable alternative for typical atherosclerotic drug-screening systems.

Resume : Background: Cardiovascular disease (CVD) is the leading cause of death worldwide – causing 40% of all mortality in Western societies. New therapeutics that can deliver anti-inflammatory biologics in a controlled manner are currently of interest for the treatment of atherosclerosis. As such in this work we have developed matrix metalloproteinase (MMP) sensitive nanogels (NGs) that can release an antioxidant capable of dampening inflammation. MMPs are highly upregulated in atherosclerotic plaques. Results & discussions: For this work a MMP cleavable cross-linker was synthesized by solid phase peptide synthesis methods and characterized by mass spectroscopy, NMR and HPLC. NG polymerization was performed at room temperature using aqueous polymerization techniques. The spherical morphology of the protein loaded NGs was determined using TEM analysis. DLS results were comparable with the TEM size of the protein loaded NGs. Using our methodology high loading and encapsulation efficiencies were obtained due to inclusion of cationic charge of specific acrylate monomers used in the formulations. Conclusions: we have successfully designed and synthesized MMP enzyme cleavable protein loaded NGs systems and optimized the NG synthesis method by changing different parameters.

Resume : magnetic (Fe3O4) nanoparticles coated with platinum i.e. core shell nanoparticles are synthesized through simple and cost-effective co precipitation method. As prepared nanoparticles was studied in detail by using TG-DSC and FTIR for amine coating. TEM images shows formation of spherical core shell nanoparticles with sizes less than 15 nm. High magnetization value of about 59 emu/g at room temperature is obtained for core shell. To control temperature elevation and heat distribution after application of alternation magnetic field and NIR LASER during the internalization of magnetic nanoparticles (MNPs) in magnetic fluid hyperthermia is one of the main challenges. This challenge is well studied in current work.

Resume : The development of new drug delivery (DD) systems able to release the drugs during prolonged periods has been receiving greater attention in recent years due to the ability to use these systems to treat diseases without human intervention. Mainly in ocular diseases with most of treatments consisting in applying eye drops which has a poor patient compliance. We are developing DD multilayers films able to release an ocular drug used in glaucoma treatment which can release the drug during a month and at specific periods of time. Biocompatible films composed of brimonidine encapsulated in β-cyclodextrin alternated with monolayers of a hydrossoluble polymer (poly (β-amino ester)) and/or graphene oxide are able to release a precise amount of drug for a month. The films growth and the pharmacokinetics were monitored by ultraviolet-visible spectroscopy, quartz crystal microbalance and atomic force microscopy. The obtained results showed that the films are stable and drug release can be controlled by the presence of the hydrossoluble polymer and the graphene oxide. In particular, it was observed that graphene oxide delays significantly the brimonidine release enabling precisely control the amount of drug delivered. This work contributed for new developments in DD films that can be used in glaucoma treatment or adapted to other DD systems with other types of drugs. Acknowledgements We thank FCT-Portugal, under the project UID/EEA/50008/2013 for financial support

Resume : The unique physicochemical properties of C60, make it a promising candidate for numerous applications in biomedical, photovoltaic and material science fields. However aggregation phenomena in organic solvents and the lack of solubility in biological environments hamper the exploitation of C60 properties. In this work, different peptidic nanotweezers were designed and synthesized with the aim of dispersing monomolecularly C60 in water. Phenylalanines were used as recognizing moieties, able to interact with C60 through π−π stacking, while a varying number of glycines were used as spacers, to connect the two terminal phenylalanines. The peptidic nanotweezers disperse C60 in water with high efficiency, and the solutions are stable both in pure water and in physiological environments. NMR measurements demonstrated the ability of the peptidic nanotweezers to interact with C60. AFM measurements showed that C60 is monodispersed. Electrospray ionization mass spectrometry determined a stoichiometry of 1C60:4FGGGF. Molecular dynamics simulations, showed that the peptidic nanotweezers assemble around the C60 cage, creating a supramolecular host able to accept C60 in the cavity. Electrochemical and spectroscopic analysis demonstrated that, also upon binding with the oligopeptides, the peculiar properties of C60 were still preserved. The supramolecular complex shows visible light-induced generation of ROS, which make it a suitable sensitizer in photocatalysis or photodynamic therapy.

Resume : The characteristics of a mechanosensor are one of the crucial issues to detect delicate bio-signals for medical applications and fine stress on flexible integrated circuit (IC) electronics. Especially, sensitivity is major issues for the sensor, determining directly the performance of the sensor. Recently, nanocrack based mechanosensor inspired by spider’s vibration receptor has been a breakthrough with its high sensitivity, up to 2,000 in 2% strain, and simple fabrication process. The sensitivity is dramatically enhanced by nano-crack pattern, however, due to nano-cracks, fatigue by repeated stress is concentrated and accumulated on the spot of the crack vertex. Degradation is inevitably compromised even after 1,000 cycle in 2% strain. Thus, to overcome this drawback, we suggest a simple yet robust strategy for remarkable persistence and durability in nanoscale crack based sensor with a self-healable polymer. The self-healable polymer help it make a return to have original shape and performance. Due to the healable property, the sensitivity is stable until 10,000 cycles of 2% strain, and with additional healing at 50 ℃ for 10 minutes, the sensor over 100,000 cycles can be used. External IR LED heating is useful to locally accelerate the healing, not affecting in another component. The proposed strategy can provide high mechanosensitiy as well as highly enhanced durability.

Resume : In this lecture, we will present the recent developments in our group related to circulating tumor cells (CTCs). We will first discuss the utilization of multi-photon microscopy for monitoring CTCs in the blood stream with the help of quantum dots. CTCs are cancer cells that break away from a primary tumor or metastatic site, escape from immunosurveillance, and then circulate in the peripheral blood with the capability of forming distant metastases. The number of CTCs has been used as an indication for the progress of tumor state. However, how CTCs travel in the bloodstream and how they crossed the endothelial barrier are not known. In our group, we have utilized the multi-photon microscopy to study CTCs noninvasively. Pancreatic cancer cells expressing fluorescence were subcutaneously injected to the earlobes of mice forming solid tumor. When the cancer cells break away from the tumor mass, the cancer cells in blood stream can be monitored. The number of CTCs observed in the blood vessels near the tumor mass increased to a maximum value after five week of inoculation. We also tried to identify a sub population of CTCs such as cancer stem cells (CSCs). The trajectories of CTCs and CSCs were measured and analyzed.

Resume : It has been well known to exist DNA and RNA tetraplex structure in a living cell. Teletraplex structures are formed for guanine (G)-rich DNA or RNA sequence with intermolecular or intermolecular folding through G-quartetformation, where four G form plane through Hoogsteen hydrogen bonding formation. Since these tetraplex structures are connected with cancer development, many reserachers have been studying from the standpoint of tetraplex specfic bindder as a new type of anti-cancer drug [1]. We has been focusing naphthalene diimide derivative as tetraplex DNA binder [2]. This derived from the charge transfer type interaction between electron-deficient aromatic plane of NDI and electron-rich aromatic plane of G-quartet. Recently, we found out that cyclic naphthalene diimide (cNDI) shows strong binding to tetraplex DNA with no-binding for duplex DNA[3]. cNDI was synthesized by the connection between both termini of two substituents and this linker part inhibits to bind to DNA duplex with threading intercalation, which is expected to form most stable complex with DNA duplex. On the other hand, cNDI can bind to tetraplex DNA through upper and/or lower G-quartet planes through stacking interaction. cNDI carrying cyclohexane part with linker chain showed the binding affinity with 106 M-1 order for tetraplex DNA, which was over 200-times higher binding affinity for tetraplex DNA than for duplex one using spectra change upon addition of DNAs in the solution of cNDI. The obtained binding affinity using isothermal titration calorimetry (ITC) was in good agreement with that using spectra change for tetraplex DNA, whereas no binding evidence was observed for duplex DNA from ITC. Telomerase repeat amplification protocol (TRAP) assay in the presence of the varied amount of cNDI showed no inhibition for DNA polymerase in PCR and strong inhibition of telomerase activity after elongation until one unit of tetraplex DNA formation. We also synthesized cNDI derivative carring biotin (cNDI-Biotin) to visualize the tetraplex region in a living cell. After added the cNDI-Biotin and Fluorescein Avidin D to the permeabilized interphase U2OS cells, fluorescence spots were observed in nucleolus. This behavior is in agreement with the previous report of tetraplex DNA binder [4]. We need to carry out more detailed experiments, however, this result also show effective anti-cancer drug with low side-effect. References [1] D. Monchaud and M.-P. Teulade-Fichou, Org. Biomol. Chem., 6, 627-636 (2008). [2] S. Sato and S. Takenaka, J. Inorg. Biochem., 167, 21-26 (2017). [3] Y. Esaki, Md. M. Islam, S. Fujii, S. Sato, and S. Takenaka, Chem. Commun., 50: 5967-5969 (2014). [4] D. Drygin, A. Siddiqui-Jain, S. O?Brien, M. Schwaebe, A. Lin, J. Bliesath, C. B. Ho, C. Proffitt, K. Trent, J. P. Whitten, J. K. C. Lim, D. V. Hoff, K. Anderes, and W. G. Rice, Cancer Res, 69, 7653- 7661 (2009).

Resume : Medical decision based on single biomarkers often results in a high possibility of false positives and false negatives. The combination of multiple biomarkers improves accuracy of disease diagnosis. This demonstrates the importance of developing sensitive multiplexing assay of biomarkers to improve reliability of diagnosis. Nano-structure transmission SPR is an emerging technology with potential for multiplex detection and development of POC testing platforms. SPR sensors based on nanostructures facilitate chip-scale integration and allow high-throughput and label-free detections of biomarkers. Nanostructure SPR substrates can be fabricated with inexpensive and high-yield techniques and are therefore suitable candidates for low cost mass production and integration of disposable sensor cartridges. Multiple arrays can be fabricated and by immobilizing different recognition molecule, each array can be used to probe specific target molecule and can be interrogated separately. In this work, we demonstrate a method that employs gold nanoslits (GNS) surface plasmon resonance (SPR) sensor integrated in a microfluidic chip for multiplex detection of miRNA biomarkers. An array of nanoslit structures was designed and fabricated on an olefin polymer film using nanoimprint lithography and 80 nm gold was deposited. The approach for detection include double hybridizations by the capturing probe I on the Gold nanoparticles (AuNPs) and the second capturing probe II immobilized on the GNS array. The double hybridization with two probes complementary to the target molecules results in high specificity of detection. AuNPs used for the signal enhancement of SPR. In this work, target miRNA as low as 100 fM were detected simultaneously without any labeling and amplification. The high sensitivity of this method is attributed to the AuNPs-enhanced transmission SPR on the gold nanoslits through increased surface mass, high dielectric constant of Au particles and electromagnetic coupling between AuNPs and the extraordinary optical SPR transmission of gold nanoslits.

Resume : Live virus vaccines elicit profound T-cell and B-cell memory responses for effective antiviral immunity, yet these viral vectors pose safety concerns and are difficult to develop for pathogens of high biosafety levels. To accelerate vaccine development, nanotechnology has been applied to enhance the potency of subunit vaccines. Inspired by the immune-potentiating effect of virions, we engineered a synthetic virus-mimetic nanoparticle vaccine with potent and tunable immune potentiating capability by incorporating advanced nanotechnology with recombinant protein techniques. The nanoparticles are designed with multiple virus-like features, including (i) a synthetic polymeric capsid consisting of a biocompatible and biodegradable polymer with controllable size and stability, (ii) an aqueous inner core readily loaded with adjuvants for immune activation, and (iii) surface functionalization with virus subunit proteins for antigen presentation. Encapsulation of a STING agonist inside the polymeric capsid significantly improves the adjuvant?s immune-potentiating effect and safety, elevating cytokine production in the lymph node and yet reducing serum cytokine levels following subcutaneous administration. In light of the ongoing outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV), which has caused 2189 cases and 782 deaths in 27 countries since 2012, a MERS-CoV vaccine was prepared based on the nanoparticle technology. Using a highly conserved receptor binding domain (RBD) of MERS-CoV spike protein, the nanoparticle vaccine demonstrates extraordinary capacity in promoting humoral and cellular responses, inducing elevated levels of antigen-specific antibodies, CD4+ T lymphocytes, and CD8+ T lymphocytes. In addition, an increased number of CD44+CD62L+ memory T cells is also observed. In a mouse model of MERS-CoV infection, the nanoparticle vaccine potently inhibits virus proliferation and protects the mice from virus-induced mortality. The nanoparticle is readily amenable to other pathogenic targets, offering a versatile platform for vaccine design.

Resume : Nerve cell culture takes irreplaceable roles in molecular and cellular neuroscience. However, its use in systems-level studies has been hindered due to the substantial difference in neuronal connectivity from the actual brain. Here we employed surface engineering technology to prepare guidance cues to extrinsically direct the development of cultured neurons and investigated how the modulation of network structure at the mesoscopic scale influences the network dynamics. We focused on the modular organization of brain networks, characterized by the presence of densely-connected subsystems, i.e., modules, that are weakly interacting with each other. Fluorescence calcium imaging of spontaneous neural activity shows that the atypical dynamics of the cultured networks, characterized by a bursting activity that is highly-synchronized across the whole network, can be suppressed by the induction of modular organization in the networks. Increasing the degree of modularization caused the networks to generate activity patterns that were spatially and temporally more complex. Our results demonstrate that surface micropatterning provides a unique tool to constructively study the structure-function relationships in living neuronal networks.

Resume : Silica is generally recognized as safe (GRAS) and recently a kind of silica nanoparticles known as Cornell dots, or C dots, have received an investigational new drug (IND) approval for the first-in-human clinical trial for targeted molecular imaging of cancer by FDA. Mesoporous silica nanoparticles (MSN) are intriguing nanocarriers for efficient and cell-specific delivery of proteins, enzymes, and anti-cancer drugs to improve treatment of diseases. However, despite often highly promising in vitro findings, such as enhanced uptake and intracellular processing as well as efficacy, practical applications of MSN are usually limited due to poor stability, serious aggregation and short in vivo circulation lifetimes in biological media. The previous study has demonstrated that the nanoparticle flow, margination and adhesive properties in blood vessels are dependent on particle size, surface charge, hydrophobicity, and geometry. For cancer drug delivery, PEGylation of nanotherapeutics is usually applied to have a prolonged blood circulation half-life, enhancing their possibility to extravasate through the leaky tumor vasculature. Here we synthesized a serious of PEGylated MSNs and systematically examined the effect of size, charge and especially surface heterogeneity (i.e., spatial arrangement and relative exposure of chemical motifs) of PEGylated MSNs on their biological fate. In vivo study have been dedicated to quantitatively as well as qualitatively investigate the time evolution of protein corona formed on MSN, and understand how protein corona mediates the tumor accumulation as well as urinary excretion of nanocarriers.

Resume : Transcription factor complex NF-kB (p65/p50) is localized to the cytoplasm by its inhibitor IkB-alpha. Upon activation, the Rel proteins p65/p50 are released from IkB-alpha and transported through nuclear pore to effect many gene expressions. While inhibitions of up or down stream signal pathway are often ineffective due to cross talks and compensations, direct blocking of the Rel proteins p65/p50 has long been proposed as a potential target for cancer therapy. In this work, a nanoparticle/antibody complex targeting NF-kB is employed to catch the Rel protein p65 in perinuclear region and thus blocking the translocation near the nuclear pore gate. TAT peptide conjugated on mesoporous silica nanoparticles (MSN) help non-endocytosis cell-membrane transducing and converge toward perinuclear region, where the p65 specific antibody performed the targeting and catching against active NF-kB p65 effectively. The size of the p65 bound nanoparticle becomes too big to enter nucleus. The new approach of antibody therapy targeting on transcription factor with ?nucleus focusing? and ?size exclusion blocking? effects of the antibody-conjugated nanoparticle is general and may be applicable to modulating other transcription factors.

Resume : A robust nanopillar platform with increased spatial resolution reveals that perinuclear forces originated from stress-fibers spanning the nucleus of fibroblasts are significantly higher on these nanostructured substrates than the forces acting on peripheral adhesions. Many perinuclear adhesions embrace several nanopillars at once, pulling them into ?1-integrin and zyxin rich clusters, which are able to translocate in the direction of cell motion without loosing their tensile strength. The high perinuclear forces are greatly reduced upon inhibition of cell contractility or actin polymerization, and disruption of the actin-cap by KASH dominant-negative mutant expression. LMNA null fibroblasts have higher peripheral versus perinuclear forces, impaired perinuclear ?1-integrin recruitment as well as Yap nuclear translocation, functional alterations that can be rescued by lamin A expression. These highly-tensed actin-cap fibers are required for YAP nuclear signaling and thus playing far more important roles in sensing nanotopographies and mechano-chemical signal conversion than previously thought.

Resume : The start of my research was critical to the advance of cryo-electron microscopy, allowing researchers to obtain images of biological materials that more closely resembled the natural state of the material. I continued to refine techniques for structural imaging of biological materials by cryo-electron microscopy and developed a method known as cryoEM of vitreous sections (CEMOVIS), which researchers could apply to the vitrification of cells and tissues for the visualization of very fine structural detail. Also I continued to apply electron microscopy to the study of structural aspects of DNA and chromatin. The basic results obtained on functional transitions of DNA structure. DNA molecules change their structure during such vital processes like DNA replication, DNA recombination, DNA repair or DNA transcription. Understanding these functional transitions of DNA structure helps to elucidate molecular mechanisms involved in normal and pathological situations like for example in patients with defective DNA repair. Long term aim of these investigations is to catalogue various functional DNA structures and to understand the effect of such factors like nucleotide sequence, DNA supercoiling, counter ion concentration, mutagens and therapeutic drugs on affecting these functional transitions of DNA structure. With cryo-EM researchers can advance understanding of Life’s chemistry and develop pharmaceuticals. A selection Jacques Dubochet’s “Nobel Scientific Publications” are now Open Access in UNIL’s institutional repository 1. Cryo-electron microscopy of vitrified specimens Dubochet J., Adrian M., Chang J. J., Homo J. C., Lepault J., McDowall A. W., Schultz P., 1988. Quarterly Reviews of Biophysics, 21 (2) pp. 129-228. 2. Emerging techniques: Cryo-electron microscopy of vitrified biological specimens Dubochet J., Adrian M., Lepault J., McDowall A.W., 1985. Trends in Biochemical Sciences, 10 (4) pp. 143-146. 3. Geometry and physics of catenanes applied to the study of DNA replication Laurie B., Katritch V., Sogo J., Koller T., Dubochet J., Stasiak A., 1998. Biophysical Journal, 74 (6) pp. 2815-2822. 4. High-pressure freezing for cryoelectron microscopy Dubochet J., 1995. Trends in Cell Biology, 5 (9) pp. 366-8. 5. Brownian dynamics simulation of DNA condensation Sottas P.E., Larquet E., Stasiak A., Dubochet J., 1999. Biophysical Journal, 77 (4) pp. 1858-1870. 6. Twisting in a crowd Dubochet J., 1993. Trends in Cell Biology, 3 (1) pp. 1-3. 7. A Reminiscence about Early Times of Vitreous Water in Electron Cryomicroscopy, Dubochet J., 2016. Biophysical Journal, 110 (4) pp. 756-757.

Resume : The integration of carbon nanoparticles (CNPs) with proteins to form hybrid functional assemblies is an innovative research area with great promise for medicine, nanotechnology, and materials science [1,2]. The chemical and physical properties of CNPs and proteins differ greatly, but the ?size-commensurability? makes possible their interaction [1,2]. Using C60 [1,3-5] and CNT (10,10) as CNPs models and adapting protocols widely used in drug design (virtual screening and reverse docking), the more probable CNP binding proteins were identified. A computational ?baiting protocol? fish out from a database the more suited proteins able to recognize the selected CNP. The recognition process is highly specific and localized to a well-defined protein pocket [3-5]. The effect of the binding, at the level of the single amino acid, is characterized by a variety of experimental and computational approaches [3-5]. [1] M. Calvaresi, F. Zerbetto, Acc. Chem. Res. 2013, 46, 2454. [2] M. Calvaresi, F. Zerbetto, ACS Nano 2010, 4, 2283. [3] M. Calvaresi et al. ACS Nano, 2014, 8, 1871. [4] M. Calvaresi, A. Bottoni, F. Zerbetto, J. Phys. Chem. C 2015, 119, 28077. [5] M. Di Giosia, M. Calvaresi et al. Nanoscale, 2018, DOI:10.1039/C8NR02220H

Resume : The rapid, accurate, non-expensive and wireless measured detection of low concentrations of biological or chemical compounds is, currently, crucial in the combat of serious public health concern. Magnetoelastic resonant platforms of metallic glass ribbons are being widely spread for sensing applications due to their change on resonant frequency in response to differences in physical parameters such as stress, pressure or mass loading. The performance of theses sensors has been object of study of many investigations, however, most of them are based on an increase of sensitivity by a reduction of the magnetoelastic ribbon size. Despite the interesting obtained results, it forces to work on high frequency values and generates lower magnetic field, requiring a better detector. In order to avoid the reduction of magnetoelastic ribbon size, this research propose an approach based on the variation of the magnetoelastic ribbon shape and on the percentage of ribbon?s area covered by the specific target. The traditional rectangular shape of magnetoelastic ribbons were replaced by triangular shape with different length and widths. Home made Fe64Co21B15 metallic glass were used in this work. The present work show that mataining the size of the resonator, the change of its shape result in an increase of more than 400% of its sensitivity.

Resume : Keynote Forum E- MRS Symposium E: Scanning tunnelling microscopy (STM) is the elected technique by the scientists to visualize and manipulate matter at molecular/atomic scale. Namely, has been used to monitor in real-time the formation of self-assembled monolayers highly organized in which the molecules arrange themselves into packed 2D crystals and fully covering the surface[1,2]. Several high resolution STM images about self-assembly molecular systems formation have been reported revealing the parameters which are involved in the monolayers formation whether they are physical parameters (e.g. concentration of solutes, wettability, contact angles, superficial tension, temperature variations) or chemical parameters (e.g. molecule structure, orbital configuration). Recently, the STM has been used to build molecular systems with multicomponents, e.g., self-assembled monolayers with more than one molecular element, vertical supramolecular structures synthetized in-situ[3,4] and/or molecular switches[2]. This work reviews the methodology which is involved in the development of these systems revealing details on how to use the STM to monitor their fabrication[5]. References: [1] Q. Ferreira, A. M. Bragança, N. M. M. Moura, M. A. F. Faustino, L. Alcácer, J. Morgado, ?Dynamics of porphyrin adsorption on highly oriented pyrolytic graphite monitored by scanning tunnelling microscopy at the liquid/solid interface?, Applied Surface Science, 273, 220, 2013. [2] J. Oliveira, A. M. Bragança, L. Alcácer, J. Morgado, Mário A. T. Figueiredo, J. Bioucas, Q. Ferreira, Sparse-coding denoising applied to reversible conformational switching of a porphyrin self-assembled monolayer induced by scanning tunnelling microscopy: IMAGE DENOISING ALGORITHMS FOR STM IMAGES, Journal of Microscopy, DOI: 10.1111/jmi.12699, 2018 [3] Q. Ferreira, A. M. Bragança, L. Alcácer, J. Morgado, ?Conductance of well-defined porphyrin self-assembled molecular wires up to 14 nm in length?, Journal of Physical Chemistry C, 118 (3), 7229 - 7234, 2014. [4] Q. Ferreira, L. Alcácer, J. Morgado, ?Stepwise Preparation and Characterization of Molecular Wires made of Zinc octaethylporphyrin complexes bridged by 4,4?-bipyridine on HOPG?, Nanotechnology, 22, 435604, 2011 [5] C. Delfino, Q. Ferreira, Recent Advances in bottom-up self-assembled supramolecular structures builded by STM, Materials, 2018, accepted. Acknowledgements We thank FCT-Portugal, under the project UID/EEA/50008/2013 for financial support

Resume : Physicochemical methods of testing of innovative technologies products are widely used in laboratory, manufacturing or normative practices. Those methods allow to determine the expected quality of the product and its conformity with applicable norms increasing the safety of the product use. Products for biomedical applications require further biological evaluation, as presented in a number of legal norms. In connection with the above popular is a position, additionally supported by norms, that only industrial products for medical applications must be subjected to biological evaluation. This standpoint is often presented dogmatically by representatives of technical sciences, indisputably denying the need to use biological methods in material research, with the obvious exclusion of biomaterials. As an argument, the statement is usually made that the product is not intended to come into contact with living organisms, and therefore its biological assessment makes no sense. What's more, it is argued that since for hundreds of years of industrial production of various goods, lack of biological evaluation was no problem, nowadays such a problem is not present or is unnecessarily raised. Meanwhile, the problem exists and seems to be quite serious. First of all, attention should be paid to the rate of development of modern science and the accompanying rate of emergence of new technologies, and thus also the rate of appearance of new products, so far not present in our environment. Secondly, special attention should be paid to the presence and intensity of degradation processes of products, including microbial degradation. Slightly simplifying, it can be stated that in the past living organisms had enough time to start and consolidate adaptation processes in response to new industrial products emerging in the environment, currently treated as traditional, as well as products of their degradation. It is worth noting that products of materials degradation can easily get to the biosphere through microorganisms and plants - and thus they will appear in the food chain, at the end of which we are. Of course, the safety of use of innovative materials is not the only benefit of biological evaluation. No less important are new data allowing better designing and selection of appropriate materials for implants manufacturing, production of newest diagnostic systems as well as introduction of intelligent and well controlled drug delivery systems in personalized medicine. Integration of all data concerning processes, structures and properties of materials collected at all relevant scales, from nano to macro, using of all available physicochemical and biological methods, is currently in the focus of modern materials engineering and is called as MATERIOMICS. This term has been proposed by Akita et al. (AIST Japan) in 2004 and is currently creatively developed by Markus J. Buehler (MIT/USA), Clemens van Blitterswijk, Jan de Boer and H. Unadkat (University of Twente/The Netherlands) and others. Integration of physicochemical data, including quantum physics data, with cellular response to exposure to the material/nanomaterial, observed at molecular level, gives us possibility to identify this material/nanomaterial in the case the classical physicochemical methods are not sufficient. Our combined team of the Biophysics Department of Lodz University of Technology and GLP certified laboratories of Bionanopark Ltd has access to and uses physicochemical and biological methods to comprehensive evaluation of materials/nanomaterials and their degradation products. Selected test results will be presented during the lecture. Acknowledgement: The work was supported by the Polish National Center for Research and Development project POIR.04.01.04-00-0058/17-00.

Resume : In the last five years, a lot of research effort has been devoted to the creation of hybrid materials which change the electronic properties of one constituent by changing the optoelectronic properties of the other one. The most appealing approach consists on the interaction between organic materials or metals with biological system such as proteins or DNA. Although experimental efforts have already resulted in the formation of a number of stable hybrid bio-organic materials, the main bottleneck of this research field is the formation of the interface between the biological part and the organic/metal one. In particular, the efficiency of the final devices is very low due to problems with the interfacing of such different materials, charge recombination at the interface and the high possibility of losing the function of the biological component which leads to inactivation of the device. Here, we present a multiscale computational design which allow the study of complex interfaces for stable and highly efficient hybrid materials for biomimetic application. In particular, we focus on the use of graphene as organic material/metal and light harvesting protein complexes (Photosystem I) as biological counterpart, linked together via a self-assembly monolayer (SAM) and a biological linker (cytochrome C) to allow flexibility of the whole system, in order to create novel biomimetic materials for solar-to-fuel, bio-transistors or bioorganic electronic applications.

Resume : Molecularly imprinted polymers (MIPs) are excellent example of bio-mimicking recognition materials. They have found numerous applications in selective chemosensing. For electrochemical determination of electroinactive analytes, usually some external redox probe is added. It is assumed that binding of target analyte molecules by MIP molecular cavities causes MIP film swelling or shrinking. This behavior leads to changes in MIP film permittivity for the redox probe and thus changes in faradaic currents corresponding to reduction or oxidation of the probe (so called ?gating effect?) in CV and DPV determinations. However, this mechanism seems to be inadequate for electrochemical sensors with conductive MIP film recognition units. It was proven that drop of the redox probe oxidation peak in DPV determination originates from changes in electrochemical properties of the MIP film. We can speculate that diffusion of a redox probe is a not crucial issue in terms of selective determination with the MIP film coated electrode. Therefore, a new specially designed monomer, vis., p-bis(2,2'-bithien-5-yl)methyl-ferrocene benzene was used for deposition of a self-reporting MIP film. This monomer acted as both a crosslinking monomer and an internal redox probe. The self-reporting MIP film modified electrodes were used for electrochemical determination of p-synephrine ? a diet supplement that is suspected of causing serious cardiovascular diseases - in the absence of the external redox probe.

Resume : Cells in our bodies are arranged in three dimensions (3D) according to complex architectures, which play a fundamental role in the functionality of the living tissue. There is therefore an urgent need to develop 3D architectures featuring biomaterials that provide the biocompatibility and the physico-chemical properties necessary for the proliferation, differentiation and functionality of living cells. Here, we report the realization of PEGDA biocompatible 3D hydrogel architectures for supporting neuronal cell growth [1,2]. Thanks to the porous nature of the proposed scaffold geometry, realized by two-photon direct laser writing (2P-DLW) [3], we observed the efficient growth of a ramified neuronal network featuring multiple neuritic extensions per cell. Scanning electron microscopy characterization has been associated to two-photon confocal imaging in order to shed light on the localization and the morphology of cells not only around the 3D scaffold but also within its most inaccessible core regions. PEGDA hydrogel showed very low intrinsic fluorescent emission (?100 times lower than conventional polymeric materials employed in 2P-DLW), thus enabling a multi-staining immunofluorescence evaluation of the functional features of neuro2A cells in three dimensions. These unique ?quasi-transparency? optical properties allowed a full 3D immunofluorescence reconstruction of the cell colonization as well as the detection of F-Actin microfilaments and ?-tubulin neuronal marker even ?through? the PEGDA biomaterial. [1] A. Accardo et al., Mater. Today 2018, 21 (3), 315-316. (Front Cover) [2] A. Accardo et al., Biomed. Phys. Eng. Express 2018, 4, 027009. [3] A. Accardo et al., Small 2017, 13 (27), 1700621. (Back Cover)

Resume : INTRODUCTION Ischemic stroke occurs due to reduced perfusion to a brain region, because of the blockage of a blood vessel, resulting in death or permanent neurological deficits. Although, many treatments that make use of therapeutic nanoparticles have been presented, unfortunately to date, no effective treatment has been found to prevent damage to the ischemic brain after stroke. Oxidative stress is related to the pathogenesis of stroke, and overproduction of reactive oxygen (ROS) and reactive nitrogen species (RNS) are thought to be the main cause of this. In view of this, we hypothesized that the targeted delivery of antioxidant (CeO2) nanoparticles (NPs) and iNOS inhibitors (i.e. L-NIL) across the blood brain barrier (BBB) will reduce the overproduced ROS and RNS, resulting to the amelioration of the neurological deficits caused by oxidative stress. METHODS The fabrication of the biomimetic and neuroprotective nanocapsules (BIONICS) was achieved using a hot-emulsion ultra-sonication method, while their characterization was achieved using various material characterization techniques (SEM, TEM, EDX, ICP, FT-IR, TGA). The stability of the nanoparticles was studied using DLS in various biologically relevant media and under conditions that simulate oxidative stress at different time points. The antioxidant ability of CeO2 NPs was studied using EPR as well as antioxidant assay kits. A nitric oxide assay kit was also used to assess the ability of CeO2 NPs and L-NIL to inhibit the overproduction of RNS. The internalization of the LMNVs was studied at various time points using confocal microscopy and their ability to inhibit oxidative stress in vitro was tested on human astrocytes and endothelial cells using flow cytometry. RESULTS AND DISCUSSION The synthesized nanosystem that had an average size of 150 nm, comprised of 70% of a lipid matrix and 30% of CeO2 NPs. BIONICS presented excellent short-term and long-term stability in aqueous media, while it?s stability was affected under conditions simulating oxidative stress. EPR results showed that the CeO2 NP-loaded nanocapsules demonstrated excellent antioxidant capacity which was similar to the antioxidant capacity of the corresponding plain CeO2 NPs. In addition, CeO2 NPs and L-NIL were found able to scavenge overexpressed RNS. Confocal imaging demonstrated a time-dependent internalization of BIONICS while flow cytometry results demonstrated that BIONICS has a protective effect against oxidative stress by inhibiting ROS-mediated apoptosis. CONCLUSION BIONICS demonstrated excellent short-term and long-term stability as well as excellent antioxidant capacity, resulting in the inhibition of ROS and RNS mediated oxidative stress, and in a subsequent neuroprotection of astrocytes and endothelial cells. ACKNOWLEDGMENTS ?This project has received funding from the European Union?s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 793644?.

Resume : INTRODUCTION Glioblastoma multiforme (GBM) is considered one of the most aggressive malignancies in the brain due to its invasiveness and rapid growth, resulting in more than 90% 5-year mortality [1]. Current treatment strategies include tumor resection using combinatory radiotherapy and chemotherapy, but fail to successfully treat GBM due to various reasons, including the location and the pathophysiology of the tumor and the blood brain barrier (BBB) that hinders an efficient delivery of various anticancer drugs [2]. In view of this, we hypothesized that the fabrication of a biomimetic drug delivery system able to cross the BBB and to controllably deliver the anticancer drug temozolomide (TMZ), as well as the simultaneous magnetic hyperthermia treatment, will result into a tumor regression increasing the life expectancy of patients suffering from GBM. The encapsulated SPIONs inside the lipid matrix will also allow the fabricated system to be used as a versatile theranostic. METHODS The fabrication of the lipid-based magnetic nanovectors (LMNVs) was achieved using a combination of hot ultra-sonication and high-pressure homogenization (HPH), while their characterization was achieved using various material characterization techniques (TEM, EDX, FT-IR, TGA, SQUID). DLS was used to assess the stability of the LMNVs at various temperatures and also the formation of protein corona at various time points. Loading and release studies were performed using high-pressure liquid chromatography and the magnetic hyperthermia effect was demonstrated using an alternating magnetic field (AMF). NMR relaxometry measurements were used to assess the ability of the LMNVs to be used as theranostic vectors. The internalization of the LMNVs was studied at various time points using confocal microscopy and their apoptotic effect was assessed using flow cytometry. An in vitro BBB model was used to assess the targeting ability of peptide-functionalized LMNVs. RESULTS AND DISCUSSION LMNVs of 100 nm were successfully synthesized and characterized. The LMNVs demonstrated high stability at various temperatures, and after the protein corona formation. The ability of the LMNVs to increase temperature in a controlled manner at temperatures above 42°C (mild hyperthermia) was assessed by using an AMF, and the results demonstrated that LMNVs need 8.5 min to increase the temperature from 37°C to 42°C. LMNVs demonstrated a good loading capacity of TMZ and a sustained released profile. Confocal microscopy images demonstrated a time-dependent internalization of the LMNVs while viability studies on U87 cells demonstrated that the LMNVs are not toxic (400 ?g/cm2, 72h). In vitro studies under an AMF demonstrated a controlled increase in apoptosis (8% - 30%) depending the time of exposure, in comparison with cells under an AMF without LMNVs treatment. Preliminary results showed that functionalized LMNVs can penetrate the in vitro BBB model. CONCLUSION The LMNVs present high stability in biologically relevant fluids and a good drug release profile. These vectors have the ability to induce mild hyperthermia at a short time (8.5 min) and demonstrate an enhanced apoptotic effect against U87 cells depending on the time of exposure under the AMF. The peptide-functionalized LMNVs can cross the in vitro model of BBB and can be potentially used as theranostic vectors. ACKNOWLEDGMENTS This project has received funding from the European Research Council (ERC) under the European Union?s Horizon 2020 research and innovation program (grant agreement N°709613, SLaMM). REFERENCES [1] S. K. Carlsson et al., EMBO Mol Med 2014, 6, 1359 [2] a) A. Bhowmik, et al., Biomed Res Int 2015, 2015, 320941; b) S. K. Carlsson et al., EMBO Mol Med 2014, 6, 1359; c) G. Iacob & E. B. Dinca, J Med Life 2009, 2, 386.

Resume : INTRODUCTION Glioblastoma multiforme (GBM) is a malignant tumor of the central nervous system (CNS). Nearly 90% of the CNS astrocytic tumors are classified as GBM and are responsible for the extremely low survival rates worldwide. Nowadays, the treatment for GBM consists of a surgical resection followed by an adjuvant chemotherapy, radiotherapy and/or magnetic hyperthermia, methods which are still inadequate to fully cure this disease when used by themselves. Smart drug delivery systems (SDDS) represent another way to treat GBM by holding attractive characteristics like biomimicry, easiness in crossing the blood brain barrier (BBB) and multi-stimuli responsiveness, that enhance the therapeutic effectiveness and reduce the side-effects of the current treatment strategies. In this study, we hypothesized that the coating, with a GBM cell membrane, of a composite nanoparticle consisting of a magnetic core and an antioxidant shell (MANP), and its loading with the drug temozolomide (TMZ), could represent a novel system with an inherent targeting ability that can effectively treat GBM. These MANPs have also the potential to be used for guided delivery and as magnetic resonance imaging contrast agents. METHODS The MANPs were prepared using a simple one-pot hydrothermal method, while their coating with cell membrane was achieved using high-pressure homogenization (HPH) and ultrasonication. The composite system was fully characterized using a variety of techniques including, SEM, EDX, FT-IR, TGA, SQUID, XRD, XPS, BET and ICP. The hydrodynamic diameter and the surface charge of the MANPs as well as the protein corona formation, was studied in media with different ionic strengths (water, DMEM and DMEM with 10% of FBS) at different temperatures and time points, using DLS. An alternating magnetic field (AMF) was used to assess the ability of the MANPs to increase temperature above 42°C while an oxygen sensor was used to assess their ability for ROS-mediated oxygen generation. NMR relaxometric studies were used to assess the ability of the MANPs to act as contrast agents. Release studies were performed using high-pressure liquid chromatography (HPLC) while antioxidant analysis were carried out using various assay kits. Confocal microscopy was used to check the internalization of the MANPs, while preliminary viability tests were fulfilled using Alamar Blue. The apoptotic effect was studied using flow cytometry. RESULTS AND DISCUSSION Electron microscopy showed that the MANPs have a diameter of about 40nm and are stable in water and in DMEM with 10% FBS, in contrast to plain DMEM. Protein corona formation is achieved in the first few minutes after the MANPs interact with serum albumin, and this protein corona is responsible for the increased stability of the MANPs in high ionic strength media. MANPs have the ability to generate oxygen after reacting with H2O2, while their exposure to an alternating magnetic field showed that a temperature useful for hyperthermia treatment (> 42°C) is reachable in almost 5 minutes. Confocal microscopy studies showed a better internalization of cell membrane-coated NPs in U251 cells with respect to the internalization of lipid coated NPs (cetyl palmitate/glyceryl monooleate/mPEG-DSPE) that were used as control. Finally, the combination of hyperthermia and TMZ was more effective on killing GBM cells compared to hyperthermia or TMZ alone. CONCLUSION The MANPs can be potentially used as theranostic vectors. Indeed, due to their magnetic core they can be used for imaging, guided delivery and controlled release, while their antioxidant shell act as a ROS-scavenger and oxygen generator that enhances the efficacy of chemotherapeutic agents like TMZ. The cell membrane coating enhances their biocompatibility as well as their uptake by GBM cells demonstrating an inherent targeting ability without further functionalization. ACKNOWLEDGMENTS This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement N°709613, SLaMM).

Resume : Bones recovered from archaeological excavations are not simple objects. They represent a true “Biological Archive” that deserves to be available and accessible for long-term studies. Materials that have been used for the consolidation of bone since the start of the twentieth century mainly include natural and synthetic adhesives (vinyl and acrylic polymers and copolymers, alkoxysilanes) [1]. Unfortunately, these materials cause many problems due to their scarce chemical stability (the natural ageing of these conservation products rapidly leads to their yellowing over time, formation of crosslinking that result in reduced solubility and irreversibility of the treatment) and to their poor penetration, embrittlement, shrinkage resulting in delamination and formation of local detachments. Moreover, in addition to possible damage they can also limit the possible re-treatability of the object. The development of more proper and affordable strategies for the restoration and conservation of ancient bones is therefore crucial to maintaining long-term access to the biological information recorded in the bones. In order to increase both the durability of the treatment and the physio-chemical compatibility between the matrix and the consolidating agent, a possible approach is based on the use of consolidants constituted by inorganic materials. The main goal of this work is the long-term preservation of bone finds of historical and archaeological interest that has been achieved through the development of innovative smart nanostructured materials with enhanced both physical and chemical properties. A key requirement is that the developed materials must be sustainable and totally chemically compatible with the porous support to be preserved/restored. In order to improve the chemical compatibility with the matrix of the bone respect to the procedures commonly used for the consolidation of bones, the attention has been focused on the possibility to use hydroxyapatite (HAP) as consolidating agent. Two different strategies have been followed: first the consolidation has been carried out through the application of HAP nanoparticles (HAP NPs), then we also tried to induce the crystallization of HAP directly into the porous matrix of the bone. Furthermore, in a previous paper [2, 3] it has been demonstrated that the chemical composition of a bone can favour the formation in-situ of aragonite, through the carbonation of Ca(OH)2 nanoparticles. This mineral, due to its high hardness (2.25 GPa vs 1.54 GPa of calcite) should favour a strong increase of the hardness of the consolidated relic (up to 200%). Then, the action of the HAP has been coordinated with the one of Ca(OH)2 nanoparticles in order to maximize at the same time both the improvement of the mechanical properties and the compatibility of the consolidation treatment. The treated bone relics have been characterized in terms of overall porosity and surface area through gas porosimetry technique: whilst no substantially changes has been observed after application of only NPs, the in-situ precipitation of hydroxyapatite with or without pre-treatment with Ca(OH)2 NPs affects the overall porosity and the BET adsorption-desorption isotherms indicate a decrease of surface area and pore volume of 45% and 64% respectively. Furthermore, X-ray microtomography images clearly show that the less dense regions decrease after treatment instead of a homogeneous denser phase, successfully confirm the consolidating effect on the bone relics. The innovative method for the consolidation of bone relics presented in this work and based on the use of a nanostructured inorganic material (a blend of HAP and CA(OH)2 NPs), represents a promising, durable and performing alternative to the traditional polymeric consolidants, commonly used at today for the treatment of bone remains. References [1] Johnson, J. J. (1994) Consolidation of Archaeological Bone: A Conservation Perspective, Studies in Conservation 21, 221–233. [2] Natali I., Tempesti P., Carretti E., Potenza M., Sansoni S., Baglioni P., Dei L. (2014) Aragonite Crystals Grown on Bones by Reaction of CO2 with Nanostructured Ca(OH)2 in the Presence of Collagen. Implications in Archaeology and Paleontology, Langmuir 30, 660-668. [3] Baglioni P., Carretti E., Chelazzi D. (2015) Nanomaterials in art conservation, Nature Nanotechnology 10, 287-290.

Resume : Drug delivery systems are divided into two main categories: passive systems and active ones. The active systems involve remote controlled drug delivery chips, which releases a certain dose of drug on demand from outside the body. Both systems are designed to facilitate cancer treatment and prevents the patient to get involved with the chemotherapy's side effects. In the present study, the novel implantable drug delivery chip was designed by silicon reservoir and Ionic Polymer Metal Composite (IPMC) actuator integration. IPMC was fabricated via chemical electroless plating technique that resulted in high quality electrode deposition with remarkable IPMC performance. IPMC actuator and silicon reservoir were coupled together and Polydimethylsiloxane (PDMS) polymer was used as the sealing sheet on the reservoir gate. The reservoir was developed by photolithography and etching and next the IPMC strip was attached as the gate of the drug reservoir. The whole design was tested to be biocompatible and its biocompatibility was tested by the MTT assay. The IPMC actuation and subsequently the drug release were controlled by a manipulated communication system based on transmitter and receiver circuits, designed for wireless power transmission. Electromagnetic waves of 2 MHz were applied for power transmission. The wireless transmission works properly even on the range of 7 cm which is suitable distance for this chip getting implanted in the patient's body, near the cancerous organ.

Resume : Hepatocellular carcinoma is the sixth most common cancer and the second leading cause of cancer mortality worldwide. Thus, there is still a great need to develop new materials for efficient therapy of this type of cancer. One of the most promising approaches towards cancer therapy is merging different modalities resulting in higher efficiently in eradication of cancer cells. Much attention has been draw by bioinspired polymer polydopamine which has been recognized as extremely effective and biocompatible photothermal agent. This polymer has just recently entered the field of material chemistry, nanotechnology and biomedicine.[1,2] The versatility of polydopamine was demonstrated by the deposition of polydopamine on a variety of materials including hydrophobic Teflon, noble metals, silica, carbon nanotubes, magnetic and gold nanoparticles. In this talk, the merging of polydopamine with magnetic nanoparticles towards multifunctional nanocarriers for combined chemo- and photothermal therapy of liver cancer will be presented.[3] Glioma belongs to the most aggressive and lethal types of cancer. Malignant glioma is characterized by a poor prognosis and remains practically incurable despite aggressive treatment such as surgery, radiotherapy, and chemotherapy. Brain tumor cells overexpress a number of proteins that play a crucial role in tumorigenesis and may be exploited as therapeutic targets. One of such target can be an extracellular matrix glycoprotein - tenascin-C (TN-C). A down-regulation of TN-C by RNA interference (RNAi) is a very promising strategy in cancer therapy. This part of the talk will deal with combined gene and photothermal therapy of glioblastoma using mere polydopamine particles. ACKNOWLEDGMENTS: The research was financed by The National Science Centre (NCN), Poland under project number 2016/21/B/ST8/00477 and by the National Center for Research and Development under research programme LIDER/11/0055/L-7/15/NCBiR/2016. Radosław Mrówczyński is also grateful to the Foundation for Polish Science (FNP) for its support through a START scholarship. [1] R.Mrówczyński et. al. Polym.Chem. 2016, 65, 1288 [2] R. Mrówczyński Appl. Mater. Interfaces, 2018, 10, 7541 [3] R. Mrówczyński et al. Nanomaterials 2018, 8,170

Resume : Ferrites belong to a specific class of multifunctional materials. We have succeeded in synthesizing of cubic compounds in form of nanograins with varied composition Me(1+x)Fe(2-x)O4 (0.1< x< 1) using conventional precursors and organic environment under atmospheric-pressure conditions. Usage of different element allow to tune magnetic properties of nanostructures and therefore its usability including bioaplication. Number of examples will be given with description of properties and potential application.