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Smart biointerfaces for functional biomaterials

The symposium on “Smart Biointerfaces for Smart Biomaterials” aims to bring together scientists from universities, research institutes and industries to review the current frontiers in the strongly interconnected areas of Nanobiotechnology, Biointerfaces and Biomaterials. The symposium will focus on a set of interdisciplinary topics, spanning from advanced drug delivery systems to novel tissue engineering strategies, from smart diagnostic and nano-resolved imaging techniques to safety of nanomaterials.

Scope:

The field of Smart Biointerfaces is markedly interdisciplinary, bridging together bionanotechnologies, biomimetic devices, tissue engineering and biohybrid systems. The key to achieve important advances in the field of biointegrated devices resides in the successful integration of these technological and scientific areas. In this context, diverse but complementary contributions are needed on new biomaterials, multi-signal patterning methodologies, multiscale modelling, advanced characterization and processing technologies for the desired biomedical and biotechnological applications. Thus, the concept of Smart Biointerfaces is constantly gaining new valences. Chemical structures of the interfaces along with the electrical, mechanical and morphological properties at nanoscale appear equally relevant to drive the interactions between living and synthetic systems. A central aspect is then the ability to optimize the functional properties with high spatial resolution, creating materials that are able to control the interaction with the biological surrounding at the nanoscale thus guiding the responses of biomolecules, cells and tissues. Accordingly, by responding to changes in the biological environment, or transformation from one state to another in the presence of biological systems, functional biomaterials must not only improve device integration and control tissue regeneration, but also use controlled responses to power hybrid biodevices. In this view, the symposium will seek to integrate the experimental and theoretical research endeavours drawing the strengths from all the aspects of interface design and fabrication as well as characterization of the interactions at the material/biological systems interface.

Hot topics to be covered by the symposium:

  • Responsive biointerfaces
  • Small biointerfaces
  • Cell material interactions
  • Functional biomaterials
  • Drug delivery systems
  • Cell instructive materials
  • Tissue engineering scaffolds
  • Biomedical implants
  • Novel polymers and biopolymers
  • Stimuli and cell responsive materials
  • Biomolecules surfaces interaction
  • Bionterfaces engineering
  • Surface treatments for biomedical applications
  • Nanoparticles-biological interaction
  • Nano and micropatterning for biomedical application
  • Smart biohybrid materials
  • Porous and composite biomaterials
  • Biomedical Microsystems
  • Micro and nanosystems for biological recognition
  • Antibacterial surfaces
  • Blood- and tissue-material interactions
  • Modelling of cell material interaction and biological recognition

 

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Peptide and Protein Assemblies : Jian R Lu
13:30
Authors : Mariano Venanzi,1 Emanuela Gatto,1 Mario Caruso,1 Fernando Formaggio,2 Marta De Zotti2
Affiliations : 1 Dept. of Chemical Sciences and Technologies, University of Rome 'Tor Vergata', Italy. 2 Dept. of Chemistry, University of Padova, Italy.

Resume : Peptide-based materials have been finding application in very different fields, spanning from molecular electronics to antifouling coating to tissue engineering. Mimicking Nature, self-assembly is the most suitable strategy for obtaining peptide structures with nanometric control of morphology (bottom-up approach). The idea is to determine the morphology of supramolecular architectures by selecting peptide building blocks of well-defined secondary structure and adopt the proper procedures to propagate this information from the molecular to the nano and mesoscopic size (hierarchical self-assembly). To the aim, we synthesized oligo peptides rich in Cα-tetrasubstitued amino acids. Due to the restricted torsional angles available, peptides formed by such amino acids are constrained to attain a specific secondary structure, representing molecular building blocks of well-defined shape and size. Two examples of 2D peptide nanostructures will be discussed at the Conference: i) peptide self-assembled monolayers, covalently linked to gold substrates by a lipoic chain, and ii) peptide films obtained by Langmuir-Blodgett deposition. In both cases, the influence of the secondary structure on the aggregation propensity of the peptide building blocks and on the physical properties (density, electronic conduction, photophysics) of the peptide films will be thoroughly analyzed.

Q.1.1
14:00
Authors : G.M.L. Messina1, M. De Zotti2, A. Rapisarda1, F. Formaggio2 and G. Marletta1
Affiliations : 1 Laboratory for Molecular Surfaces and Nanotechnology, Dept. of Chemical Sciences, University of Catania, Catania, I-95125, Italy 2 Department of Chemistry, University of Padova, Padova, I-35131, Italy

Resume : Biomolecule-based materials that change properties in response to different local stimuli are increasingly being studied in the context of several applications, with particular attention to bioelectronics. Peptides are ideally suited for this purpose because of the range of distinct physical properties available from the amino acids. This diversity allows several non-covalent interactions including electrostatic (acidic and basic amino acids), hydrophobic, p-stacking (aromatic amino acids), hydrogen bonding (polar amino acids) as well as covalent (disulfide) bonds and steric contributions (strand directing amino acids). Crucially, these interactions depend by different factors such as ionic strength, pH and temperature. In this work, we study the properties of a particular peptide able to respond to specific external stimuli by adopting one of two well-defined conformations. The Trichogin GA IV here used, belongs to the family of peptaibiotics, antimicrobial peptides rich in the helix-inducer α-aminoisobutyric acid (Aib) residue. One of its analogs, in which four Lysines positive charged residues have been introduced, is able to reversibly switch its conformation between two, well-defined, different helical conformations in response to pH variations. In particular, it is shown that this peptide can contract and stretch in response to a short range of pH variation. In this contribution, we describe our latest results obtained from the study of stimuli-responsive smart surfaces, formed by gold substrates decorated with trichogin GA IV and its positively-charged analog. The peptides were anchored to the gold surfaces through a N-terminal lipoic acid moiety. The loading and the conformational switching properties of the surface-bound peptides were investigated by means of several techniques, such as Quartz Crystal Microbalance with Dissipation monitoring and Localized Surface Plasmon Resonance. Applications in the emerging field of the bio-inspired nanotechnology are suggested.

Q.1.2
14:15
Authors : Nicoletta Giamblanco, Nunzio Tuccitto, Gabriella Zappalà, Antonino Licciardello, Giovanni Marletta
Affiliations : Laboratory for Molecular Surfaces and Nanotechnology, Department of Chemical Science, University of Catania, viale A. Doria n° 6, 965125, Catania, Italy and CSGI

Resume : Characterization of protein adsorption from an aqueous solution onto a surface is an important element of research in surface science, from both fundamental and practical perspectives. During the last few years, we focused our attention on protein layer adsorption in terms of adsorbed amount, structure of the layer and adsorption kinetics. Spontaneous surface adsorption of proteins results in the formation of molecular films having peculiar thickness and structural conformation. We present an original study concerning the adsorption of human serum albumin (HSA) and lactoferrin (LF) onto Fe(II) and Cu(II) terpyridine-based complexes self-assembled on a gold surface. This contribution deals with an integrated characterization approach aimed to obtain detailed insights into the nature of the surface-induced conformational changes in co-adsorbed proteins. This approach involves time-of-flight secondary ion mass spectrometry measurements coupled with multivariate data treatment and parallel experiments using quartz crystal microbalance with dissipation monitoring (QCM-D). In particular, Principal Component Analysis (PCA) was used to identify similarities and differences in TOF-SIMS spectra of protein-based films and to classify the spectra into groups. PCA results shown that single-protein films can certainly be discriminated. Films prepared by mixed-component solutions revealed peculiar positions in the scores plot. The ToF-SIMS spectra of proteins adsorbed onto Fe(II) complex were clustered close to the HSA single-component layer. Similarly, films prepared on Cu(II) have surface mass spectra similar to that of the FN single-component film. On the other hand, QCM-D studies show that HSA exhibits a strong affinity with the Cu(II)-containing self assembled monolayer, whereas LF interacts much strongly with Fe(II)-based SAMs rather than with HSA. Consequently, we conclude that the PCA results on TOFSIMS data can be interpreted in terms of competitive multilayer adsorption. In other words, when allowing a mixed-component (HSA/FN) protein to interact with the metal complex monolayer, HSA competitively interacts with Cu(II)-based complexes and adsorbs on them. This stage is followed by the subsequent adsorption of FN on top of the HSA layer. By contrast, the low interaction of HSA with Fe(II) complex promotes the formation of a first layer of LF, followed by the adsorption of HSA on top of it. Sequential adsorption QCM-D experiments confirmed such interpretations of the peculiar physico-chemical process.

Q.1.3
14:30
Authors : Kristin Hyltegren, Tommy Nylander, Mikael Lund, Marie Skepö
Affiliations : Division of Theoretical Chemistry, Lund University; Division of Physical Chemistry, Lund University; Division of Theoretical Chemistry, Lund University; Division of Theoretical Chemistry, Lund University

Resume : Histatin 5 is a saliva protein that acts as the first line of defence against oral candidiasis caused by Candida Albicans. The antimicrobial activity has been ascribed to the high content of basic amino acids. Histatin 5 also participates in the formation of a protective layer on smooth tooth surfaces, and thereby prevents microbial colonization and stabilizes mineral–solute interactions. Thus, the adsorption of histatin 5 to surfaces in the oral cavity is important for our oral health. Our aim is to understand how the lack of structure of histatin 5 in solution relates to its function in the adsorbed state. The focus of this study is to see how pH, ionic strength and charge regulation of the protein affects surface adsorption. For this purpose a combination of ellipsometry and coarse-grained Monte Carlo simulations have been used. By applying the Langmuir adsorption isotherm, a surface coverage could be calculated from the single-protein simulation results, enabling a direct comparison with experiments. The results show that inclusion of an attractive interaction of 2.9 kT per amino acid is necessary to get close to experimental results. The main reason is that our coarse-grained model overestimates the entropy of the free histatin 5 chain. The rest of the potential represents van der Waals interactions, hydrogen bonding and possibly charge regulation of the surface. Preliminary results will be shown for the influence of charge regulation of the surface.

Q.1.4
14:45
Authors : Jiqian Wang, Songsong Chang, Naidong Zhang, Hai Xu, Jian R Lu
Affiliations : Jiqian Wang, Songsong Chang, Naidong Zhang, Hai Xu; China University of Petroleum (East China); Jian R Lu; University of Manchester

Resume : Short peptide amphiphiles have been explored as effective nanobiomaterials in applications ranging from controlled gene or drug release, tissue repair, biomineralization to 3D cell culture. Some short peptide amphiphiles have shown antimicrobial and anticancer activity, and have already been used as new antibiotics, and also are tried as anticancer drugs. The assembly of short peptide amphiphiles is driven by a series of weak forces, including hydrogen bonding, electrostatic interaction, hydrophobic interaction, Van der Waals interaction, aromatic interaction. The polymorphisms of self-assembly could be tuned by regulating these non-covalent forces. Short peptide amphiphiles usually self-assemble into one dimentional nanostructures including nanotubes, nanofibrils, nanoribbons and vesicles. The self-assembly process and morphology are dictated by molecular structures and solution environment as well. As reported by many reseachers, the roles of leucine and isoleucine residue are significantly different in short peptide amphiphile self-assemblies or protein structures although they are isomers with very similar molecular strutures. We have designed a series of short peptide amphiphiles with lysine as the hydrophilic head, leucine and isoleucine as the hydrophobic chain. These amphiphiles are cataloged into three groups according to the number and position of leucine and isoleucine residues to study the effects of leucine and isoleucine in self-assembly. Effect of solvent on the morphology was also studied by changing the solvent polarity. Our results showed that by reducing solvent polarity the assemblies of I4K and I3LK changed from nanofibrils to nanoribbons, but those of ILI2K、I2LIK and LI3K assemblies didn’t change. The reducing the solvent polarity enhanced the hydrogen bonding interactions between β strands, and weakened the hydrophobic interaction, and thus the β-sheet structures of short peptide amphiphiles became more rigid and was not easy to distort. As a result, they formed less twisted nanoribbons instead of highly twisted nanofibrils. When leucine was located in the N terminal of the molecules, the short peptide amphiphiles usually assembled into nanofibrils in 10% acetonitrile solution. While the leucine residues were in the middle of short peptide amphiphiles, their self-assembly morphologies were often nanoribbons. Combining with TEM, AFM, CD and SANS results, we deduced that the morphology transformation of short peptide amphiphile assemblies was determined by the twisted ability of peptide molecules in the β-sheet structure. If the β-sheet structure was easy to be distorted, the self-assembles usually formed nonofibril structure, instead of nanoribbon structure. Leucine residues at N-terminal promoted the distortion of β-sheet structure and thus made the self-assemblies into naofibrils structure. On the contrary, the leucine residues in the middle of molecules retarded the distortion, and as a result, they formed nanoribbon structures.

Q.1.5
15:00 Break    
 
Functionalised and responsive surfaces I : Giovanni Marletta
15:30
Authors : John Webster
Affiliations : ISIS neutron facility, Rutherford Appleton Laboratory, STFC, Chilton, Didcot, Oxon OX11 0QX

Resume : In this invited talk, I will first introduce the unique capability of neutron reflection aided by the isotopic contrast variation via H/D exchanges at studying protein adsorption, followed by illustrating the main structural features that can be revealed at different interfaces. I will then show how a typical globular protein interacts with different model surfactants leading to structural unfolding and changes in interfacial structure and composition, emphasising the importance of the appropriate isotopic constrasts to be used. Using IgG as examples, I will how neutron reflection can be used to study its adsorption and desorption at the air/water and solid/water interfaces, thereby improving our understanding about how surface and interfacial adsorption could cause structural destabilisation.

Q.2.1
16:00
Authors : MUTSCHLER Angela, SCHAAF Pierre, LAVALLE Philippe and VRANA Nihal Engin
Affiliations : Institut National de la Santé et de la Recherche Médicale, INSERM Unité 1121, 11 rue Humann, 67085 Strasbourg, France

Resume : -

Q.2.2
16:15
Authors : Li Deng, Yurong Zhao, Peng Zhou, Hai Xu*1, and Yanting Wang*2
Affiliations : 1) Centre for Bioengineering and Biotechnology, China University of Petroleum (East China) 2) Institute of Theoretical Physics, Chinese Academy of Sciences

Resume : Nanostructures self-assembled by cross-β peptides are with highly ordered hierarchical structures and high mechanic properties have potential applications in biomaterials.and formed by lateral stacking of cross-β sheets. Quantitatively studying the intermolecular force and driving forces in of different hierarchical structures at the microscopic atomistic level is essential for understanding the mechanism to form various morphologies and nanomechanics of nanostructures self-assembled by various kinds of peptides. In this work, we investigate the thermodynamics of intra-sheet and inter-sheet structures self-assembled by KIIIIK with the umbrella sampling technique applied toby atomistic molecular dynamics simulations. Combining combining steered molecular dynamics simulation with umbrella sampling., the potential of mean forces (PMF) of both structures have been calculated. It is found that The the mechanical properties of intermolecular bond stiffness intra-sheet and inter-sheet structure are highly anisotropic and their intermolecular bond stiffness of are 5.58 N/m and 0.32 N/m respectively. By analyzing the entropic and enthalpic contributions for association of intra-sheet and inter-sheet structure, we find that the mechanical anisotropy comes from that the difference between their driving forces to keep the equilibrated structures stable. The enthalpy keeps intra-sheet structure stable, but the entropy keeps inter-sheet structure stable. Furthermore, the driving forces in the association process of intra-sheet and inter-sheet structure are different due to that the van der Waals and electrostatic interactions between peptide and peptide and that between peptide and solvents in different shells have play different roles in association of these structuresthe processes. Especially, the interactions between peptide and solvent in hydration shell have significant effects on the peptide self-assembly. The For the intra-sheet structure of KIIIIK is stabilized by hydrogen bond interaction, so the electrostatic interaction between peptide and peptide dominate the association of strands but the electrostatic interaction between peptide and the solvent in the hydration shell is disadvantageous to association of strands. However, for the inter-sheet structure of KIIIIK is stabilized by hydrophobic interaction, so the van der Waals (VDW) interaction between peptides promotes the association of sheets and the interaction between peptide and the solvent in the hydration shell also is advantageous to promote the aggregationassociation of sheets.

Q.2.3
16:30
Authors : Alexandra Tibaldi, Vincent Noel, Marion Woytasik, Benoit Piro, Laure Fillaud, Giorgio Mattana
Affiliations : University Paris Diderot, ITODYS, UMR 7086 University Paris Saclay, IEF, UMR 8622

Resume : Highly sensitive, robust and miniaturized transduction systems could overcome the typical problems associated with the early diagnostic of diseases that requires the simultaneous identification and quantification of several biomarkers (proteins) at the trace level. A new kind of organic field-effect transistor architecture, the so-called "Electrolyte-Gated Field Effect Transistor" (EGOFET) offers a new analytical tool with an extremely high sensitivity of its electrical characteristics toward any physicochemical phenomenon occurring in the vicinity of the gate and / or transistor channel. To perform simultaneously a precise quantification of proteins at the trace level (up to 10-12 M), we adapt the standard ELISA (Enzyme-Linked Immunosorbent Assay) setup by replacing the conventional enzyme label (i.e., usually an enzyme producing a chromophore detected by UV-Vis spectroscopy) by an acetylcholinesterase. This enzyme converts the acetylthiocholine in thiocholine (TC). The TC has a thiol group that spontaneously chemisorbs on the EGOFET gate (Au), leading to a drastic change of the transistor characteristics. The proof of concept of an ELISA read-out performed using an organic field effect transistor will be presented as well as the current device figures of merit. This work paves the way to the development of new ultra-sensitive protein sensors that are compatible with the integration of microfluidic systems as well as large-scale and low-cost fabrication processes.

Q.2.4
16:45
Authors : Tugba Isik, Nesrin Horzum, Ü. Hakan Yildiz, Mustafa M. Demir
Affiliations : Tugba Isik - Materials Science and Engineering Department, Izmir Institute of Technology, Turkey ; Nesrin Horzum - Biomedical Engineering, Izmir Katip Çelebi University, Turkey ; Ü. Hakan Yildiz - Chemistry Department, Izmir Institute of Technology, Turkey ; Mustafa M. Demir - Materials Science and Engineering Department, Izmir Institute of Technology, Turkey

Resume : Biomarkers have been used in clinical diagnosis. They are measurable indicators for the physiological state of the body. Recent studies demonstrate that the change in the level of biomarkers indicates the early stages of cancer or cardiovascular diseases. Therefore, the separation and concentration of biomarkers for further analysis have high potential in early detection. This study asserts a convenient approach to develop affinity membranes for the separation of nucleic acid biomarkers. The separation efficiency of membranes was examined with bovine serum albumin (BSA) as model protein and single stranded DNA (ss-DNA) as biomarker. The membranes made of poly styrene (PS) and poly (methyl methacrylate) (PMMA) were fabricated by electrospinning. The sorption efficiency of membranes suggests that separation of ss-DNA and proteins are possible. The preliminary results revealed that electrospun PS membranes (after surface modification) are promising for BSA uptake with 111 mg g-1 sorption capacity. Also, PS membranes show better affinity to BSA molecules by hydrophobic interactions. In the model mixture of BSA and ss-DNA, by virtue of anti-fouling property of BSA, ss-DNA cannot be held on the surface and the ratio of BSA/DNA in eluted solution introduced three-fold decrease by increasing the adsorbed BSA on the membranes. The proposed technology promises fast and effective separation of biomarkers from both blood and body fluids.

Q.2.5
17:00
Authors : A. Varesano1, C. Vineis1, D.O. Sanchez Ramirez1, R.A. Carletto1, F. Truffa Giachet1, S. Spriano2, S. Ferraris2, L. Rimondini3, N. Bloise4, L. Visai4
Affiliations : 1CNR-ISMAC, Istituto per lo Studio delle Macromolecole, BIELLA, Italy; 2Politecnico di Torino, DISAT, TORINO, Italy; 3Università del Piemonte Orientale, NOVARA, Italy; 4Università di Pavia and S. Maugeri Foundation, IRCCS, PAVIA, Italy

Resume : Electrospinning is a simple method to produce nanofibres with high specific surface. Keratin is a biocompatible and biodegradable protein [1], it supports fibroblasts [2] and osteoblasts [3] growth. Moreover, electrospun keratin-based nanofibers (EKNs) were made insoluble to water by a thermal treatment inducing cross-linking [4]. For dental applications, pure keratin has been electrospun from solutions of formic acid to deposit nanofibers onto titanium substrates. The aim is the improvement of soft tissue adhesion avoiding epithelial down-growth and bacterial infiltration on the collar. Water stability of EKNs was evaluated by 28 days soaking. EKNs were found to greatly improve fibroblast cells growth already at 24h. In bone regeneration, nano-hydroxyapatite (HAp) was used in composite EKNs. Unfortunately, HAp particles are not stable in acids; therefore, water was used as solvent to produce HAp-EKNs. Better processability was attained by blending keratin with polyethylene oxide (PEO). PEO is an amphiphilic and water-soluble polymer added as a sacrificial material. HAp-EKNs were subjected to heating and washing to obtain pure keratin nanofibres. HAp-EKNs have been tested for osteoblasts cells adhesion and growth, showing biocompatibility by cell viability assay and SEM. [1] Yamauchi et al. J Biomater Sci Polym Ed 9 (1998) 259. [2] Tachibana et al. J Biotechnol 93 (2002) 165. [3] Tachibana et al. Biomaterials 26 (2005) 297. [4] Varesano et al. J Appl Polym Sci 131 (2014) 40532.

Q.2.6
17:15
Authors : Muling Zeng, Anna May-Masnou, Anna Roig, Anna Laromaine
Affiliations : Institut de Ciència de Materials de Barcelona, Campus UAB, 08193 Bellaterra, Spain.

Resume : Cellulose from microbial origin, commonly known as bacterial cellulose (BC), is becoming a commodity material since it incorporates desirable structural properties for biomedical applications. Here, we present the production and characterization of bacterial cellulose (BC) films of less than a hundred microns thick produced by Gluconacetobacter bacteria. These thin films are processed using three different drying methods: 1) room temperature, 2) freeze drying and 3) supercritical drying. The different processes confer the cellulose films a hierarchical porous network, high purity and crystallinity, flexibility and strength (high Young modulus at room and elevated temperatures) and large water holding capacity that can be tailored and controlled for different applications. In this work, we used BC films as a platform to incorporate magnetic functionality by the incorporation of iron oxide nanoparticles and gold nanoparticles as potential catalysts. Using the microwave-assisted method in a rapid and cost effectively manner, we magnetically coated the whole structure of our BC films uniformly. By evaluating different precursor´s concentrations and taking advantage of the different drying methods, we achieved magnetic bacterial films with different magnetic strength. We obtained magnetic, flexible and robust BC composites within a few minutes in a clean, easy and reproducible method. We present different options to increase the complexity of these BC films, including the attachment of metallic or oxide nanoparticles, or patterning the BC films with different hydrophobic domains.

Q.2.7
17:30
Authors : Aharon Gedanken
Affiliations : Department of Chemistry, Bar-Ilan University, Ramat-Gan, 52900, Israel.

Resume : Sonochemistry is an excellent technique to coat functional nanomaterials on various substrates, and imparting new properties to the substrates. Last year I have presented the results of an EC project aimed at making the Hospital a safer place by the sonochemical coating of all the textiles with ZnO, CuO NPs, and especially Cu0.89Zn0.11O which kills bacteria about 10,000 times than the former oxides. These NPs killed not only sensitive gram-negative E. Coli (strain 1313) as well as the gram-positive Staphylococus aureus (strain 195) bacteria very efficiently, but also resistant bacteria such as MRSA. We have extended the previous studies and included some edible organic nanoparticles such as vanillin nanoparticles (~ 50nm in size), raspberry ketone (RK) nanoparticles (~40nm in size) and camphor nanoparticles (width ~30nm, length ~40nm in size) on fabrics and on polypropylene surfaces. Their biocidal activity against Escherichia coli (E. coli), Salmonella paratyphi A (S. paratyphi A) and the yeast Candida albicans (C. albicans) cultures was demonstrated. In addition we have sonochemically coated Cu0.89Zn0.11O on Silicon Catheters, and artificial teeth. Very good results were obtained in attempting to inhibit the growth of Biofilm on these surfaces. In vivo experiments in mice in which coated catheters were inserted showed good results in avoiding biofilm formation. Figure 1 presents the rabbits and the catheters installed in their bodies.

Q.2.8
17:45
Authors : T. Aschl, A. Moraillon, F. Ozanam, P. Allongue, A.-C. Gouget-Laemmel
Affiliations : Physique de la Matière Condensée, Ecole Polytechnique-CNRS, France

Resume : Aptamer-based biochips are well suited to the identification of targets like proteins, toxins or bacteria because they allow rapid and sensitive detection [1]. The prerequisite for achieving selective target recognition is depositing a bioreceptive organic layer, whose chemical properties allow the controlled immobilization of suitable aptamers (probes) and also minimize non-specific target adsorption. In this work we aim at detecting Ochratoxin A (OTA), a well-known mycotoxin which has nephrotoxic, teratogenic and immunotoxic effects, using fluorescence measurements. The biochip is composed of an Al-on-glass back reflector covered by a thin film of amorphous Si-C alloy [2], which is adjusted to obtain constructive interferences at the film surface. A carboxyl-terminated organic layer is then grafted to the surface via stable Si-C bonds. Surface activation with succinimidyl ester groups is performed for the amidation of the well-known 36mer AntiOTA aptamer. ATR-IR characterizations of the assembly, until aptamer / OTA duplex formation, will be presented at the conference to discuss the organic layer composition and estimate the surface concentration of immobilized OTA in various physiological media. Complementary fluorescence assays will also be presented. Preliminary results indicate successful interaction of OTA with the immobilized aptamers. [1] F. Radom et al. Biotechnol. Adv. 2013, 31, 1260. [2] L. Touahir et al. Biosens. Bioelectron. 2009, 25, 952.

Q.2.9
 
Poster Session : -
18:00
Authors : BEY Houda*, GTARI Wala, ASCHI Adel and OTHMAN Tahar
Affiliations : Laboratoire de Physique de la matière Molle et de la Modélisation Electromagnétique, Faculté des Sciences de Tunis, Université El Manar, 2092 Campus Universitaire, Tunis, Tunisia

Resume : Small angle light scattering (SALS) is a new technique for structural measures in a large size range, which is suitable for the study of soft matter [1]. It is a method of light scattering that bridges the gap between the broadcasting techniques standard light as dynamic or static Light Scattering (DLS and SLS) and microscopy. The sample is illuminated with an expanded beam of coherent light and a camera (CCD) monitors the intensity of the light transmitted. The image shows speckle fluctuations, which are captured by the speckle[2]. In this work, we will study the influence of physicochemical parameters (temperature, pH, pressure ...).We developed complex coacervation (protein and polyelectrolyte) that leads to the formation of microcapsules[3]. The understanding the mechanisms of formation of coacervates in different environmental conditions is essential if we are to predict and improve techno-functional properties of coacervates. Indeed, it has been shown that the functional properties of the complexed biopolymer were higher than those of the biopolymers alone and that the areas of use of these complex macromolecular assemblies were multiple. This work is widely used in many industrial applications as the development of new multifunctional food ingredients include the purification of biological molecules by precipitation, micro- or nano-encapsulation of active substances in pharmacy, medicine or cosmetology, synthesis of therapeutic vectors synthesis biomaterials biocompatible or development of biosensor. Keywords: Small Angle Light Scattering, Complex, Coacervats, Microencapsulation Référence: [1]Cipelletti L., Carpineti M.,Giglio M, Two-color cross-correlation in small-angle static light scatterin,Physical Review,53,1997. [2]Bhatia S.R, Ultra-small-angle scattering studies of complex fluids.Current Opinion in Colloid & Interface Science: 9, 404 – 411,2005. [3] Jain A.,Thakur D.,Ghoshal G. ,Katare O. P. , Shivhare U. S.,Microencapsulation by Complex Coacervation Using Whey Protein Isolates and Gum Acacia: An Approach to Preserve the Functionality and Controlled Release of β-Carotene,Food Bioprocess Technol ,8:1635–1644,2015.

Q.P.1
18:00
Authors : GTARI Wala*, BEY Houda, ASCHI Adel and OTHMAN Tahar
Affiliations : Laboratoire de Physique de la matière Molle et de la Modélisation Electromagnétique, Faculté des Sciences de Tunis, Université El Manar, 2092 Campus Universitaire, Tunis, Tunisia

Resume : The intracellular environment represents an extremely crowded environment, with a complete lack of unoccupied space and a limited amount of free water. Proteins are surrounded by several other molecules of different chemical nature, and this crowded environment can considerably modify their behavior. To mimic the real cells, we used a "crowding agent" to study the macromolecular crowding. The literature revealed that macromolecular crowding might affect protein structure, folding, shape, conformational stability, binding of small molecules, enzymatic activity, protein-protein interactions, and pathological aggregation. The modification of the conformation of proteins in the absence and the presence of high molecular crowding agent was characterized by Small Angle and Dynamic Light Scattering (SLS and DLS). The results show that the molecular crowder (polysucrose 400) has a strong effect on the chemical denaturation process of both proteins. The variation of the measured hydrodynamic radius is attributed to the enhanced effect of the crowder agent in agreement with the excluded volume effects. The large size of the molecular crowder is shown to play a key role for the protein conformation in a native or unfolded state. This work has addressed the general effect on the unfolding process of protein in the presence of Polysucrose 400. The excluded volume effect is due to the presence of inert macromolecular crowding on unfolded proteins. The enhanced denaturation process on proteins was very important since it was accelerated at certain concentrations of poly400. We supported the hypothesis which predicts that the crowder size and shape have an important factor that governs the effect of molecular crowding on proteins.

Q.P.2
18:00
Authors : Benedetta Castroflorio, Alessandro Rapisarda, Grazia ML Messina, Carmela Bonaccorso, Valentina Spampinato, Domenico Sciotto, Giovanni Marletta
Affiliations : Department of Chemical Sciences, University of Catania, Catania, I-95125, Italy

Resume : Biofouling is an important issue affecting numerous applications ranging from biomedical implants to biosensors. Therefore, there is a constant need to develop versatile, convenient and cost-effective autifouling strategies in healthcare. Among the others, surface functionalization with antifouling structure is a convenient way for giving adhesion resistance. Since to construct protein resistant surfaces would require to have no net charge and/or polar functional groups, a strategy involving the surface immobilization of zwitterionic units has been investigated, as a potentially attractive method to prepare antiprotein fouling surface. Potential optimal candidates to build up effective antifouling surfaces for in vivo application are natural amino acids, in their zwitterionic form. In this study we describe the anchoring of calix[4]arene-crown-5 self-assembled monolayer on gold surface, to be used as containers of aminoacids, and the interaction between surface-anchored calix and amino acid residues in aqueous environment. The amino acid inclusion in the calix was studied by using Quartz Crystal Microbalance with Dissipation monitoring (QCM-D) technique. As the amino acids exhibit pH-dependent behavior, the pH values in the aqueous environment strongly influence the molecular recognition process in solution due to the combined effect of electrostatic and hydrophobic interactions. In particular, it was found that the inclusion efficiency, while depending in a minor way on the hydrophobicity, appear to critically depend on the flexibility of the amino acids, i.e., from the matching of the steric constrains and conformational change of the including amino acids. The antifouling properties of the functionalized zwitterionic surfaces have been tested against Human Serum Albumin and Lysozyme.

Q.P.3
18:00
Authors : C. Vineis1, A. Varesano1, C. Tonetti1, D. O. Sánchez Ramírez1, R. A. Carletto1, S. Ortelli2, M. Blosi2, A. L. Costa2
Affiliations : 1CNR-ISMAC, Institute for Macromolecular Studies – National Research Council of Italy, C.so Giuseppe Pella 16, I-13900 Biella, Italy; 2CNR-ISTEC, Institute of Science and Technology for Ceramics – National Research Council of Italy, Via Granarolo 64, I-48018 Faenza (RA), Italy

Resume : Multi-component organic/inorganic nanofibers were produced by electrospinning water solutions of keratin extracted from wool containing nanosols of titanium dioxide or metal silver. The resulting hybrid keratin-based nanofibers were made insoluble to water by treatments at high temperature. Nanofibers were observed by SEM before and after the heat treatments, and after the contact with water. Energy Dispersive X-ray analysis was performed with the aim of confirm the presence of inorganic nanoparticles. Finally, the functional properties (antibacterial property and photo-catalytic activity) of the nanoparticles embedded into the electrospun nanofibers were quantified on treated samples. The TiO2 reactivity was tested in both cases under UV light irradiation. Antibacterial tests were carried out using E. coli following the ATCC 100 Test Method on nanofibers treated with silver or TiO2. Both electrospun hybrid nanofibers showed excellent antibacterial properties confirming that the nanoparticles can exert their functions even if they are embedded in keratin nanofibers. Photocatalytic tests based on model reaction of Rhodamine B dye degradation showed promising photocatalytic property for the hybrid TiO2/keratin electrospun nanofibers. The results demonstrated that electrospun keratin nanofibers doped with active nanophases preserve nanoparticle properties, allowing the design of highly versatile hybrid multifunctional media.

Q.P.4
18:00
Authors : A. Vladescu1, M. Braic1, A.Gherghilescu2, A.Kiss1, C. M. Cotrut2, I.Titorencu3, V.Jinga3, V.Braic1
Affiliations : 1National Institute for Optoelectronics – INOE 2000, 409 Atomistilor Str., RO77125, Magurele, Romania 2University Politehnica of Bucharest, 313 Splaiul Independentei, RO 060042, Bucharest, Romania 3Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy, 8 B.P.Hasdeu, Bucharest, Romania

Resume : Hydroxyapatite coatings enriched with Ti were prepared as a possible candidate for biomedical applications especially for implantable devices that are in direct contact to the bone. The coatings were prepared by RF magnetron sputtering method on Ti based alloy using pure hydroxyapatite and TiO2 targets. The content of Ti was modified by changing the RF power fed on TiO2 target. The formation of the hydroxyapatite compound was not influenced by the addition of Ti. The Ca/P ratio of the Ti doped hydroxyapatite coatings was ranged from 1.64 to 1.68, being close to the stoichiometric hydroxyapatite coating. The roughness of the doped hydroxyapatite coatings were increased by increasing the RF power on TiO2 cathode. The in vitro corrosion performances of uncoated substrate were significantly improved by hydroxyapatite coating with or without Ti addition. The Ti additions led to an increase in cell viability of hydroxyapatite coatings, after 5 days of culture. The scanning electron microscopy showed that more cells were seen on the surface of hydroxyapatite enriched Ti than those observed on the surface of the uncoated Ti6Al4V alloys or hydroxyapatite without Ti addition.

Q.P.5
18:00
Authors : D. Arvaniti 1,2, V. Karagkiozaki 1,2, A. Papamichail 1, L. Tzounis 1, Th. Choli-Papadopoulou 3, S. Logothetidis 1
Affiliations : 1 Lab for Thin Films - Nanobiomaterials, Nanosystems & Nanometrology (LTFN), Νanomedicine Group, Physics Department, Aristotle University of Thessaloniki (A.U.Th.), Greece 2 BL NanoBiomed P.C, Thessaloniki, Greece 3 Biochemistry Lab, Department of Chemistry, A.U.Th., Greece

Resume : Gold nanoparticles (AuNPs) find numerous applications in the field of Nanomedicine, due to their unique physical properties, ease of functionalization and good biocompatibility. Biomolecules such as proteins could be easily attached onto AuNPs via electrostatic or other physical adsorption. The scope of this work is synthesis, characterization and functionalization of Turkevich AuNPs with chitosan and streptavidin respectively for attachment of antibodies and biotinylated antibodies onto the surface of the functionalized AuNPs for further utilization in biosensing applications. AuNPs were synthesized using the Turkevich method with some modifications and addition of chitosan and streptavidin followed. Their sizes and zeta potential were measured via Dynamic Light Scattering whereas their morphology and surface topography were investigated via Atomic Force Microscopy (AFM). UV/visible Spectroscopy and X-ray Photoelectron Spectroscopy (XPS) were used for analyzing their surface chemistry. The results indicate formation of Turkevich AuNPs with negative z-potential and uniformity, whereas high absolute value of z-potential indicates stability of the NPs. XPS and UV/visible spectrums are consistent with the value of pure Au and in accordance with bibliography. AFM and z-potential measurements suggest successful surface modification with chitosan and attachment of streptavidin onto the surface of AuNPs. Size, Peak-to-Peak and Root Mean Square values increase and z-potential changes dramatically after the surface modification. Hence, the synthesized AuNPs constitute a promising agent for further attachment of antibodies. Acknowledgements: This work has been partially supported by the NanoCardio Project “Nanomedicine for Advanced, Bioactive/ mimetic materials for Cardiovascular Implants”, funded by GSRT Greece& EC

Q.P.6
18:00
Authors : Nirmalya Tripathy,1 Rafiq Ahmad, 2 Yoon Bong Hahn,2 Gilson Khang1
Affiliations : 1Department of BIN Fusion Technology, Department of Polymer-Nano Science & Technology and Polymer BIN Research Center, Chonbuk National University,567 Baekjedaero, Deokjin-gu, Jeonju561-756, South Korea; 2School of Semiconductor and Chemical Engineering, and Nanomaterials Processing Research Center, Chonbuk National University, 567 Baekjedaero, Deokjin-gu, Jeonju561-756,South Korea

Resume : Intelligently designed surface nano-architecture provides defined control over the behavior of biomolecules and cells at the solid-liquid interface. Well-designed, mono-dispersed sodium-doped zinc oxide nanoparticles (Na-ZnO NPs) were prepared through wet chemical route at low temperature, and further formulated as paint to achieve thin film on glass surface. Surface morphological characterizations of the as-coated glass surface shows a uniform film thickness (~100±10nm) with homogeneous distribution and good attachment of nanoparticles to the glass surfaces. Photoluminescence spectra of Na-ZnO NPs exhibit exciton recombination emission along with deep and shallow trap emissions, attributed to oxygen deficiency associated with Na-dopant content. Antibacterial and antibiofilm assay showed remarkable reductions in bacterial growth and biofilm formation (Escherichia coli and Staphylococcus aureus) especially upon UV activation by reducing recombination of electrons/holes, compared to that of uncoated glass. The investigated mechanism reveals that the nanoparticles efficiently penetrate and generate intracellular reactive oxygen species (ROS) inside bacteria, which eventually enhances lipid peroxidation and cause cell death. Highlighting the superior efficacy of Na-ZnO NPs over ZnO NPs, this state-of-the-art design of Na-ZnO NPs based paint promises wide applicability in biomedical and industrial arena.

Q.P.7
18:00
Authors : R.A. Picca, M.C. Sportelli, R. Quarto, N. Ditaranto, A. Valentini, N. Cioffi
Affiliations : R.A. Picca: Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70126 Bari, Italy; M.C. Sportelli: Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70126 Bari, Italy;R. Quarto: Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70126 Bari, Italy; N. Ditaranto: Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70126 Bari, Italy; A. Valentini: Dipartimento Interateneo di Fisica “M. Merlin”, Università degli Studi di Bari “Aldo Moro”, Bari, Italy; N. Cioffi: Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70126 Bari, Italy

Resume : Copper-based nanoantimicrobials are often proposed as suitable tools to control/inhibit the growth of undesirable pathogens [1]. In our laboratory, we developed a successful approach for the electrochemical synthesis of highly stable copper nanoparticles (NPs) with proven bioactivity [2], in the presence of tetraalkylammonium salts as stabilizers. In this way, CuNPs can be employed for the modification of industrial products in order to confer them peculiar antimicrobial properties. Diluted solutions of CuNPs were used as impregnation baths for samples of polyurethane foams, used for matrasses. Two different stabilizers were chosen for CuNPs production: tetrabutylammonium chloride and tetraoctylammonium chloride, differing in the alkyl chain length. All the materials treated with CuNPs were characterized morphologically and spectroscopically. Pristine CuNP colloids were characterized by transmission electron microscopy. X-ray photoelectron spectroscopy was used to evaluate the surface chemical composition of pristine and treated polyurethanes. Copper ion release was investigated by Atomic Absorption Spectroscopy to correlate process parameters and release properties. The total amount of released Cu2 was found to be a function of alkyl chain length of the chosen stabilizer, as well as of CuNP loading into the final composite material. [1] A.P. Ingle et al., Appl. Microbiol. Biotechnol. 98 (2014) 1001. [2] “Nanomaterials for metal controlled release and process for their production” N. Cioffi, N. Ditaranto, L. Sabbatini, L. Torsi, P.G. Zambonin, EP n. 2123797B1.

Q.P.8
18:00
Authors : D.M. Vranceanu1, A. Vladescu2, M. Dinu2, A.I. Gherghilescu1, M. Tarcolea1, C.M. Cotrut1
Affiliations : 1University Politehnica of Bucharest, 313 Independentei Street, Bucharest, Romania 2National Institute for Optoelectronics, 409 Atomistilor Str., Magurele, Romania

Resume : In order to improve the bioactive behaviour of titanium, nowadays one major direction is to modify the surface by coating its with bioactive films. Formation of hydroxyapatite (HAp) coatings on Ti implants combines the favourable properties of both materials. Electrochemical deposition (ELD) is a cost-effective and versatile technique, in which the process parameters can be well controlled thought adequate conditions of the process. In this study, HAp was prepared by electrodeposition using a standard three electrode cell set-up in which Ag/AgCl was used as reference electrode, platinum electrode as counter electrode and the sample (Ti substrate) as working electrode. The electrolyte was prepared by dissolving Ca(NO3)2 and NH4H2PO4, in ultrapure water. A Potentiostat/Galvanostat operating in potentiodynamic mode was employed to a cathode potential of -1.4 V vs Eref for 2 h, at different temperatures in order to obtain the HAp coatings. The obtained HAp coatings have been characterized as follows: phase identification, morphology, chemical composition, roughness, contact angle and corrosion resistance. The linear polarization technique was used to obtain the polarization resistance (Rp), corrosion current densities (icorr) and corrosion rates (CR). As a conclusion it can be said that the temperature has influenced not only the morphology but also the corrosion resistance. A more stable behaviour being registered for the samples obtained at a higher temperature.

Q.P.9
18:00
Authors : Giuseppe Lanza
Affiliations : Dipartimento di Scienze del Farmaco, Università di Catania, Viale A. Doria 6, 95125 Catania (Italy)

Resume : The interaction of water with biomolecules is at the forefront of biophysical research since water modulates molecular structures and hence all fundamental functions related to life. Simple peptides dissolved in an aqueous environment have attracted great attention as proof of concept molecules to understand molecular properties and water-biomolecule interactions. In this context, the study has been devoted to DFT quantum chemical computations on the cationic Ala3H(nH2O) and neutral Ala3(nH2O) (n up to 40) complexes in a bottom-up fashion. The approach has been developed to model hydration effects on the molecular properties of trialanine in various form. Following simple and intuitive rules to arrange water molecules around the peptide, geometry optimization allowed us to find four minima for each form of trialanine corresponding to the unfolded extended (beta) and polyproline II (PPII) conformations. The peptide is incorporated into the network of hydrogen bonds of interfacial water molecules and it has been shown a more efficient intermolecular hydrogen bonding in the PPII arrangement with the following relative electronic energy stability beta-beta < beta-PPII = PPII-beta < PPII-PPII. The conformational entropy term proceeds in the reverse direction, thus these changes compensate in a way that leads to small changes in Gibbs free energy. These findings agree with experimental data which report an equilibrium between these conformers modulated by temperature.

Q.P.10
18:00
Authors : Yong-Tae Kim, Ji Hun Kim, Yong Ho Kim
Affiliations : Yong-Tae Kim, Ji Hun Kim, Yong Ho Kim, SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon, 440-746, South Korea; Yong Ho Kim, Department of Chemistry, Sungkyunkwan University, Suwon, South Korea

Resume : Supramolecular protein assembly is an ubiquitous process that physical and chemical principles of Mother Nature have achieved great complexity and stability to build up architectures of a life in molecular level. There have been so many trials of understanding how to construct supramolecular protein assembly and also a lot of successes have been built up. The stacked principles have recently been applied for designing the hybrid nanoparticle-protein superstructures that can show unique physical properties and potential application in the areas of plasmonics, molecular sensing, and nanoscale electronics. Here we show an unique nanostructure of self-assembled peptides that would allow for the wrapping of single walled carbon nanotubes (SWNTs) with specific helicities. We also demonstrate that the selection rule whereby precisely controlling position of functionality within such superstructures constrains a variety of arrangement of bimetallic AuPt nanoclusters for the purpose of enhancing catalytic activity of oxygen reduction reaction (ORR). The elected positions of functionality from the selection rule precisely alter their arrangement and density of catalytic nanoparticles and result in a significant change on ORR activity and durability. The remarkable electrochemical property of nanoparticle-protein-SWNT superstructures corresponding to controlling of interparticle distance, particle size and alloy composition of single-phase nanoparticles suggests a route to the construction of new functional protein nanomaterials tailored to unique energy applications.

Q.P.11
18:00
Authors : Jiyoung Nam, Yong-Tae Kim, Yoo Young Ahn, Nam Hyeong Kim, Ji Hun Kim, One-Sun Lee, Yong Ho Kim
Affiliations : Jiyoung Nam, Yong-Tae Kim, Yoo Young Ahn, Nam Hyeong Kim, Ji Hun Kim,Yong Ho Kim, Sungkyunkwan Advanced Institute of Nanotechnology(SAINT), Sungkyunkwan University,Suwon 440-746, South Korea; One-Sun Lee, Qatar Environment and Energy Research Institute, Hamad Bin Khalifa University, Qatar Foundation, P.O. Box 5825, Doha, Qatar

Resume : Supramolecular protein assembly governs most of important biological phenomena such as cell division and enzyme activities and thereby understanding the principle of such interaction gives insights to elucidate the relationship of structure to its function. Self-assembled biomolecules such as DNA and peptide are widely studied for developing new materials as they offer possibilities to integrate biocompatibility into the material applications. Recently, rationally designed self-assembling peptide has been highlighted as a molecular building block because it enables precise modulation of material properties by programming the sequences of peptide that controls the predefined hierarchical structures in a molecular level. Herein we rationally designed a unique nanostructure fiber with self-assembled β-peptide hexameric units. Interestingly, β-peptide nanofibers would allow for associating with single walled carbon nanotube bundles with specific helicities in a manner of superhelical wrapping. We analyzed the structure of β-peptide fibers by small-angle X-ray scattering (SAXS) data that revealed consecutive longitudinal assembly of -peptide hexamers forming nanofiber. The association of -peptide fibers with carbon nanotubes (CNTs) induced extremely stable superhelical protein structure on the surface of CNTs which retained its helical propensity above 100 ℃. The mechanism of superhelical interaction between β-peptide nanofibers and CNTs was fully speculated with MD simulation that calculates the free energy of superhelical wrapping -peptide fibers on CNTs. Furthermore we investigated the electrochemical property of -peptide-CNTs complex through changing the ratio of CNTs to -peptide fibers. The geometrically and electrochemically enhanced superstructure holds a promising future in development of biomaterials applying for biosensors, regenerative medical device, and tissue engineering

Q.P.12
18:00
Authors : Su Yeon Park, Yong-Tae Kim, Yong Ho Kim
Affiliations : SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon 440-746, Korea

Resume : Inducing functionalities on coated surface has been devoted for many years for biomedical uses. Various techniques for immobilizing target function were developed on diverse surfaces. Especially, using biomaterials to control specific functions have been highlighted in biomedical applications. Because this kind of material was advanced in achieving biocompatible and preventing the side effects. However, these materials were only designed for delivering one specific function. Therefore, we suggest a novel strategy to bring multifunctional property synchronically using recombinant proteins. This strategy brought ability to derive two different functions at a time on immobilized bioactive surface. We used mussel based dopamine self-polymerization protein via mussel adhesive protein to tether on the surface and release dual functions by modifying each termination of mussel adhesive protein. The desired functional peptide motif can be recombinant with mussel adhesive protein in E.coli. This recombinant hybrid protein enabled the efficient coating on the surface and more than one functions could be generated for specific targets. As one demonstration, we selectively chose the antimicrobial peptides (AMPs) for one terminal of mussel adhesive protein to enhance the antimicrobial activity. Therefore, the surface was simply functionalized with immobilization of AMPs and successfully examined antimicrobial activity against both gram negative and gram positive bacteria. Therefore, we have collected the list of peptides for specific functions such as wound healing peptide, hormone inducing peptide that can be used in diverse biomedical fields.

Q.P.13
18:00
Authors : Eun Hye Kim1, Jong Min Choi2,3, Jonghyun Kim4, Geonhee Lee5, Won Jin Choi5, Joshep Kwon4, Eun Hee Han4, Seong Hwan Kim1, Young-Ho Chung4,6*, Byung Hwa Jung2,3*, Jeong-O Lee5*
Affiliations : 1 Laboratory of Translational Therapeutics, Pharmacology Research Center, Drug Discovery Division, Korea Research Institute of Chemical Technology, Daejeon 305-600, Republic of Korea; 2 Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea 3 Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 03760, Republic of Korea 4 Division of Bioconvergence Analysis, Korea Basic Science Institute, Daejeon, Korea; 5 Advanced Materials Division, Korea Research Institute of Chemical Technology, Daejeon 305-600, Republic of Korea 6 Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 305-764, Republic of Korea

Resume : We investigate the migration and invasion potential of A549 human lung adenocarcinoma cells grown in micropillar arrays with 2-μm diameter, 16-μm pitch. Previously, it has been shown that micropillar arrays with specific dimensions induce epithelial-to-mesenchymal transition (EMT)-like behavior and morphological changes.1,2 However, EMT-like behaviors induced with micropillar arrays did not exactly coincide with those induced by well-known EMT inducers such as TGF-β. In the present work, we compare the migration potential of A549 cells grown in (2.16) micropillar arrays with those grown in flat substrate, and perform total omics analysis in order to identify cells engineered with micropillar arrays. Cells grown in (2.16) micropillar arrays exhibited distinctively enhanced migratory behavior and invasive potential compared to those grown in flat substrates. Metabolomics analysis of cells unveils highly elevated expressions of all the fatty acid; one of the characteristic of aggressive human cancer cells and primary tumors. Proteomics analysis of cells grown in (2.16) micropillar arrays also show clear increase of proteins related with metastasis of cancer cells such as elongation factor 1 alpha 1 (EEF1A1)3 1. Kim, J. et al. Micropillar arrays as potential drug screens: Inhibition of micropillar-mediated activation of the FAK–Src–paxillin signaling pathway by the CK2 inhibitor CX-4945. Acta Biomater. 27, 13–20 (2015). 2. Badique, F. et al. Directing nuclear deformation on micropillared surfaces by substrate geometry and cytoskeleton organization. Biomaterials 34, 2991–3001 (2013). 3. Tatsuka, M. et al. Elongation factor-1 alpha gene determines susceptibility to transformation. Nature 359, 333–6 (1992).

Q.P.14
18:00
Authors : Brajendra Singh1, Samayendra Kumar2, Bikramjit Basu3, Rajeev Gupta4
Affiliations : 1Centre of Material Sciences, University of Allahabad, Allahabad 211002, India; 2Materials Science Programme, Indian Institute of Technology Kanpur, Kanpur 208016, India; 3Materials Research Centre, Indian Institute of Science, Bangalore 560012, India ; 4Department of Physics, Indian Institute of Technology Kanpur, Kanpur208016, India;

Resume : Bone is a mixture of inorganic calcium phosphate component and organic components. Hydroxyapatite (HAP) and tri calcium phosphate (TCP) are widely recognized as potential bio ceramics for both dental and orthopaedic applications due to their close chemical similarity with the inorganic component of vertebrate bone and tooth mineral. We report the (i) Wet chemical synthesis of doped HAP and TCP (ii) cellular studies (iii) conductivity properties of Silver, Potassium and Magnesium doped HAP and silver doped TCP. Sintered silver and potassium doped HAP and TCP show increase in conductivity while sintered Mg doped hydroxyapatite do not show increase in conductivity. Sintered Sr-HAP (Ca is replace by Sr) does not show the increase in conductivity in presence of Ag doping as observed in HAP. Raman spectra show the characteristics peaks of phosphate, presence of Ag-O bond in doped compositions and absence of O-H bond in TCP. The optical absorption spectroscopic analysis confirms nano and Ag+ ions in sintered HAP and TCP. SEM and TEM investigations confirm the presence of nano and micro sized cubes of silver in doped HAP and TCP. Silver doped HAP compositions exhibit statistically (significant) lower cell viability in comparison to HAp. However, no difference in cellular functionality on doped HAp surfaces, in terms of cell adhesion and proliferation could be qualitatively observed in reference to undoped HAp. In vitro release of Ag+ ions are also quantified using ICP-AES.

Q.P.15
18:00
Authors : Meryem Hatip, Mustafa O Guler
Affiliations : Material Science & Nanotechnology, Bilkent University, Ankara, Turkey

Resume : Supramolecular peptide self-assembly is a powerful bottom-up technique to design and manufacture dynamic complex structures at the nanoscale through non-covalent interactions. Recent advances showed us that the self-assembled peptide nanostructures provide an appealing platform for regenerative medicine, tissue regeneration and stem cell differentiation due to self-supporting and extracellular matrix mimicking properties. Dynamic nature, morphology and supramolecular chirality of the self-assembled peptide nanostructures may affect biological responses. Here, we study various self-assembly conditions and self-assembly mechanism for peptide amphiphile molecules including supramolecular chirality. Change in the environment such as pH or electrostatic interactions, and amino acid sequence of the peptide amphiphiles lead to self-assembled nanostructures with different morphologies. We showed that, type of amino acid residues and trigger forces cause morphological changes. These supramolecular chiral peptide nanostructures were studied in detail by several spectroscopic techniques and imaged by TEM and AFM.

Q.P.16
18:00
Authors : Maria Teresa De Angelis 1, Antonio Paciello 2,3, M. Gabriella Santonicola 1
Affiliations : 1 Department of Chemical Materials and Environmental Engineering, Sapienza University of Rome, 00161 Rome, Italy; 2 Center for Advanced Biomaterials for Healthcare, Istituto Italiano di Tecnologia, 80125 Naples, Italy; 3 Interdisciplinary Research Centre on Biomaterials, University of Naples Federico II, 80125 Naples, Italy

Resume : Interest in stimuli-responsive polymers is steadily increasing especially in the fields of controlled and self-regulated gene delivery. The insertion of appropriate genes into the cells that will repair or replace, is an essential step in gene therapy. Also termed ‘environmental-sensitive’ or ‘smart’, stimuli-responsive polymers experience rapid and reversible changes in their microstructure from a hydrophilic to a hydrophobic state triggered by small changes in the environment, at level of pH, temperature, as well as light, magnetic and electric fields. In this work, we investigate the electrochemical properties of a highly hydrophilic hydrogel based on supramolecular polymers derived from methacrylation of branched polyethyleneimine (PEI-MA). We use electrochemical impedance spectroscopy (EIS) to study the charge transfer at the interface electrode/hydrogel for different applied voltages and buffer solution pH, and we analyze the hydrogel behavior in terms of equivalent circuit models. Results show that the PEI-MA hydrogels respond to both pH and electric fields to an extent that is related to their degree of methacrylation. In addition, we show that DNA can be released in a controlled manner from the hydrogel under the effect of an applied potential, thus demonstrating the possibility of in vitro gene delivery applications for these intelligent hydrogels.

Q.P.17
18:00
Authors : Yawei Sun, Hai Xu, Haiyan Yu, Jiqian Wang
Affiliations : Center for Bioengineering & Biotechnology, China University of Petroleum

Resume : To verify the relationship of structure and property in nucleolipid, a family of amphiphilic nucleolipid with varying hydrophobic chain lengths (from 10 to 16) and various head group (A, C, G and T) were designed and synthesized. Surface tension studies indicated that most of the nucleolipid exhibited defined CMC value and linear relationship between the log CMC or pC20 values with the number of carbon atoms in the alkyl chain, similar to those of conventional anionic surfactants. Besides, the interaction between complementary nucleolipids has been investigated by UV and NMR, revealing that the H-bonding and π-π stacking between nucleobase exhibited little influence in the assembly behavior and surface prosperities of nucleolipid in aqueous solution. Detectable H-bonding could be observed in nucleolipid containing more than 3 nucleobase.

Q.P.18
18:00
Authors : A. De Bonis, M. Curcio, J.V. Rau, M. Fosca, A. Santagata, R. Teghil
Affiliations : Dipartimento di Scienze, Università della Basilicata, Via dell’Ateneo Lucano 10, 85100 Potenza, Italy; Istituto Struttura della Materia – CNR, U.O.S. Potenza, via S. Loja, 85050 Tito Scalo, Italy; Istituto Struttura della Materia – CNR, Via del Fosso del Cavaliere, 100-00133 Rome, Italy

Resume : Since the discovery of the first bioactive glass by Hench, an increasing amount of bioactive glasses have been proposed in order to improve their osteo-conductive behaviour. Recently the incorporation of metallic ions in glass-ceramic materials is considered a new strategy to increase the material bioactivity. Since the biological response is mainly related to the interaction of the device surface with the biological surrounding, these materials are often produced in form of coating of different thickness. Here we report the Pulsed Laser Deposition (PLD) of thin films obtained by ablating a composite target obtained mixing a glass ceramic material of innovative composition (RKKP) [1] with fullerite, in order to improve the coatings mechanical properties. The RKKP-C60 target has been ablated in vacuum by a frequency doubled Nd:YAG laser (t = 7 ns,  = 532 nm, 10 Hz repetition rate), in a wide temperature range, from room up to 600 °C. The characteristics of the deposited coatings have been investigated by microscopical, spectroscopical and diffractometric techniques, namely: Scanning Electron Microscopy, Transmission Electron Microscopy, Atomic Force Microscopy, Fourier Transform Infrared Spectroscopy, micro-Raman spectroscopy, Angular and Energy Dispersive X-ray Diffraction. Vickers microhardness measurements of the composite film–substrate systems have been performed, and the intrinsic hardness of films separated from the composite. The in vitro bioactivity of the obtained films has been studied by soaking sample in Simulated Body Fluid (SBF). [1] A. Krajewski, A. Ravaglioli, A. Tinti, P. Taddei, M. Mazzocchi, R. Martinetti, C. Fagnano, M. Fini, J. Mater. Sci.: Mater. Med. 16 (2005) 119–128.

Q.P.19
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Functionalised and responsive surfaces II : Tommy Nylander
08:30
Authors : Ashutosh Chilkoti
Affiliations : Department of Biomedical Engineering Duke University, Durham, NC 27708, USA

Resume : This talk will discuss the synthesis of a “non-fouling”—protein and cell resistant—polymer brush and its use for the fabrication of a point-of-care clinical diagnostic. In the first part of my talk, I will describe the in situ synthesis of a PEG-like brush polymer, poly(oligo(ethylene glycol) methyl ether methacrylate) (poly(OEGMA)), by surface-initiated atom transfer radical polymerization (ATRP) from different surfaces. I will then describe the use of the POEGMA brush for the fabrication of protein microarrays by inkjet printing antibodies into a macroscopically dry brush on glass. Interestingly, despite the lack of covalent attachment chemistry, this simple methodology results in highly stable and fully bioactive microarrays with femtomolar limits-of-detection in serum and whole blood. Finally I will discuss how this antibody microarray was converted to a point-of-care diagnostic, in which all reagents are printed and stored on the brush.

Q.3.1
09:00
Authors : Katrin Unger, Anna Maria Coclite
Affiliations : Institute of Solid State Physics, Graz University of Technology, Petersgasse 16, 8010 Graz, Austria

Resume : Hydrogels are known for their dynamic swelling response to aqueous environments. Chemical functionalization of the hydrogel surface can be used to add other stimuli-responsive functionalities to the swelling response and thus obtain a smart material that responds to more than one stimulus. Modifying the hydrogel surface with solution-based methods is often problematic due to the damages caused by the permeation of the solvent in the hydrogel. This issue is completely circumvented by the use of solvent-free techniques. A novel multi-responsive hydrogel has been synthetized by initiated chemical vapor deposition (iCVD). The hydrogel incorporates responsiveness to light irradiation and exposure to aqueous environment. The light irradiation modifies the degree of swelling within thin hydrogel film and in turn the mechanical properties. Cross-linked polymers of 2-hydroxyethyl methacrylate (HEMA) were chemically modified by iCVD to introduce azobenzene groups, as confirmed by IR spectroscopy and X-ray photoelectron spectroscopy (XPS). The modified hydrogel swelled in water from 7 to 12 % depending on the percentage of cross-linker and in humidity in the range 2 to 30 %. Through photoisomerization of the azobenzene, the polarity within the hydrogel is modified and as a consequence the affinity to water increases. A clear correlation between light irradiation and swelling properties of the hydrogel will be demonstrated. The materials are all biocompatible and therefore suitable for a great variety of applications, e.g. for cell cultures. Cells respond differently to substrates with different stiffnesses. One extremely innovative use of this material will be that a single substrate can be used to control the cell growth instead of different substrates with different stiffnesses.

Q.3.2
09:15
Authors : Laure Fillaud, Thomas Petenzi, Giorgio Mattana, Vincent Noël, Benoît Piro
Affiliations : Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France

Resume : Molecular detection in liquid environments is an essential step in a wide variety of applications. Electrolyte-Gated Organic Field-Effect Transistors (EGOFETs) have recently attracted considerable attention as liquid-phase sensors because of their low biasing voltages and the intrinsic presence of an electrolyte [1]. In this communication, we propose a novel strategy for the utilisation of EGOFETs as sensors in liquid and physiological media, consisting in the employment of a stimuli-responsive polymer gel (acting as transducing material and electrolyte at the same time), electrochemically deposited on the gate electrode. A pH-sensitive poly(acrylic acid) (PAA) hydrogel was grown on the surface of gold electrodes. These electrodes were characterised at different pH in terms of Electrochemical Impedance Spectroscopy (EIS). Such electrodes were then used as gate electrodes in pBTTT – based EGOFETs which were electrically characterised at different pH. EIS results show an electrochemical capacitance variation in response to pH which is also clearly reflected in the transistors electrical parameters. These preliminary results demonstrate that our devices can be used for the detection of molecular reactions where protons are produced. Moreover, the carboxylic groups contained in PAA chains make these gate electrodes particularly suitable for the functionalisation with bioreceptors able to bind specific target molecules. [1] L. Kergoat et al. Org. Elec. 13 2012, 1-6.

Q.3.3
09:30
Authors : Palange AL1, Key J2, Colasuonno M1, Rizzuti I1, Francardi M1, Di Mascolo D1, Decuzzi P1
Affiliations : 1 Laboratory of Nanotechnology for Precision Medicine, Fondazione Istituto Italiano di Tecnologia (IIT), via Morego 30,16163 Genova, Italy 2 Department of Biomedical Engineering, Yonsei University, Yonseidae-gil, Wonju, Gangwon-do, 220-710, Republic of Korea

Resume : Over the past 10 years, a plethora of nanoparticles for biomedical applications have been proposed exhibiting different sizes - from a few tens to a few hundreds of nanometers; shapes – spherical, cylindrical, discoidal, stellar; and surface properties. Only recently, a few papers have introduced the notion that a fourth parameter – the mechanical stiffness – can also be used for modulating macrophage uptake and blood longevity. Here, 1,000400 nm Discoidal Polymeric Nanoconstructs (DPNs) are engineered with controlled mechanical stiffness. A modified hydrogel-template approach is employed resulting in a precise control of DPN size, shape, surface properties and, most importantly, mechanical stiffness. DPNs are constituted by chains of poly(lactic-co-glycolic acid) (PLGA) and polypropylene glycol (PEG2000) entangled toghether. By changing the paste composition, soft and rigid DPNs (s- and r-DPNs) are synthesized presenting the same geometry and surface charge (-14 mV) but different mechanical stiffness: 1.3 and 15 kPa, respectively. Flow cytometry and in vivo intravital microscopy analyses demonstrate that s-DPNs are less susceptible (~ 4-time) to internalization by macrophages as compared to r-DPNs. Also, PET-CT imaging in mice bearing skin or brain malignancies demonstrate for s-DPNs a ~ 24h circulation half-life and tumor accumulations up to 20% of the injected dose per gram tissue. These unique properties suggest that DPNs can be efficiently used for cancer theranosis.

Q.3.4
09:45
Authors : M. C. Sportelli, E. Tütüncü, R. A. Picca, M. Valentini, A. Valentini, C. Kranz, B. Mizaikoff, N. Cioffi
Affiliations : M.C. Sportelli: Chemistry Department, University of Bari, V. Orabona, 4-70126 Bari, Italy; E. Tütüncü: Institute of Analytical and Bioanalytical Chemistry, Ulm University, Albert Einstein Allee, 11 – 89081 Ulm, Germany; R.A. Picca: Chemistry Department, University of Bari, V. Orabona, 4-70126 Bari, Italy; M. Valentini: Dipartimento Interateneo di Fisica “M. Merlin”, Università degli Studi di Bari “Aldo Moro”, Bari, Italy; A. Valentini: Dipartimento Interateneo di Fisica “M. Merlin”, Università degli Studi di Bari “Aldo Moro”, Bari, Italy; C. Kranz: Institute of Analytical and Bioanalytical Chemistry, Ulm University, Albert Einstein Allee, 11 – 89081 Ulm, Germany; B. Mizaikoff: nstitute of Analytical and Bioanalytical Chemistry, Ulm University, Albert Einstein Allee, 11 – 89081 Ulm, Germany; N. Cioffi: Chemistry Department, University of Bari, V. Orabona, 4-70126 Bari, Italy.

Resume : Surfaces colonization by microorganisms leads to biofilms formation, i.e., bacteria aggregates in which cells are embedded in a self-produced extracellular polymeric matrix (EPS). In this state, bacteria are highly resistant to antimicrobials, thus giving rise to health and environmental problems1. In this perspective, the inhibition of biofilm growth is a crucial issue in the prevention of bacterial infections. Metal/Teflon (Me-CFx) composites deposited via ion beam sputtering (IBS) are well-known antimicrobials2. Specifically, Ag-CFx thin films are considered novel materials of exceptional in-plane morphological and chemical homogeneity. In the present study, Ag-CFx composites were characterized in detail, morphologically and spectroscopically. They were deposited onto IR-inactive regions of a ZnSe attenuated total reflection (ATR) crystal identified following a literature procedure3. This approach allowed monitoring P. fluorescens biofilm growth inhibition induced by the antimicrobial coating. These findings were corroborated by AFM imaging of bacteria incubated on Ag-CFx films, which were deposited onto sterile glass slides. Morphological analyses confirmed that bacterial stress was induced by the composite, leading either to membrane leakage or to bacterial lysis as a function of incubation times. 1 E. Denkhaus et al., Microch. Acta 158 (2007), 1. 2 M.C. Sportelli et al., Sci. Adv. Mat. 6 (2014), 7 and refs. therein. 3 G. T. Dobbs et al., Appl. Spectroscopy 60 (2006), 573.

Q.3.5
10:00
Authors : F. Pappa, V. Karagkiozaki, E. Pavlidou, Th. Choli-Papadopoulou, S. Logothetidis
Affiliations : Nanomedicine Group, Lab for “Thin Films- Nanobiomaterials, Nanosystems & Nanometrology”, Department of Physics, Aristotle University of Thessaloniki, Greece ; Department of Physics, Aristotle University of Thessaloniki, Greece ; Biochemistry Laboratory, Department of Chemistry, Aristotle University of Thessaloniki, Greece

Resume : In Western countries, Neurodegenerative Diseases are a major cause of death and considered to be the pandemic of 21st century. Nanomedicine comes to bear new approaches towards this direction, with great advancements in peripheral nerve reconstruction. Fabrication of suitable scaffolds for neural engineering is critical for the successful nerve regeneration. Electrospinning, a versatile method for nanofiber production has been used to fabricate fibrous scaffolds, with great similarity to the Extracellular Matrix (ECM). To this end, we fabricated conductive nanofiber scaffolds, consisted of biodegradable Polyvinyl alcohol and conductive Poly (3, 4-ethylenedioxythiophene) Polystyrene sulfonate, and proceed towards the evaluation of their surface and mechanical properties, along with degradation rates, emphasizing on the way that manipulate cell growth and adhesion. PC12, a neural cell-line was deposited onto the scaffolds in order to evaluate their cytocompatibility and ability to differentiate into sympathetic neurons in vitro. Biofunctionalization process with biological factors, like peptides and RGD laminins, along with treatment with Nerve Growth Factor took place in order to control and manipulate cell’s differentiation into neurons. MTT assay revealed excellent compatibility along with Confocal microscopy, fact that further reinforced scaffold’s ability to promote neuritis outgrowth. Results indicated that the conductive non-woven scaffolds are represent a unique microenvironment, that can mimic the ECM, promoting cell attachment and proliferation, and via proper surface modification, constitute a promising property that gives impetus to further Nerve Tissue Regeneration applications.

Q.3.6
10:15
Authors : Yujun Feng
Affiliations : Polymer Research Institute, State Key Laboratory of Polymer Materials Engineering, Sichuan University, Chengdu 610065, P. R. China

Resume : A major scientific challenge pertaining to polymers has been to craft smart switchable materials which can reversibly alter their structure and functionality on demand, in response to triggers produced by environmental changes. Among the triggers, CO2 has gained considerable attention owing to its wide availability, biocompatibility and free of contamination. Recently, we have been focusing on developing smart materials whose self-assemlies could be tuned by CO2. • Switchable “living” polymers: these are based on wormlike micelles (WLMs) self-assembled from specifically-tailored surfactants which may be composed of binary surfactant/hydrotrope formulation, or an ultra-long-chain tertiary amine or a polyamine surfactants. Alternating streaming CO2 and N2 (or even air) resulted in the transition between WLMs and spherical micelles, thus the change between high viscoelasticity and low-viscosity macroscopically. • Switchable well-defind triblock and block-random segmented copolymers: the former can form multicompartment micelles (MCMs) with a segregated corona, and can be switched “on” and “off” between MCMs and spherical micelles when sequentially treated with CO2 and N2; while vesicles are formed in the latter case firstly, then fuse hierarchically into giant wormlike micelles; after bubbling CO2 into the copolymer solution up to saturation, the giant worms transform into polymersomes. The vesicles obtained could restore back to wormlike aggregates after depleting CO2 by N2.

Q.3.7
10:30 break    
 
Polymer assemblies : Hai Xu
11:00
Authors : Yun Yan and Jianbin Huang
Affiliations : Peking University

Resume : Cyclodextrins are well-known to form host-guest inclusion complexes with surfactants, whereas it has long been considered the host-guest complexes of surfactant @ CD lose their self-assembling ability due to the coverage of the hydrophobic chain with the hydrophilic outer surface of CD. However, our study shows that in case the hydrophobic chain of surfactant was completely covered by CD, the supramolecular block of host-guest complexes at very high concentration can self-assemble into hierarchical structures driven by the hydrogen bonds between CDs. Vesicles, annular rings, and lamellar structures were observed as the concentration was elevated to 6-10%, 10-20%, and 30-40%,respectively, in the SDS@2β-CD system. Further experiments verified that not only SDS, other surfactants, or even alkanes can also induce the hierarchical self-assembled structures of CD through the formation of channel type CD dimmers or trimmers. It is found that hydrogen bonding is a universal force to drive the supramolecular building block of CD to hierarchical self-assemblies. This opens a new vista for the supramolecular chemistry of cyclodextrins, and is expected to be a facile approach to create novel responsive and functional materials.

Q.4.1
11:30
Authors : Hongyao Yin, Anne-Laure Bulteau, Yujun Feng*, Laurent Billon*
Affiliations : Dr. H. Yin State Key Laboratory of Polymer Materials Engineering, Polymer Research Institute, Sichuan University, Chengdu 610065, PR China. Université de Pau et des Pays de l’Adour (UPPA), Institut des Sciences Analytiques et de Physico-Chimie pour l’Environnement et les Matériaux (IPREM), CNRS UMR 5254, Equipe de Physico-Chimie des Polymères (EPCP), Hélioparc, 2 avenue Angot, 64053 Pau Cedex 9, France Chengdu Institute of Organic Chemistry, Chinese Academy of Sciences, Chengdu 610041, PR China.; Dr. A. L. Bulteau Institut des Sciences Analytiques et de Physico-Chimie pour l’Environnement et lesMatériaux (IPREM), CNRS UMR 5254, Equipe de Chimie Analytique, Bio-Inorganique et Environnement (ECABIE), Hélioparc, 2 avenue Angot, 64053 Pau Cedex 9, France.; Prof. Y. Feng State Key Laboratory of Polymer Materials Engineering, Polymer Research Institute, Sichuan University, Chengdu 610065, PR China.; Prof. L. Billon Université de Pau et des Pays de l’Adour (UPPA), Institut des Sciences Analytiques et de Physico-Chimie pour l’Environnement et les Matériaux (IPREM), CNRS UMR 5254, Equipe de Physico-Chimie des Polymères (EPCP), Hélioparc, 2 avenue Angot, 64053 Pau Cedex 9, France

Resume : Highly ordered porous films have been proven to be excellent candidates as scaffolds for cell culture applications owing to their enlarged surface area and regular pores with narrow size distribution, which are favorable for cell attachment and spreading. In this work, a series of diblock copolymers polystyrene-block-poly(N,N-dimethylaminoethyl methacrylate) (PS-b-PDMAEMA) with different molecule hydrophilicity was synthesized and then used to generate honeycomb-patterned porous film through “Breath Figure” technique. It was found that molecular hydrophilicity plays an essential role in determining film structure, including pore size, porous layers and regularity. Moreover, reversible and tunable surface wettability of these porous films has been achieved simply by alternate introduction and removal of CO2. For the first time, the original hydrophobic porous surfaces are directly used as scaffolds for cell culture with 5% CO2 atmosphere to enhance the interaction with cells during cell culture process, thus resulting in better cell attachment and proliferation, indicative of these smart honeycomb films shows advantages over normal honeycomb films. We anticipate that these unique porous films would not only have wide range of applications in separated hydrophobic or hydrophilic areas, but also can be used to design and develop smart systems where the surface wettability acts as a switch.

Q.4.2
11:45
Authors : Cecilia Masciullo 1, Ilaria Tonazzini 1-2, Marco Cecchini 1
Affiliations : 1 NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Piazza San Silvestro 12,Pisa 56127,Italy; 2 Fondazione Umberto Veronesi, Piazza Velasca 5, Milano 20122, Italy

Resume : Thermal Nanoimprint Lithography (NIL) is a high-throughput and low-cost micro/nanolithography technique that can be applied to a broad range of thermoplastic materials. In fact, by simply applying the right pressure and temperature it is possible to transfer a pattern from a rigid mold surface to the chosen polymer. Usually, high-resolution and large-area NIL molds are difficult to fabricate and very expensive. They are typically made by silicon or other hard materials like nickel or quarts, but after a not very large number of imprinting cycles they starts cracking, becoming unusable. Here, we present the innovative use of perfluoropolyether (PFPE) polymer as material to produce intermediate molds. PFPE intermediate molds allow us to preserve the nanostructured silicon master while transfering micro and nanopatterns to thermoplastic polymers with high fidelity, improving the throughput of the process without affecting the original mold. Several geometries and materials were tested, such as nanometric gratings and nanoripples, and cyclic olefin copolymer (COC) and polyethylene terephthalate (PET). We finally applied this new process to the fabrication of substrates to direct the growth and the differentiation of neuronal and glial cells for possible implementation in devices for peripheral nerve reconstruction after injury.

Q.4.3
12:00
Authors : Marc Jobin, Cédric Pellodi, Cosmin Sandu, Giacomo Benvenuti, Antonin Sandoz, Luc Stoppini
Affiliations : M.J., C.P., L.S. and A.S. ; hepia, University of Applied Sciences of Western Switzerland (HES-SO), 4 rue de la Prairie, CH-1202 Genève, SWITZERLAND . S.C and G.B. : 3D-Oxide, 70 rue G. Eiffel Technoparc, F-01630 St-Genis Pouilly, FRANCE

Resume : We present a cellular test device based on Nb:TiO2 (niobium dooped titania) transparent conductive oxide (TCO) as bio-electrodes. The device is intended to easily provide electrical stimuli to a given cell tissue and to monitor its response. The Nb:TiO2 thin film deposition were performed with combinatorial chemical beam epitaxy (C-CBE) which allows for linear and well controlled gradients of both Nb concentration (ranging from 3% to10%) and oxide thickness in predefined directions (usually orthogonal). We have systematically characterized the Nb:TiO2 films with Atomic Force Microscopy, Scanning White-light Reflectometry, and sheet resistance Finally, we show the device, i.e the electrodes design and patterning in order to perform to electrical stimuli to the various ways to measure the cell’s response.

Q.4.4
12:15
Authors : Yun Yan
Affiliations : Peking University

Resume : Porphyrins are easily aggregate via ?-? stacking in various self-assembled structures. This often resulted in quenching of their fluorescence and the production of singlet oxygen generation. Development of porphyrin self-assemblies that preserves their monomeric photo-physical and chemical properties is highly desired.We developed a series of self-assembled structures of porphyrin which can prevent their ?-? stacking. Porphyrins can be induced to nanotubes, nanoplates, and nanohelixes via out-of-plane coordination or electrostatic interactions, all of them display monomeric photo physical behaviours, which allow retention of sufficient fluorescence and generation of singlet oxygen. Our approach reveals that upon proper strategy of molecular self-assembly, we are able to create ?dispersed? ?-conjugated molecules in their self-assembled structures. We expect this will efficiently preserve the excellent photo-physical and chemical properties, which will provide new options for material science.

Q.4.5
12:30 Lunch    
 
Biomaterials prepared by external stimuli : Jianbin Huang
14:00
Authors : Johannes Bookhold, Thomas Hellweg
Affiliations : Bielefeld University, Department of Chemistry, Physical and Biophysical Chemistry

Resume : Stimuli-responsive surfaces are in the focus of interest for a multitude of applications such as sensors[1], anti-fouling coatings [2] and cell culture substrates [4]. For the latter, coatings made of thermoresponsive Poly-(N-isopropylacrylamide) pNIPAm microgels have been found to allow reversible switching of cell adhesion upon heating and cooling [3-5]. In these works the microgel layer had to be deposited on the substrate intended for use by printing or spin-coating. Hence, the dimensions and material properties of the substrate can strongly influence the adsorption of the microgel particles. The present contribution will review our efforts in this area. Moreover, the preparation of free standing trnasferable membranes from cross-linkable microgels will be presented. Such membranes can be transferred to different surfaces and overcome problems arising from direct deposition of microgels. The approach is based on the deposition of microgels, containing aromatic moieties, by spin-coating the particles on a sacrificial-polyelectrolyte layer. During this work three new monomers, N-(1-Phenylethyl)acrylamide, N-(2,3-dihydro-1H-indene-1-yl)acrylamide and N-Benzhydrylacrylamide, for copolymerization with NIPAm have been synthesized. In a precipitation reaction using a solvent mixture copolymer particles from NIPAm and the aromatic comonomers were obtained, with different comonomer concentration, 2.5 mol-%, 5 mol-% and 10 mol-%. To confirm the incorporation of the aromatic compon[4] S. Schmidt et al., Adv. Func. Mater., 2010, 20, 3235. [5] A. Burmistrova et al. J. Mater. Chem., 2010, 17, 3502.

Q.5.1
14:30
Authors : Frédéric Siffer, Vincent Roucoules, Florence Bally-Le Gall
Affiliations : Institute of Materials Science of Mulhouse (IS2M), CNRS/Université de Haute-Alsace, Mulhouse, France

Resume : Stimuli-responsive materials are able to react to a stimulus provided by their environment and consequently to tailor their properties. Since many phenomena locally occur at the surface of the material, our group work on the design of smart interfaces with switchable properties, which can be elaborated on any kind of material. More specifically, our aim is the elaboration of smart polymer coatings reacting to a temperature change and able to control the immobilization and release of biomolecules at/from a surface. In that context, surface functionalization by plasma polymerization is used to provide these dynamical properties to any material. A thermodynamics study has been carried out to characterize Diels-Alder interfacial reversible reaction on these coatings in comparison with self-assembled monolayers. The reaction progress of cycloaddition was monitored by several surface characterization techniques. It can be noticed that Gibbs free energy is rather similar whatever the accessibility and the density of chemical groups on the substrates are. However, the entropy and enthalpy contributions differ. Plasma polymer thin films seem to be less reactive than monolayers, yet chemically analogous, but the transition-state complex was more ordered. Therefore, the surface characteristics have a strong impact on surface reactivity. Consequently, a fine tuning of surface properties is a key issue to control the immobilization and release of biomolecules via a thermal stimulus.

Q.5.2
14:45
Authors : Abderrahmen Hamdi 1.2,Chohdi Amri 1.2, Rachid Ouertani 1, Hatem Ezzaouia 1
Affiliations : 1. Laboratory of Semi-conductors, Nano-structures and Advanced Technologies, Research and Technology Centre of Energy, Borj-Cedria Science and Technology Park, BP 95, 2050 Hammam-Lif, Tunisia. 2. Faculty of Science of Bizerte, University of Carthage, 7021 Zarzouna, Tunisia.

Resume : Porous silicon powder finds several applications in different areas due to number of properties that make it an attractive material. Within this context, a facile method for the low-cost and large-scale production of Porous silicon (pSi) microparticles, in diverse sizes and shapes, has been used. This method consists of exposing silicon powder to a mixture of HF/HNO3 at specific conditions. Tow samples of the silicon microparticles were analyzed which include the starting metallurgical grade silicon powder and the sample that have been exposed to chemical vapor etching (CVE) . This method lead to the formation of porous silicon nanosponge microparticles. The nanostructured powder has been functionalized with 3-aminopropyl triethoxysilane ( APTES) molecules. These later were intended to work as coupling agent for biosensing applications. Morphologies of pSi microparticles were characterized by scanning electron microscopy (SEM). Fourier transform infra-red (FTIR) and Raman spectroscopic analyses have shown that the APTES molecules are attached to the surface of porous silicon powder.

Q.5.3
15:00
Authors : Caroline Duc, Alexis Vlandas, George G. Malliaras, Vincent Senez
Affiliations : BioMEMS, Univ. Lille, CNRS, ISEN, UMR 8520 - IEMN, F-59000 Lille, France ; BioMEMS, Univ. Lille, CNRS, ISEN, UMR 8520 - IEMN, F-59000 Lille, France ; Department of Bioelectronics, Ecole Nationale Supérieure des Mines CMP-EMSE, MOC, 13541 Gardanne, France ; BioMEMS, Univ. Lille, CNRS, ISEN, UMR 8520 - IEMN, F-59000 Lille, France

Resume : Interfacing biology with electronics promises to give rise to devices that will transform the assessment of a person’s health. Among the different conducting polymers (polypyrroles, polyanilines…) that have been used for this purpose, poly(3,4-ethylenedioxythiophene) doped with poly(styrenesulfonate) (PEDOT:PSS) has emerged as a model material. Despite the fact that applications in bioelectronics require operation in aqueous environment, little is known about the wettability of PEDOT:PSS. By relying on both the often used sessile drop technique and the captive bubble one permitting measurement underwater, we conduct here a detailed investigation of the wetting properties of PEDOT:PSS. Moreover, given the growing use of crosslinking agents to stabilize PEDOT:PSS, we apply these techniques to measure the impact of crosslinker addition on wetting properties and film aging. We show that the crosslinker tends to diminish the impact of the aging, slows down the stabilization (by a factor 2) and increases the hydrophilicity of the film. Finally, we analyze the effect of voltage on the wettability of crosslinked PEDOT:PSS. We observe an advancing contact angle reduction of 30° by applying voltage lower than -1V to the surface. The obtained results provide valuable insights into the actuation properties of PEDOT:PSS film and into its dynamics in water, which should be taken into account in biological environment.

Q.5.4
15:15
Authors : V. Dinca1, L.E. Sima2, A. Bonciu2,3, L. Rusen1, I. Iordache(Urzica)1 and M. Dinescu1
Affiliations : 1National Institute for Lasers, Plasma, and Radiation Physics, Magurele RO-077125, Romania 2Institute of Biochemistry of the Romanian Academy, 296 Splaiul Independentei, 060031,Bucharest, Romania 3University of Bucharest, Faculty of Physics

Resume : The design of topographical and chemical features for engineering smart bio-interfaces with multiple and synergetic functionalities, represent the key point in effective use of hierarchically topographical and chemical bioplatform targeting controlled regulation of stem cell differentiation.Particularly, human mesenchymal stem cells (hMSCs) are of great promise in both basic developmental biology studies and in regenerative medicine, as progenitors of bone cells. Their fate can be affected by various key regulatory factors (e.g soluble growth factors, intrinsic, extrinsic environmental factors) that can be delivered by a fabricated scaffold. For example, when cultured on engineered environments that reproduce the physical features of the bone, hMSCs express tissue specific transcription factors and consequently undergo an osteogenic fate. Therefore, producing smart bio-interfaces with targeted functionalities, represent the key point in effective use of hierarchically topographical and chemical bio-platforms. In this work, we present laser based approaches (e.g. laser texturing) used for design of bio-interfaces aimed at controlling stem cells behavior in vitro. We showed that substrates can be developed to direct hMSCs spatial orientation using laser irradiation to modulate surface microtopography. Various patterns were obtained with progressive smaller inter-trough spacing and depths by decreasing the laser beam overlapping area or using masks systems. The increase in surface area induced increased spreading of cell on all directions, which triggered cell morphology modification towards polygonal shape and consequently increased circularity of stem cell nuclei. Our results demonstrate potential use of laser micropatterned substrates to modulate cell fate during bone implantation.

Q.5.5
15:30 Break    
 
Assemblies of organic and inorganic materials : Junbai Li
16:00
Authors : Lixin Wu
Affiliations : State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, China

Resume : Polyoxometalates (POMs) are known as a class of nanosized metal-oxygen polyanionic clusters with rich chemical composition and diverse structural topology that can lead to interesting applications. To combine the functional features with their dispersivity, porosity, size controllability, and biocompatibility, POMs were used to organize into various media and/or carriers for enhanced functions. But, in general, it is hard to use these clusters in materials and bio-relevant applications directly. An alternative approach is to coassemble them with organic components via forming hybrid supramolecular complex. Considering the anionic feature for all POMs, cationic components are normally chosen for the purpose. Following our previous strategy, herein we would like to report some recent results on the self-assembly and functional properties of hybrid complexes that the organic cations covered on the surface of inorganic clusters through electrostatic interaction (for example in Scheme 1). We designed and synthesized a series of cationic surfactants, linear and dendritic cations bearing oligo(ethylene glycol) monomethyl ether terminal groups, oligopeptide, and used them to combine paramagnetic polyoxometalate K13Gd(β2-SiW11O39)2 and other clusters. The obtained complexes not only constructed interesting self-assemblies, but also performed temperature sensitivity. More significantly, the complexes and their assemblies were found to be potential contrast agent for magnetic resonance imaging. The in-vitro and in-vivo measurements demonstrated the structure and property stability in physiology conditions. References 1.H. L. Chen, Y. Yang, Y. Z. Wang, L. X. Wu, Chem. Eur. J., 2013, 19, 11051–11061. 2.T. Zhang, H.-W. Li, Y. Q. Wu, Y. Z. Wang, L. X. Wu, Chem. Eur. J., 2015, 21, 9028 – 9033.

Q.6.1
16:30
Authors : C. Rodriguez, V. Torres Costa, O. Ahumada, M. Manso Silván
Affiliations : C. Rodriguez; V. Torres Costa; M. Manso Silván Department of Applied Physics, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. O. Ahumada Mecwins, S.L., Parque Científico de Madrid PTM, C/ Santiago Grisolía 2, Tres Cantos, Madrid, Spain

Resume : Biosensing technology is a rapidly advancing field that benefits from the possibility to use the properties of functional advanced materials to analyse biological systems. In particular, nanomechanical systems are very attractive for biological sensing since mechanical interactions are fundamental to biology. Indeed, nanomechanical devices allow measuring forces, displacements and mass changes from cellular processes, and provide high sensitivity and fast responses, which is necessary for the observation of biological processes [1]. On the other hand, among all the functional materials, PSi constitutes an ideal substrate for developing new chemistries owing to its biocompatibility, well-established fabrication methods and large adsorption surface, which allows an enhanced sensitivity [2]. In this work, we will start by reviewing the processes for the PSi formation on microcantilevers and their biofunctionalization in order to trigger its sensitivity as biosensing platform. Secondly, we will describe current approaches based upon modification by self-assembled silane monolayers, which critically depend on the type of process for the activation of PSi. Depending on the molecular structure of the monolayers, the surface presents hydrophobic/hydrophilic properties, allows a molecular selectivity, and a local control of the biomolecular interactions. The surface of the functionalized material will then be biologically activated for the detection of specific genomic or proteomic species applying surface immobilization techniques. Finally, the process of formation of the biorecognition interface will be applied to composite porous silicon-crystalline silicon cantilevers and preliminary sensing results will be shown. [1] M. Calleja, P.M. Kosaka, A. San Paulo, J. Tamayo, Nanoscale. 4, 4925-4938 (2012) [2] S. Stolyarova, S. Cherian, R. Raiteri, J. Zeravik, P. Skladal, Y. Nemirovsky, Sensors and Actuators B, 131, 509-515 (2008)

Q.6.2
16:45
Authors : Gina Kaup, Tom Felbeck, Marina Lezhnina, Mark Staniford, Ulrich Kynast
Affiliations : Fachhochschule Münster University of Applied Sciences Münster, Stegerwaldstr. 39, 48565 Steinfurt, Germany

Resume : In recent investigations we discovered with surprise that nanoscaled phyllosilicates, especially Laponite® RD, exhibit outstanding adsorption properties even for uncharged, non-polar molecules.[1] Laponite® RD (Na0.7(H2O)n{(Li0.3Mg5.5)[Si8O20(OH)4]}) is a commercially available product[2] with unusual properties: for example, clear dispersions in water can readily be obtained due to the morphological and charge characteristics of this nanoclay, and may thus be perceived as a “shuttle” for dyes and even for nanoparticles, eventually enabling its application in the field of smart bioassays. To increase the “playground” of our nanoclays, the rims of the clay platelets can be modified with functional groups, such as amine, thiol or carboxylates, by silanisation of the laponite’s rim Si-OH groups.[3, 4] The attached groups can carry light emitting molecules, plasmonic particles as well as molecules for coupling to organisms and their biofunctions, like sugars or antibodies, for targeted interactions between the clay nanoparticles and biomolecules. This remarkable and widespread scope of laponite properties makes them attractive as smart platforms for new biohybrid materials. References [1] M. M. Lezhnina, T. Grewe, H. Stoehr and U. Kynast, Angew. Chem. Int. Ed., 2012, 51, 10652. [2] http://www.byk.com, 15 January 2016. [3] T. Felbeck, et al., J. Phys. Chem. C, 2015, 119, 12978. [4] G. Kaup, T. Felbeck, M. Staniford and U. Kynast, J. Lumin., 2016, 169, Part B, 581.

Q.6.3
17:00
Authors : Mark C. Staniford1, Marina M. Lezhnina1, M. Gruener2, Christian A. Strassert2 and Ulrich H. Kynast1
Affiliations : 1 Fachhochschule Münster, Univeristy of Applied Sciences, Stegerwaldstraße 39, 48565 Steinfurt 2 Physics Institute and CenTech, Westfälische Wilhelms-Universität Münster, Heisenbergstraße 11, 48149 Münster,

Resume : Phthalocyanines (Pc’s) exhibit unique optical properties, which have raised the interest of numerous scientists. Generally, Pc’s possess very strong absorptions in the deep red spectral range, making them highly interesting for biomedical applications, such as photodynamic therapies of tumours (PDT) and the inactivation of microbial pathogens (PIA), both of which rely on the production of singlet oxygen aided by the Pc [1]. Due to their notorious tendency to agglomerate and insolubility in water, applications of pristine phthalocyanines are hardly realisable, since singlet oxygen generation requires monomeric Pc’s, the aggregation in aqueous ambience persisting even on equipment of the Pc with ionizing substituents. Only recently, we were able to solubilize native MPc’s (M = Al, Cu) with the aid of laponite, a synthetic smectite nanoclay, which furthermore substantially reduced the degree of aggregation and thereby enhanced their fluorescence and singlet oxygen quantum yields. However, for intimate interactions with bacteria, further modifications on the laponite itself were required [2,3]. Next to aqueous solutions of these hybrids, stationary imbeddings into membranes also retained their cytotoxicity, opening a new pathway to antimicrobial surfaces. References 1 M. C. DeRosa, R. J. Crutchley, Coord. Chem. Rev., 2002, 233-234, 351. 2 M. C. Staniford et al, Chem. Commun., 2015,51, 13534. 3 M. Grüner et al, ACS Appl. Mater. Interfaces, 2015, 7 (37), 20965.

Q.6.4
17:15
Authors : Aziliz HERVAULT, Lim MAY, Cyrille BOYER, Alexander DUNN, Toshiaki TANIIKE, Kazuaki MATSUMURA, Derrick MOTT, Shinya MAENOSONO, Nguyen Thi Kim THANH* Email: ntk.thanh@ucl.ac.uk, http:/www.ntk-thanh.co.uk
Affiliations : Aziliz Hervault;1,2,3 May Lim;4 Cyrille Boyer;4 Alexander Dunn;4 Toshiaki Tanikee;3 Kazuaki Matsumura;3 Derrick Mott;3 Shinya Maenosono;3 and N. T. K. Thanh.1,2 1. Department of Physics & Astronomy, University College London, Gower Street, London, WC1E 6BT, UK E-mail: ntk.thanh@ucl.ac.uk; http://www.ntk-thanh.co.uk 2. UCL Healthcare Biomagnetics and Nanomaterials Laboratories, 21 Albemarle Street, London W1S 4BS, UK 3. School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan. 4. School of Chemical Engineering, The University of New South Wales, Sydney, NSW2052, Australia

Resume : Magnetic nanoparticles (MNPs) have emerged as promising technology for their applications in biomedicine. MNPs offer the possibility to deliver drugs and heat at specific locations. The development and characterization of the nanosystem are therefore important steps to ensure that the nanosystem has the desired properties to be used for both magnetic hyperthermia and drug delivery. For example, functionalisation of the MNPs with a suitable polymer layer can provide biocompatibility, colloidal stability, drug loading capability and stimuli-responsive behavior. Magnetic nanosystems that respond to both a change in pH and temperature can give spatial and temporal control over the release of the drug, by making use of the acidic pH found in tumor microenvironment and heat generated from the MNPs when exposed to an alternating magnetic field (AMF) as triggers for the drug release. Moreover, heat has been found to greatly enhance the intracellular drug uptake and the cytotoxic effect of many chemotherapeutic drugs, resulting in a synergistic effect of the therapy. This work aims to develop a dual pH- and thermo-responsive magnetic nanosystem allowing for the triggered release of chemotherapeutic drugs as a consequence of hyperthermia and acidic tumor microenvironment, through breakage of pH and heat labile Schiff base bonds that bind the drug molecules to the polymer. Iron oxide NPs were synthesized by a microwave-assisted co-precipitation method. The thermo-responsive polymer p(DEGMA-co-OEGMA-b-[TMSPMA-co-VBA]), synthesized using RAFT polymerization, was then grafted onto the MNPs surface via a silanisation reaction and the functionalization parameters were optimized. The nanocomposites were characterized by XRD, SQUID, TEM, DLS and TGA. Finally, the heating performances of the nanohybrids were investigated using a magnetic AC hyperthermia system. The MNPs exhibit a superparamagnetic behavior with a saturation magnetization around 75 emu/g. The LCST of the polymer could be easily tuned by varying the initial monomers ratio to be in the hyperthermia temperature range. FTIR analyses confirmed the successful grafting of the polymer by the presence of characteristic carbonyl ester bonds at 1750 cm-1. By varying the MNPs to polymer ratio and the pH of the solution during the functionalization step, suspensions with long term colloidal stability could be obtained. Their potential as magnetic hyperthermia agents was confirmed, demonstrating at the same time the importance of colloidal stability on the heating performances.

Q.6.5
17:30
Authors : A.Vladescu1, M.Badea2, M.Braic1, A.Kiss1, M.Moga2, V.Braic1, E. Posna2, M.Dinu1, M.Balaceanu1
Affiliations : 1National Institute for Optoelectronics, 409 Atomistilor St., Magurele, Romania 2”Transilvania” University of Brasov, 29 Eroilor Blvd., Brasov, Romania

Resume : In dental and orthopaedic implants, the poor implant-bone bonding and infections with both bacteria and funguses are the most serious problems, leading to implantation failure. Therefore, many efforts were directed for finding a solution to obtain a novel implant with high osteoconductive and antibacterial properties, resistant to specific conditions inside the human body. The hydroxyapatite was proposed to be used for coating the metallic implants to increase their osteoconductive ability. However, the low mechanical strength of hydroxyapatite, the relatively low bone bonding rate, high dissolution rate and low antibacterial properties restrict its use in biomedical applications. The aim of the work was to enhance the antibacterial properties of the hydroxyapatite by Ag addition into its structure. The coatings, with different Ag contents, were deposited on Ti based alloy substrates by magnetron sputtering method. The coatings were characterized in terms of elemental and phase composition, morphology, corrosion resistance and antibacterial activity (Staphylococcus aureus MRSA, ATCC 29213, Salmonella Typhimurium ATCC 14028). The influence of the Ag content on the film properties was also analysed. Incorporation of a small amount of Ag into the hydroxyapatite structure resulted in an improved antibacterial efficacy versus Gram-positive bacteria. The best hydroxyapatite with Ag content of 0.7 at. % proved to have the best resistance to the bacteria attack.

Q.6.6
17:45
Authors : Michele Bianchi, Alessandro Russo, Maria Sartori, Annapaola Parrilli, Silvia Panseri, Alessandro Ortolani, Marco Boi, Donald M Salter, Maria Cristina Maltarello, Gianluca Giavaresi, Milena Fini, Valentin Dediu, Anna Tampieri, Maurilio Marcacci
Affiliations : Bianchi; Russo; Sartori; Parrilli; Ortolani; Boi; Maltarello; Giavaresi; Fini; Marcacci: Rizzoli Orthopaedic Institute, via di Barbiano 1/10 Bologna 40136, Italy. Silvia Panseri; Anna Tampieri: Institute for Science and Technology for Ceramics (ISTEC) – National Research Council, via Granarolo 64 Faenza 48018, Italy. Donald M Salter: Institute of Genetics and Molecular Medicine, University of Edinburgh, EH4 2XU Edinburgh, United Kingdom. Valentin Dediu: Institute of Nanostructured Materials (ISMN) – National Research Council, via Gobetti 101, Bologna 40129, Italy.

Resume : Magnetic scaffolds have recently attracted significant attention in tissue engineering, due to the prospect of improving bone formation by acting as a “fixed station” able to accumulate/release targeted growth factors and other soluble mediators in the defect area under the influence of an external magnetic field. Further, magnetic scaffolds are envisaged to improve implant fixation when compared to not-magnetic implants. In a series of experimental studies we investigated the possibility to boost bone regeneration in rabbit femoral condyle defect by implanting hydroxyapatite (HA), polycaprolactone (PCL) and collagen/HA hybrid scaffolds in combination with permanent magnets. The results clearly indicate that the osteoconductive properties of the scaffolds are well preserved despite the presence of a magnetic component. Interestingly, when using bio-resorbable collagen/HA magnetic scaffolds, the reorganization of the magnetized collagen fibers under the effect of the static magnetic field generated by the permanent magnet produces a highly-peculiar bone pattern, with highly-interconnected trabeculae orthogonally oriented with respect to the magnetic field lines. In contrast, only partial defect healing is achieved within the not magnetic control groups. These results open new perspectives on the possibility to improve implant fixation and control the bone morphology of regenerated bone by synergically combining static magnetic fields and magnetized biomaterials.

Q.6.7
18:00
Authors : Vesna Srot1, Ute Salzberger1, Birgit Bussmann1, Julia Deuschle2, Boštjan Pokorny3,4, Ida Jelenko Turinek3 and Peter A. van Aken1
Affiliations : 1. Stuttgart Center for Electron Microscopy, Max Planck Institute for Solid State Research, Stuttgart, Germany. 2. Institute of Materials Science, University of Stuttgart, Stuttgart, Germany. 3. ERICo Velenje, Ecological Research and Industrial Cooperation, Velenje, Slovenia. 4. Environmental Protection College, Velenje, Slovenia.

Resume : Living organisms have capability to form bio-minerals with diverse composition and structure. Many of them are composite materials with excellent physical and mechanical properties [1, 2]. Rodents have opposing long pairs of continuously growing incisors. The front surface of the incisors is enamel; the inner part is softer dentine [3]. The surface of incisors shows characteristic orange-brown color and is identified with the presence of iron [4]. In our study the microstructure and the chemical composition of incisors from the feral coypu (Myocastor coypus Molina) were investigated using transmission electron microscopy (TEM) methods and were combined with the mechanical testing experiments. The layer with variable thickness located at the outer surface of the teeth was detected, possessing a much higher amount of iron compared to the concentration values reported by now. Within the iron-rich surface layer multiple iron containing varieties were identified where iron is detected predominantly in the 3 valence state. With the present discoveries we will deepen understanding of iron incorporation at the nanoscale level and its effect on the microstructural properties. [1] UGK Wegst and MF Ashby, Philos Mag 84 (2004), 2167. [2] AP Jackson and JFV Vincent, J Mater Sci 25 (1990), 3173. [3] BA Niemec in “Small animal dental, oral & maxillofacial disease” (2010), Manson Publishing Ltd, London. [4] EV Pindborg JJ Pindborg and CM Plum, Acta Pharmacol 2 (1946), 294.

Q.6.8
18:15
Authors : Eunsun Kim,a Abdellah Felouat,b Elena Zaborova,b Jean-Charles Ribierre,a JeongWeonWu,a Sébastien Senatore,c Cédric Matthews,c Pierre-François Lenne,c Carole Baffert,d Artak Karapetyan,b,e Michel Giorgi,f Denis Jacquemin,g,h Miguel Ponce-Vargas,i Boris Le Guennic,i Frédéric Fages b and Anthony D’Aléob
Affiliations : a: Department of Physics, CNRS-Ewha International Research Center, Ewha Womans University, Seoul, South Korea b: Aix Marseille Université, CNRS, CINaM UMR 7325, Campus de Luminy, Case913, 13288 Marseille, France c: Aix Marseille Université, CNRS, IBDM UMR 7288,13288Marseille, France d: Aix Marseille Université, CNRS, BIP UMR7281, Marseille, France e: NAS Armenia, Inst Phys Res, Ashtarak2, 0203, Armenia f: Aix-Marseille Université, CNRS, Spectropole FR 1739, 13397 Marseille, France g: Laboratoire CEISAM, UMR CNRS 6230, Université de Nantes, 2 Rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France h: Institut Universitaire de France, 103 Boulevard Saint-Michel, 75005 Paris Cedex 05, France i: Institut des Sciences Chimiques de Rennes, UMR 6226 CNRS-Université de Rennes 1, 263 Avenue du Général Leclerc, 35042 Rennes Cedex, France

Resume : Curcumin is a natural compound that can be used in imaging because it is not toxic and it has even anti-cancerigenic properties. Hemicurcuminoids are based on half of the π-conjugated backbone of curcuminoids. The synthesis of a series of such systems and their borondifluoride complexes are described. The emissive character of those dipolar dyes was attributed to an intraligand charge transfer process, leading to fluorescence emission that is strongly dependent on solvent polarity. Quasi quantitative quenching of fluorescence in high polarity solvent was attributed to photoinduced electron transfer. Those dyes were shown to behave as versatile fluorophores. Indeed, they display efficient two-photon excited fluorescence emission leading to high two-photon brightness values. Furthermore, they form nanoparticles in water whose fluorescence emission quantum yield is less than that of the dye in solution, owing to aggregation-induced fluorescence quenching. When cos7 living cells were exposed to those weakly-emitting nanoparticles, one- and two-photon excited fluorescence spectra showed a strong emission within the cytoplasm that originated from the individual molecules. Dye uptake thus involved a disaggregation mechanism at the cell membrane which restored fluorescence emission. This off-on fluorescence switching allows a selective optical monitoring of those molecules that do enter the cell, which offers improved sensitivity and selectivity of detection in bioimaging purposes.

Q.6.9
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Materials for biomedical devices : Martin Malmsten
08:30
Authors : Carlos Alemán
Affiliations : Departament d’Enginyeria Química, ETSEIB, Universitat Politècnica de Catalunya, Diagonal 647, 08028 Barcelona, Spain

Resume : The development of organic and hybrid surfaces made of synthetic polymers alone or combined with biomolecules for biomedical applications has the potential to revolutionize the treatment of a wide variety of medical conditions. Due to their excellent electrochemical, chemical, and optical properties, conjugated conducting polymers are promising candidates for biomolecule immobilization and subsequent fabrication of nanostructured polymer-based devices for biomedical devices (e.g., biosensors, electrochemical actuators, substrates for tissue engineering, and nanowires). Within this context, I will summarize our most recent findings in the field of polymer-based interfaces for biomedical applications, which involves not only the development of nanostructured polymers and hybrids but also the modulation of their properties through physical treatments (e.g., applying nanoperforation techniques and corona discharge plasma processes).

Q.7.1
09:00
Authors : Sera Shin1, Jaehong Lee1, Jungmok Seo2, Sanggeun Lee1, Dayeong Kim1, Hyuncul Kim1, Soonil Lee1 and Taeyoon Lee1
Affiliations : 1. School of Electrical and Electronic Engineering, Yonsei University, 50 Yonsei-ro, Seodaemoon-Gu, Seoul 03722, Republic of Korea 2. Department of Medicine, Biomaterials Innovation Research Center, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA 02139, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA.

Resume : Surface-enhanced Raman scattering (SERS) have received considerable attention over the past several decades because of their powerful analytic performance for biological and chemical molecular sensing. Conventional SERS substrates which need sample drying procedure have some limitations in disposability of the substrate even using noble metal. To overcome the limitations, SERS substrates combined with closed-channel microfluidic devices have been devoloped for successive detection multiple probing molecules. However, the poor performance of the closed-channel microfluidic SERS substrates due to destruction of metal nanostructures and contaminations from the closed channel should be improved. Here, we describe a novel and facile method to achieve open-channel SERS substrate on droplet guiding track engraved superhydrophobic surface, which can successively detect probing molecules without any contaminations. The SERS-active open-channel microfluidic systems were fabricated by depositing silver dendritic micro/nano structures on a Cu plate which has engraved droplet guiding track and stopper, followed by coating a self-assembled monolayer for the superhydrophobic property. The droplets can be easily rolled-off along the engraved guiding track by the gravitational force without any loss and contamiation and stopped by the droplet stopper at the laser spot for the Raman detection. By using this novel contamination-free SERS platform, successive SERS measurement of droplets were achieved using various target molecules such as rhodamine 6G, nile blue A, and crystal violet with a low concentration of 10-7M.

Q.7.2
09:15
Authors : Josephine Lantoine 1, Thomas Grevesse 1, Laurence Ris 2 and Sylvain Gabriele 1
Affiliations : 1.Mechanobiology & Soft Matter group, Laboratoire Interfaces et Fluides Complexes, Re-search Insitute for Biosciences, CIRMAP, Université de Mons, 20 Place du Parc, B-7000 Mons, Begium ; 2. Département des Neurosciences, Research Insitute for Biosciences, Université de Mons, 20 Place du Parc, B-7000 Mons Belgium

Resume : Understanding brain pathologies requires pertinent in vitro platforms which are even more useful when an architecture evoking the in vivo network organization can be enforced. Futhermore, the effect of the environmental stiffness on the neuronal cells is not well understodd. To address this challenge, we developed robust biomaterials via a microcontact printing technique in order to design in vitro networks of cortical neurons with a controlled architecture. Protein micropatterns allow to impose the geometry of the neuronal assembly by controlling important network features such as the total number of cells, their positions, the number and the length of connections. Our results demonstrate that neuronal networks are formed in vitro through a spontaneous migration process directed by the geometry of laminin micropatterns. Neuronal migration leads to the accumulation of cell bodies on circular islands, which are connected together through axonal elongation directed by thin laminin tracks. First, we combined fluorescence microscopy and pharmacological inhibitors to demonstrate that microtubules play a fundamental role in the neuronal migration process. Then, we used soft hydrogels (E=5 kPa) and intermediate elastomers (E=500 kPa) as surfaces to investigate the influence of the surface rigidity on the network formation. Our results indicate shorter maturation times for neuronal networks grown on soft biosurfaces, suggesting that the motility of cortical neurons is a stiffness-dependent process. Furthermore, we used confocal microscopy and synapsin immunostaining to measure quantitatively the formation of synapses, demonstrating a functional activity of these neuronal networks and the great potential of our biosurfaces .These neuronal networks are therefore reliable smart biointerfaces to test drugs and neuroinflammatory molecules in order to investigate their potential effects on neurons.

Q.7.3
09:30
Authors : Clementine M. Boutry (1), Anaïs Legrand (2), Bob C. Schroeder (1), Paige Fox (2), Zhenan Bao (1)
Affiliations : (1) Departement of Chemical Engineering, Stanford University, Stanford, CA, USA; (2) Plastic & Reconstructive Surgery Stanford University Medical Center, VA Palo Alto, CA, USA

Resume : A new generation of sensors, designed for being implanted in the human body and entirely made of biodegradable materials, is currently emerging. These sensors open the field for new smart and safe diagnostic techniques, without need for a second surgery to remove the devices after their predefined period of use. In the present work, we demonstrate a highly sensitive sensor, entirely made of biodegradable materials, that can measure both strain and pressure. This sensor is designed to monitor the progress of tendon repair after surgery. Classical surgical techniques include sewing the torn ends of the tendon and performing an allograft in case of largely damaged tissues. In addition, new treatments are under investigation at Stanford, where an hydrogel loaded with various bio-species is injected locally in order to stimulate the cellular growth. It is of great interest for surgeons to monitor in situ the recovery after surgery and the healing status of the tissues, with the objective to provide an appropriate care to each patient based on improved diagnosis. The biodegradable sensor, fixed on the tendon, can measure both strain and pressure. The strain is measured with two thin film comb electrodes sliding relative to each other, mounted between two highly flexible elastomer layers. The pressure is measured with a flexible capacitor, where the key element is a microstructured elastic dielectric layer placed between the top and bottom electrode. The combination of strain and pressure sensing capabilities, the biodegradability of the selected materials, the high sensitivity, fast response time and long-term stability of the presented sensor allows its use for tendon tissues recovery monitoring.

Q.7.4
09:45
Authors : Sait Ciftci, Julien Barthes, Philippe Lavalle, Hayriye Özçelik, Christian Debry, Agnes Dupret-Bories, Nihal Engin Vrana
Affiliations : Sait Ciftci a,b#; Julien Barthes a,c#; Philippe Lavalle a,d; Hayriye Özçelik a,d; Christian Debry a,b; Agnes Dupret-Bories a,b; Nihal Engin Vrana a,c a Institut National de la Santé et de la Recherche Médicale, INSERM Unité 1121, 11 Rue Humann, 67000 Strasbourg, France b Hôpitaux Universitaires de Strasbourg, Service Oto-Rhino-Laryngologie, 67098 Strasbourg, France c Protip SAS, 8 Place de l’Hôpital, 67000, Strasbourg, France d Faculté de Chirurgie Dentaire, Université de Strasbourg, 8 rue Sainte Elisabeth, 67000 Strasbourg, France

Resume : The lack of an epithelial layer can significantly decrease the functionality of engineered tissues in respiratory tract, such as engineered trachea. Although incorporation of autologous epithelial cells in the engineered tissue is possible; the necessary amount of cells to cover large surfaces is deterrently high. In order to obviate this problem, we developed a double thin film based epithelial patch that can be applied in-situ for triggering the re-epithelialization. Enzymatically crosslinked gelatin films were first seeded with fibroblasts. Then, a second film layer was incorporated as a support for epithelial cells. In addition, each film layer was loaded with growth factors relevant to the cells they acted as a substrate. By using epithelial model, we have demonstrated the efficacy of growth factor release on cells and the stability of the structure for 7 days. Such patches can be used for fast epithelialization in respiratory tissue engineering while decreasing the necessary number of epithelial cells.

Q.7.5
10:00
Authors : Ayşe Karakeçili, Emre Yüksel
Affiliations : Ankara University Chemical Engineering Department

Resume : Biomaterial-associated infections present a significant threat to patients. The cascade of biofilm formation involves the initial microbial attachment, the formation of microcolonies and proliferation, matrix production and biofilm maturation with propagation of infection. In order to prevent biofilm formation and implant-associated infections several approaches have been reported with the aim of depositing an antimicrobial layer on material surface. Among these strategies, bio-inspired coatings have recently emerged to meet the criteria of low cytotoxicity and long-term stability while minimizing the development of bacterial resistance. Antimicrobial peptides (AMPs) are a vast group of molecules offering several advantages like high efficacy at low concentrations and low propensity for developing bacterial resistance. In this study, Magainin II (Mag II) which is a 23-residue AMP has been immobilized on poly(lactide-co-glycolide) (PLGA) or PLGA/gelatin fibrous membranes prepared by electrospinning. Covalent binding was achieved by carbodiimide/N-hydroxysuccinimide chemistry. The surface immobilization was characterized by X-ray Photoelectron Spectroscopy, Scanning Electron Microscopy and Atomic Force Microscopy studies. The antibacterial activity of the bound Mag II was tested against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Bacterial adhesion tests, SEM and confocal analyses revealed that Mag II played an effective role in reduction of bacterial activity and reduced the number of adhered bacteria more than 50%. AMP immobilization strategy was introduced as a new perspective for modulation of antibacterial properties on PLGA and PLGA/gelatin membranes prepared by electrospinning.

Q.7.6
10:30 Break    
 
Biofunctional supramolecular assemblies : Nigel Slater
11:00
Authors : Xiyun Feng, Jieling Li, Wei Qi, Li Duan and Junbai Li
Affiliations : CAS Key Lab of Colloid, Interface and Chemical Thermodynamics Institute of Chemistry, Chinese Academy of Sciences (CAS) Beijing, China

Resume : Construction of artificial protocells is important to deeply understand complex biological processes and design advanced active biomimetic systems. In living organisms, ATP as the major energy source powers most of the energy consumptions. We will report the reconstitution of ATPase in a lipid bilayer coated microcapsules to fabricate a light or pH triggered ATP generator as well as their mechanism study and application. References 1) Yi Jia, Junbai Li Molecular Assembly of Schiff Base Interactions: Construction and Application Chem. Rev., 115 (2015) 1597-1621. 2) Jieling Li, Yi Jia, Weiguang Dong, Xiyun Feng, Jinbo Fei, Junbai Li Transporting a Tube in a Tube Nano Lett. 14 (2014) 6160-6164. 3) Qiang He, Yue Cui, Junbai Li Molecular Assembly and Application of Biomimetic Microcapsules Chem. Soc. Rev., 38 (2009) 2292-2303 4) Li Duan, Qiang He, Kewei Wang, Xuehai Yan, Yue Cui, Helmuth Möhwald, Junbai Li ATP Biosynthesis Catalyzed by the FoF1-ATP Synthase Assembled in Polymer Microcapsules. Angew. Chem. Int. Ed., 46 (2007) 6996-7000. 5) Wei Qi, Li Duan, Kewei Wang, Xuehai Yan, Yue Cui, Qiang He, Junbai Li ATP Synthesis Catalyzed by Motor Protein CF0F1 Reconstituted in Lipid-coated Hemoglobin Microcapsules. Adv. Mater., 20 (2008) 601-605

Q.8.1
11:30
Authors : Sahar Salehi, Toshinori Fujie, Serge Ostrovidov, Ramin Banan Sadeghian, Ali Khademhosseini
Affiliations : Sahar Salehi WPI-Advanced Institute for Materials Research, Tohoku University, Japan Serge Ostrovidov WPI-Advanced Institute for Materials Research, Tohoku University, Japan Ramin Banan Sadeghian WPI-Advanced Institute for Materials Research, Tohoku University, Japan Toshinori Fujie Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan Ali Khademhosseini WPI-Advanced Institute for Materials Research, Tohoku University, Japan; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, USA; Harvard-MIT Division of Health Sciences and Technology Massachusetts Institute of Technology, Cambridge, MA, USA;Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA; College of Animal Bioscience and Technology, Department of Bioindustrial Technologies, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, Republic of Korea; Department of Physics, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

Resume : Although transplantation of autologous cells has been tested by injection of cell suspensions using a syringe, limited regeneration results were observed due to the low viability of the injected cells, and restricted cell integration into the host tissue. To address these issues, a new approach would be to locally deliver the cells using engineered, cell-loaded substrates. The aim of this study is to develop the flexible nanoribbons from biodegradable polymer that can be aspirated and safely injected into tissues through a conventional syringe needle. In this study, we developed microfabricated poly (lactic-co-glycolic acid) (PLGA) nanoribbons, in order to generate freestanding ribbons loaded with murine skeletal myoblasts (C2C12). Injectability of cell-loaded nanoribbons has been studied using different gages of syringe needles. We observed flexibility of the nanoribbons reduces the mechanical stress on the attached cells. Cell viability, activity, and functionality were not affected after injection of cells adhered to the nanoribbons. In addition cells could safely undergo myogenesis and show the myotubes formation after moving cells to differentiation medium. We anticipate that the injectable nanoribbons promote the formation of muscle tissue and could be a useful tissue engineering technique for local delivery of cells while maintaining the cellular organization, viability, and functionality.

Q.8.2
11:45
Authors : Andreea Belu 1,3, Gokhan Demirel 2, Elmar Neumann 1,3, Dirk Mayer 1,3 and Andreas Offenhaeusser 1,3
Affiliations : 1 Institute of Complex Systems/ Peter Grünberg Institute (ICS-8/PGI-8), Forschungszentrum Jülich, Jülich, Germany; 2 Bio-inspired Materials Research Laboratory (BIMREL), Gazi University, Ankara, Turkey; 3 JARA-Fundamentals of Future Information Technology, Jülich, Germany

Resume : Surface engineering and modification plays a crucial role in biomedical research regarding implants. Since cells are there in contact with artificial solid surfaces, the interface is the part that determines success of a neural implant [1]. Many micro- and nanostructured materials have been employed to provide chemical and physical stimuli to cells. A considerable attention was directed toward 3D, nanostructured polymers. In this work, we explored the role of asymmetric polymeric columnar films on the adhesion and maturation of primary neurons. Poly(chloro-p-xylene) (PPX) can easily be transformed into nanostructured films with high surface area and nanoscale spatial dimension. By means of immunofluorescence staining, SEM and FIB-SEM, we quantified the influence of the nanotextured surface on cell growth in comparison to a planar PPX. The impact of the substrate topography became apparent on the neural development regarding soma size, neuritogenesis, and polarity. We demonstrate that the nanostructures act as geometrical constraints and facilitates an enhancement in neurite branching, axon elongation, and affects growth cone size. Also we observed that the asymmetric orientation of nanofilaments strongly influences the initiation direction of the axon formation, however this directionality is not so pronounced during later stages of axon pathfinding. [1] A. Belu, J. Schnitker, S. Bertazzo, E. Neumann, D. Mayer, A. Offenhäusser, F. Santoro, Journal of Mocroscopy, 2016, accepted

Q.8.3
12:00
Authors : Stefanie Heizmann (1), Antje Kilias (1) (2) (3), Tobias Holzhammer (1), Patrick Ruther (1), Ulrich Egert (1) (2), Maria Asplund (1)
Affiliations : (1) Department of Microsystems Engineering (IMTEK), University Freiburg, Germany; (2) Bernstein Center Freiburg, University Freiburg, Germany; (3) Faculty of Biology, University Freiburg, Germany

Resume : Neuronal signal recordings in vivo, and subsequent histological analysis, are essential aspects for investigating neuronal disorders like epilepsy in animal models. We show that penetrating microelectrodes used for long-term signal recording can be combined with a neurotracer delivery coating greatly facilitating the retroactive tracing of the recorded neurons. An electro-active electrode coating, PEDOT/PSS/neurotracer, applied on the recording sites allows on-demand delivery of dye by defined electrical stimuli. This confines the release to the experimental end point, allows precise labelling of neurons in direct vicinity of the recording site and avoids dye interference with neural function during recordings. Positively charged dye could be electrostatically soaked into the polymer matrix. The unspecific dye adsorption to the surface of the neural probe was successfully prevented by introducing a sacrificial Parylene C layer. This layer is peeled off the electrode chip after coating the electrodes with PEDOT/PSS incorporating the neurotracer dye. The strong cohesion of the Parylene film supports a residual free mechanical peeling off the surface leaving a perfectly clean passive surface underneath. This coating concept allows a contamination free fabrication of dye delivery systems, can easily be combined with a PEDOT-based on-demand dye release which ultimately enhances the precision of neuronal tracing in tissue.

Q.8.4
12:15
Authors : Jing Zhang, Jeff Penny, Jian R Lu
Affiliations : Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester

Resume : Because the small intestine plays a crucial role in absorbing nutrients and drugs, research into understanding its working mechanisms has been expanding over last few decades. Various models have been developed to better investigate absorption and metabolism, among which the Caco-2 cell model is the most widely used. However, this monolayer cell model may lead to inaccurate experimental results and faulty conclusions due to its over-simplification. In our present study, an innovative three-dimensional cell model which simulates in vivo small intestine epithelium, subepithelial fibroblast network and extracellular matrix was developed to enhance the relevance of the drug absorption research. In comparison with the Caco-2 monolayer cell model, the 3D model shows reduced transepithelial electrical resistance (TEER) and higher levels of alkaline phosphatase (ALP) activity in microvilli. Two main efflux transporters – P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) were also studied, and the decreased P-gp activity and increased BCRP activity indicated a better simulation to the in vivo intestine functioning. In conclusion, our reconstructed 3D cell model is better than the widely used Caco-2 monolayer cell model and much closer to the human small intestine at the structural, physiological and functional levels.

Q.8.5
12:30 Lunch    
 
Interactions with Lipid membranes 1 : John Webster
14:00
Authors : Gregory Hardy, Gene Wong, Joseph Shapter, Munir Alam, and Stefan Zauscher
Affiliations : Duke University

Resume : Although the neutralizing mechanisms of two,rare neutralizing antibodies (NAbs), 2F5 and 4E10, which bind to membrane-proximal external region (MPER) of viral gp41, represents a promising framework for the design of new HIV-1 liposomal vaccine candidates, this mechanism remains poorly understood. It is known that 2F5 and 4E10 are required to first associate with HIV-1 lipids before binding to the target MPER antigen, however, little is known about how lipid membranes contribute to NAb-antigen binding. To this end we focused on recreating the lipid phase organization (i.e., domain formation) of native membranes by using supported lipid bilayers (SLBs). To recreate the HIV-1 envelope, we used amphipathic, α-helical peptides as a catalyst to generate complex SLBs that have a high cholesterol content and contain multiple lipid types, and contain a liquid-disorderd (Ld) and liquid-ordered (Lo) phase. We used atomic force microscopy (AFM) to visualize membrane domains, antigen presentation, and antibody-membrane interactions on our SLB surfaces. Our experiments on complex SLBs demonstrate that 2F5/4E10 do not interact with the highly ordered gel and Lo domains in the SLB but exclusively bind to the Ld phase. This suggests that 2F5/4E10 require low membrane order and weak lateral lipid-lipid interactions to insert into the hydrophobic membrane interior. Thus, vaccine liposomes that primarily contain an Ld phase are more likely to elicit the production of lipid reactive, 2F5- and 4E10-like antibodies, compared to liposomes that contain an Lo or gel phase. Furthermore, we show that the presence of the MPER656 antigen can severely limit the Ld area available for antibody interactions. Subsequently, this reduces the amount of MPER656 that is accessible for 2F5/4E10 binding. If Ld forming lipid components are used in vaccine liposomes, it is thus important to ensure that the presence of antigen does not inhibit large-scale Ld formation.

Q.9.1
14:30
Authors : Lina Nyström
Affiliations : Department of Pharmacy, Uppsala University, P.O. Box 580, SE-752 32 Uppsala, Sweden

Resume : Microgels are weakly cross-linked polymer colloids, which can be designed to display responsive volume transitions triggered by a range of parameters. In the context of drug delivery, microgels are of particular interest as carriers for biomacromolecular drugs, such as peptides and proteins, since they offer a water-rich environment for incorporated macromolecular drugs, thus reducing detrimental conformational changes and aggregation within the delivery system. Furthermore, microgels offer various additional benefits as delivery systems for such drugs, including protection against enzymatic degradation and controlled or triggered release. While microgel suspensions and their use as delivery systems are becoming increasingly understood, much less is known about surface-bound microgels as carriers of biofunctional molecules. Addressing this, we here report on work done to elucidate factors determining volume transitions of electrostatically triggered surface bound microgels, as well as their use as delivery systems for peptides. In doing so, we investigate effects of microgel charge density, pH, and ionic strength on microgel volume transitions at surfaces, surface-induced microgel deformation and nanomechanical properties, as well as consequences thereof for peptide loading and release, using a battery of experimental techniques, including AFM PeakForce QNM, QCM-D, ellipsometry, and confocal and cryoTEM microscopy.

Q.9.2
15:00
Authors : Daniela Ciumac, Zongyi Li, Peixun Li, Richard Campbell, Arwel Hughes, Luke Clifton, John Webster, Jian R Lu
Affiliations : Biological Physics Laboratory, School of Physics and Astronomy, University of Manchester Institute Laue Langevin ISIS Neutron Facility, STFC

Resume : A series of short amphiphilic peptides has been shown to kill bacteria whilst remaining benign to mammalian host cells In co-culturing. To help understand the molecular basis underlying efficacy and selective responses, we have undertaken careful surface pressure measurements of the binding of lead peptides to different lipid monolayer models. Neutron reflection in combination with deuterium labelling has also been used to help determine the amount and location of the peptide bound or associated with different model membranes. This talk will illustrate the main factors that affect peptide selective binding and association to different lipid membranes. The knowledge helps the structural design of the peptides leading to useful insight in the improvement of their selectivity and efficacy.

Q.9.3
15:15
Authors : E. Gatto (a), R. Lettieri (a), A. Colella (a), F. Leonelli (b), L. Stella (a), M. Venanzi (a)
Affiliations : (a) Dipartimento di Scienze e Tecnologie Chimiche, Università degli Studi di Roma Tor Vergata, Roma, Italy; (b) Dipartimento di Biologia Ambientale, Università degli Studi di Roma La Sapienza, Roma, Italy

Resume : Supported lipid membranes represent an elegant way to design smart fluid biointerfaces able to mimic the physico-chemical properties of biological membranes, providing an excellent model system for studying the surface chemistry of the cell. Furthermore, supported lipid membranes are accessible to a wide variety of surface-specific analytical techniques, providing smart biointerfaces for biotechnological applications [1], such as the design of chemical and biomedical sensors. In this contribution we describe a new lipid-based sensor for the detection of the thymidine phosphorylase (TP) enzyme, one of the most known biological markers of solid tumors. This enzyme promotes tumor growth and metastasis and is overexpressed in the presence of cancers, so that also its blood levels increase [2]. To achieve this goal, one of the most used anticancer drugs, i.e. the pyrimidine analogue 5-fluorouracil (5-FU) [3] has been properly functionalized with a C-12 aliphatic chain in order to be inserted into gold supported lipid membranes. The interaction of TP with the 5-FU inhibitor and its derivatives has been firstly evaluated in solution by fluorescence measurements, then the derivatives have been inserted into a lipid bilayer linked to a gold surface. The supported lipid biointerfaces have been characterized by ellipsometry, AFM and electrochemical techniques. The TP interaction with the substrate has been quantitatively evaluated by quartz crystal microbalance, following the oscillation frequency of the QCM crystal, making this system a very promising sensor for the detection of TP concentration in blood. [1]. Castellana, E. T.; Creme, P. S. Solid supported lipid bilayers: From biophysical studies to sensor design. Surface Science Reports 61, 429–444 (2006). [2]. Haraguchi, M.; Komota, K.; Akashi, A.; Furui, J.; Kanematsu, T. Occurrence of hematogenous metastasis and serum levels of thymidine phosphorylase in colorectal cancer. Oncol. Rep. 10, 1207-1212 (2003). [3]. Malet-Martino, M.; Martino, R. Clinical studies of three oral prodrugs of 5-fluorouracil (capecitabine, UFT, S-1): a review. The Oncologist 7, 288-323 (2002).

Q.9.4
15:30
Authors : Thomas Werzer1, Oliver Bixner2, Günter Trettenhahn1, Eva-Kathrin Sinner2 Wolfgang Kautek1
Affiliations : 1 University of Vienna, Department of Physical Chemistry, Vienna, Austria 2 University of Natural Resources and Life Sciences (BOKU), Institute of Synthetic Bioarchitectures, Vienna, Austria

Resume : Recently, polymersomes have proven to be interesting model systems for studying polymeric membranes1, in vivo tumour-shrinkage2 and drug delivery vehicles3. Polymersomes are composed of amphiphilic block copolymers, that is, they consist of a hydrophilic or polar part and a hydrophobic or apolar part. Some of their advan-tages are their ability of encapsulating drugs and molecules, as well as a low leakage rate, which can be tuned by adjustment of the hydrophobic block size. This makes them ideal model systems for investigations on the carrying and release activities of drugs and dyes in the human body.4 Nevertheless, information of the interaction of polymersomes with proteins is not widely explored. Polymersomes are also capable of forming 2-D lipid bilayers on solid supports, therefore making them attractive for the application as novel biosensors.5 In this study, the adsorption behaviour of different polymersomes (charged and neu-tral) and liposomes on gold has been studied in phosphate buffered saline solution with the Electrochemical Quartz Microbalance (EQMB; or Quartz Crystal Microbal-ance, EQCM). Moreover, the influence of Bovine Serum Albumin on the stability of the adsorbed vesicles has been investigated. 1. D. Wu, M. Spulber, F. Itel, M. Chami, T. Pfohl, C. G. Palivan and W. Meier, Macromolecules, 2014, 47, 5060-5069. 2. F. Ahmed, R. I. Pakunlu, G. Srinivas, A. Brannan, F. Bates, M. L. Klein, T. Minko and D. E. Discher, Molecular Pharmaceutics, 2006, 3, 340-350. 3. E. Amstad and E. Reimhult, Nanomedicine, 2012, 7, 145-164. 4. K. Sato, E. Abe, M. Takahashi and J. I. Anzai, Journal of Colloid and Interface Science, 2014, 432, 92-97. 5. S. May, M. Andreasson-Ochsner, Z. Fu, Y. X. Low, D. Tan, H.-P. M. deHoog, S. Ritz, M. Nallani and E.-K. Sinner, Angewandte Chemie, 2013, 125, 777-781.

Q.9.5
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Interactions with lipid membranes 2 : Giovanni Marletta
08:30
Authors : Luke Clifton, Maximilian Skoda, Stephen Holt, Anton le Brun, Arwel Hughes, Jeremy Lakey
Affiliations : ISIS Pulsed Neutron and Muon source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxford; ISIS Pulsed Neutron and Muon source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxford; Bragg Institute, Australian Nuclear Science and Technology Organisation, Locked Bag 2001, Kirrawee DC, NSW 2232, Australia; Bragg Institute, Australian Nuclear Science and Technology Organisation, Locked Bag 2001, Kirrawee DC, NSW 2232, Australia; ISIS Pulsed Neutron and Muon source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxford; Institute for Cell and Molecular Biosciences, Newcastle University, Framlington Place, Newcastle upon Tyne, NE2 4HH, United Kingdom

Resume : Bacteria are differentiated into two main groups, Gram-positive or Gram-negative, based on the Gram stain which detects the thick peptidoglycan cell wall of gram positive bacteria. Gram-negative bacteria are of particular biomedical and technological interest due to their role in disease, the increasing antibiotic resistance of some species and their utility in many biotechnological processes. The Gram-negative bacterial outer membrane (GNB-OM) is asymmetric in its lipid composition with a phospholipid-rich inner leaflet and an outer leaflet predominantly composed of lipopolysaccharides (LPS). LPS is a polyanionic molecule, with numerous phosphate groups present in the Lipid A and core oligosaccharide regions. Using a series of systems1-4 we have attempted to create GNB-OM assays which are amenable to molecular level characterisation. These systems are always planar samples deposited at either air/liquid or solid/liquid interfaces and range from simple anionic phospholipid monolayers to asymmetrical GNB-OM models which float ~2 nm above a solid/liquid interface. These membrane models have provided new insights into the OM and interactions with it; from providing conformation of the activity of anti-bacterial proteins and the role of the lipopolysaccharide polysaccharide chains in blocking antimicrobial protein binding to describing the importance of divalent cations in stabilising the OM. References 1. Clifton, L. A., Skoda, M. W. A., Daulton, E. L., Hughes, A. V, Brun, A. P. Le, Lakey, J. H., Holt, S. A., and Hughes, V. (2013) Gram-negative bacterial outer membrane mimic Asymmetric phospholipid : lipopolysaccharide bilayers ; a Gram-negative bacterial outer membrane mimic. J. R. Soc. Interface 10, 20130810 2. Clifton, L. A., Holt, S. A., Hughes, A. V., Daulton, E. L., Arunmanee, W., Heinrich, F., Khalid, S., Jefferies, D., Charlton, T. R., Webster, J. R. P., Kinane, C. J., and Lakey, J. H. (2015) An Accurate In Vitro Model of the E. coli Envelope. Angew. Chemie Int. Ed. 54, 11952–11955 3. Clifton, L. A., Skoda, M. W. A., Le Brun, A. P., Ciesielski, F., Kuzmenko, I., Holt, S. A., and Lakey, J. H. (2015) The Effect of Divalent Cation Removal on the Structure of Gram-negative Bacterial Outer Membrane Models. Langmuir 31, 404–412 4. Le Brun, A. P., Clifton, L. A., Halbert, C. E., Lin, B., Meron, M., Holden, P. J., Lakey, J. H., and Holt, S. A. (2013) Structural Characterization of a Model Gram-negative Bacterial Surface Using Lipopolysaccharides from Rough Strains of Escherichia coli. Biomacromolecules 14, 2014–2022

Q.10.1
09:00
Authors : Houcem Maaoui, Roxana Jijie, Radouane Chtourou, Sabine Szunerits and Rabah Boukherroub*
Affiliations : biomaterials

Resume : Complications related to infectious diseases have significantly reduced, particularly in the developed countries, due to the availability and use of a wide variety of antibiotics and antimicrobial agents. However, excessive use of antibiotics and antimicrobial agents increased the number of drug resistant pathogens, and this has resulted in a significant threat to public health. The inexorable rise in the incidence of antibiotic resistance in bacterial pathogens, coupled with the low rate of emergence of new clinically useful antibiotics, has refocused attention on finding alternatives to overcome antimicrobial resistance. Novel strategies aiming to reduce the amount of antibiotics, but able to prevent and treat animal and human infections should be investigated, evidenced and approved. Among the various approaches, the use of nanotechnology (engineered nanoparticles) is currently the most promising strategy to overcome microbial drug resistance. Due to their small size, nanoparticles can surmount existing drug resistance mechanisms, including decreased uptake and increased efflux of drug from the microbial cell, biofilm formation, and intracellular bacteria. Herein, we propose to use Prussian blue nanoparticles (PB NPs) as efficient photothermal agents under NIR (810 or 980 nm) irradiation for efficient ablation of virulent and antibiotic resistant pathogens, including virulent strains of E. coli associated with urinary tract infection, and methicillin-resistant S. aureus. Although, PB NPs have been investigated for photothermal ablation of cancer cells in the wavelength range of 700-800 nm, to the best of our knowledge there has been no report on the use of these nanoparticles for bacteria treatment. Moreover, the use of 980 nm wavelength has never been described before. Our results clearly show that PB NPs are very effective killing of Gram positive and Gram negative bacteria under 810 or 980 nm irradiation in a concentration-dependent manner. Moreover, under 980 nm irradiation mammalian Hela cells exhibited minimal toxicity up to a PB NPs concentration of 100 µg mL-1, while at this concentration a 100% bacteria killing was achieved. This interesting finding suggest thats these nanoparticles could be potentially used for selective targeting of bacteria over mammalian cells.

Q.10.2
09:15
Authors : Costanza Montis, Alejandro Marín Menendez, Chiara Magnani, Teresa Diaz Calvo, Pierre Joseph, Kostas Hatzixanthis, Christopher Morris, Michael McArthur, Debora Berti
Affiliations : University of Florence and CSGI, Department of Chemistry “Ugo Schiff”, 50019, Sesto F.no, Florence, ITALY; Procarta Biosystems ltd, Norwich Bioincubator, Norwich NR4 7TJ, UK; University of Florence and CSGI, Department of Chemistry “Ugo Schiff”, 50019, Sesto F.no, Florence, ITALY; School of Medicine University of East Anglia, Norwich NR4 7TJ, UK; LAAS – CNRS, Nano Engineering and Systems Integration, BP 54200 31031 Toulouse cedex 4, FRANCE; School of Pharmacy
 University of East Anglia, Norwich NR4 7TJ, UK; School of Pharmacy 
University of East Anglia, Norwich NR4 7TJ, UK; School of Medicine
 University of East Anglia, Norwich NR4 7UQ, UK; University of Florence and CSGI, Department of Chemistry “Ugo Schiff”, 50019, Sesto F.no, Florence, ITALY

Resume : Bacterial resistance to antimicrobials is a global threat that requires development of innovative therapeutics that circumvent its development. We designed a novel nanostructured antibiotic, composed of a bolaamphiphile (12-bis-THA) and an oligonucleotide (transcription factor decoy, TFD). The hypothetical mechanism of action of 12-bis-THA/TFD consists of two steps: 1) the positively charged assemblies destabilize bacterial membranes; 2) once internalized and released the TFD interferes with RNA transcription, inhibiting essential genes required for growth or disease progression. To get mechanistic insights of specific targeting toward bacterial membranes, we studied the interaction of 12-bis-THA/TFD with model membranes of different composition and curvature. Fluorescence and Dynamic Light Scattering on liposomes revealed the specific role of a typical bacterial lipid, cardiolipin, in the destabilization of the membrane and release of the TFD from the assembly. Confocal Microscopy coupled with Microfluidics and Fluorescence Correlation Spectroscopy (FCS) on Giant Unilamellar Vesicles (GUVs) highlighted the role of lipopolysaccharides in determining the interaction and penetration of the TFD inside the GUVs. Finally, Confocal Microscopy and FCS data on E. coli, Gram-negative prototypical bacteria, confirmed the findings on membrane models, validating our synthetic model and allowing to hypothesize an interaction pathway of the nanostructured antibiotic with bacteria.

Q.10.3
09:30
Authors : Dr Ian M Tucker, Dr M Wade, Prof T Wess
Affiliations : Dr Ian M Tucker, Unilever R&D Port Sunlight Dr M Wade, School of Optometry, University of Cardiff Prof T Wess, Executive Dean, Charles Sturt University, Wagga Wagga, New South Wales, Australia

Resume : Human hair exhibits obvious macroscopic variations, in terms of colour, structure, form and physical strength. This is most obvious between individuals of different ethnic backgrounds. The aim of this work was to investigate the origins of the structural variations between the nanoscale, supramolecular and or sub-molecular distance scales, using high resolution X-ray microfocus studies allied with PCR. Results indicate a structural correlation in terms of amount of lipid and orientational ordering in the different hair types.

Q.10.4
10:00
Authors : Nigel Slater
Affiliations : Department of Chemical Engineering and Nanotechnology, University of Cambridge

Resume : In this talk, I will first introduce the historic development of the vectors that can mediate cell membranes and aid the internalisation of proteins, DNA and siRNA, followed by describing recent advances in the design of synthetic polymers and peptides as new vectors, with several examples coming out of my own research group. I will end my talk by pointing to some of the major technical challenges that still constrain the practical application of this attractive technology.

Q.10.5
10:30 Break    
 
Lipid and polypeptide based nanostructures and membrane lytic interactions : Jian R Lu
11:00
Authors : Debora Berti
Affiliations : CSGI and University of Florence, Department of Chemistry “Ugo Schiff”, 50019, Sesto F.no, Firenze, Italy.

Resume : Biomembrane activity of inorganic or polymeric nanoparticles (NP), defined as their tendency to structurally modify and/or permeate in biomembranes, requires full elucidation to optimize biomedical properties or the minimization of health risks in consumer products. To address these interactions in two case studies, we used different model membrane systems, giant unilamellar vesicles (GUVs), supported lipid bilayers (SLB) and liposomes, challenged with: a) A nanostructured antibiotic drug, composed of a bolaamphiphile and a transcription factor decoy, TFD, that self-assemble in solution to form NP (70 nm). The investigation with model bacterial membranes provided a sketch for a hypothetical mechanism of action: first they destabilize bacterial membranes and then, once internalized, the TFD is released and, in bacteria, it interferes with RNA transcription, inhibiting sporulation. b) Gold NPs, of different size, shape and surface coating. Their effect on bilayer morphology, permeability and fluidity present strong differences or similarities, depending on the length scale, from the colloidal to the molecular domains. After a surface-energy driven adsorption, the NPs stiffen the region of contact and “freeze” the lipids in raft-like nanoscale domains. In vitro experiments performed on E. Coli (a) and rat macrophages (b) challenged with the same NPs, indicate a close analogy with the observations in mimetic models, providing validation of our experimental approach.

Q.11.1
11:30
Authors : Rongjun Chen
Affiliations : Department of Chemical Engineering, Imperial College London, South Kensington Campus, London, SW7 2AZ, United Kingdom

Resume : It remains a big challenge to effectively deliver molecules, in particular macromolecules, through membrane barriers. There is a need to better understand the mechanisms of entry into the cell cytoplasm and nucleus in order to design optimal delivery systems for agents of pharmaceutical interest. This presentation will cover our recent efforts to design, synthesis and in- vitro/in-vivo evaluation of novel bio-functional polymers. The pH-responsive, metabolite-derived polymers are designed to mimic factors that enable efficient viral transfection. Strict control over the size, structure, hydrophobicity- hydrophilicity balance and aromaticity of the polymers can effectively manipulate their conformational characteristics and dynamic behaviour in aqueous solution and their interactions with lipid membrane, cell and tissue models. It has been demonstrated that the biomimetic polymers can traverse the extracellular matrix in 3-D multicellular spheroids, reach individual cells in the tumour models, and enable efficient cellular entry of therapeutic payloads (e.g. siRNA and therapeutic proteins) and imaging moieties into the cytoplasm or the nucleus. This could represent a promising therapeutic delivery platform, suitable for research and theranostic applications in the treatment of various diseases including cancer.

Q.11.2
12:00
Authors : Aleksandra P. Dabkowska, Christopher Hirst, Meina Wang, Maria Vadeperas, Gunnar K. Pálsson, Luke Clifton, Justas Barauskas, Sofi Nöjd, Sebastian Lages, Nina-Juliane Steinke, Björgvin Hjörvarsson, Tommy Nylander,
Affiliations : Division of Physical Chemistry, Lund University, P.O. Box 124, SE-22100 Lund, Sweden and NanoLund, Lund University, P.O. Box 118, SE-22100 Lund, Sweden; Institut Laue Langevin, France and Department of Physics, Uppsala University, Box 530, S-751 21 Uppsala, Sweden; ISIS Pulsed Neutron and Muon Source, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell Oxford Campus, Didcot, Oxfordshire, OX11 OQX, UK; Camurus AB, Ideon Science Park, Gamma Building, Sölvegatan 41, SE-22379 Lund, Sweden and Biomedical Science, Faculty of Health and Society, Malmö University, SE-20506, Malmö, Sweden; Division of Physical Chemistry, Lund University, P.O. Box 124, SE-22100 Lund, Sweden; Max IV Laboratory, Lund University, P.O. Box 118, SE-22100 Lund, Sweden; ISIS Pulsed Neutron and Muon Source, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell Oxford Campus, Didcot, Oxfordshire, OX11 OQX, UK; Department of Physics, Uppsala University, Box 530, S-751 21 Uppsala, Sweden;Division of Physical Chemistry, Lund University, P.O. Box 124, SE-22100 Lund, Sweden

Resume : We show that polymer microgels with a diameter of about 55 nm can be embedded within films of non-lamellar lipid liquid crystalline films to form responsive nanostructured surfaces. A simple spin-coating procedure is developed to form these hybrid films from the solubilized components followed by equilibration in excess water. The mixed lipid layers, which are composed of glycerol monooleate and diglycerol monooleate lipids with poly(Nisopropylacrylamide)(PNIPAM) microgels, form hybrid films with reverse bicontinuous cubic phase structure that are capable of responding to temperature stimulus. The thickness, hydration and internal structure of the films are characterized by spectroscopic ellipsometry, attenuated total reflectance FTIR, neutron reflectivity and small angle X-ray scattering. We demonstrate that the microgel particles act as thermoresponsive controllers for the hydration of the lipid films. When the temperature is increased to reach the volume phase transition point of the PNIPAM microgels, i.e. the particles change from the swollen to the collapsed state, water is release from the surface film while the lipid matrix remains intact. The observed surface structure and control of layer hydration can be used to manipulate properties of non-lamellar lipid liquid crystalline layers. These new nano-materials based on lipid-polymer-based responsive layers opens up for new sensor applications where temperature triggered control of the layer hydration are required.

Q.11.3

No abstract for this day


Symposium organizers
Giovanni MARLETTAUniversity of Catania

Laboratory for Molecular Surfaces and Nanotechnology V.le A.Doria 6 Catania Italy

+39 95 7385130
gmarletta@unict.it
Hai XUChina University of Petroleum

Centre for Bioengineering and Biotechnology 66 Changjiang West Road Huangdao Economic Development Zone Qingdao P.R. China

+86 532 86981318
xuh@upc.edu.cn
Jian R. LUUniversity of Manchester

Biological Physics Lab, School of Physics and Astronomy Room 3.14, Schuster Building Oxford Road Manchester M13 9PL UK

+44 161 2003926
j.lu@manchester.ac.uk
Nigel SLATERUniversity of Cambridge

Department of Chemical Engineering and Biotechnology Pembroke Street Cambridge CB2 3RA U.K.

+44 1223 762953
nkhs2@cam.ac.uk
Tommy NYLANDERLund University

Physical Chemistry, Department of Chemistry PO Box 124 SE-221 00 Lund Sweden

+46 46 2228158
Tommy.Nylander@fkem1.lu.se