Bioinspired and Biointegrated Materials as Frontiers Nanomaterials IV

Resume : Organic chemistry offers tremendous opportunities for the synthesis of small molecules with the ability to spontaneously self-organize into well-defined supramolecular architectures under a defined set of physical conditions. Over the past several years we have developed and utilized a new class of heteropolycyclic molecules to explore hydrogen bonding in water, self-replication in auto-catalytic systems, supramolecular chirality, and the underlying physical phenomena of self-assembly and self-organization processes. With this knowledge in hand, we were able to tailor the chemical, physical, and biological properties of 1-D tubular nanostructures for applications in the emerging fields of organic electronics, photovoltaics, nanobiotechnology, and nanomedicine. This lecture is an overview of the design, synthesis, and physical characterization of self-assembled organic nanotubes, and their biomedical applications.

Resume : Combining modern methods in microsystem technology with the latest advancements in the life sciences, namely those in tissue engineering and advanced cell culturing, is promoting the development of a promising toolbox for modeling biological systems. The core problem to solve using this toolbox is the design of 3D artificial cellular environments, both in fluidic systems and on solid substrates. The construction of 3D cell cultures on substrates involves various fabrication techniques which use different polymers and biopolymers processed by micromachining, chemical pattern guided cell cultivation, photopolymerization, and organ printing methods. These methods together have the potential to create an artificial system with the complete hierarchical, geometrical, and functional organization found in an actual biological system [1]. Starting from our work concerning 3D cultivation as a first step towards more biological modelling of cellular environments [2] we performed different studies concerning the use of such systems as pharmacological test platforms [3], the different genetic behaviour of the cells in 2 D and 3D [4], different structuring techniques [5, 6] towards the integration of different organ like systems on one chip [7]. In this contribution we will explain our approach to gain complex cellular structures while using chemical and mechanical modification of thin polymer foils. Due to folding and stacking of this pre manufactured cell sheet layers it is possible to achieve complex cellular and fluidic entities which are integrated in micro bioreactor systems.With the combined application of different methods it is possible to mimic complex tissue like structures of different organs. Preferable with the liver lobe we demonstrate the construction schema of such a multilayer techniques.We will discuss the different methods of designing such structures. Open questions of co-cultivation different cell types, structuring Disso space like areas and the methods thereof will be analyzed and conclusions are given. The liver plays a crucial role for the metabolism of both nutrients and drugs. Understanding and modelling of organomimetic cultivation substrates is a key technology with high potential for future developments in pharmaceutical drug discovery and tissue engineering.[1] A. Schober, U. Fernekorn, S. Singh, G. Schlingloff, M. Gebinoga, J. Hampl, A. Williamson, “Biotechnical multi-scale engineering mimicking the biological world ”, Eng. Life Sci. 2013, 13, 352–367;[2] A. Schober, C. Augspurger, G. Schlingloff, M. Gebinoga, M. Worgull, M. Schneider, C. Hildmann, U. Fernekorn, F. Weise, J. Hampl, L. Silveira, I. Cimalla, B. Lübbers, “Microfluidics and Biosensors as Tools for NanoBioSystems research with applications in the Life Science”, Materials Science and Engineering: B 169 (2010), 174-181;[3] U. Fernekorn, J. Hampl, F. Weise, C. Augspurger, C. Hildmann, M. Klett, A. Läffert, M. Gebinoga, K.F. Weibezahn, G. Schlingloff, M. Worgull, M. Schneider, A. Schober, Microbioreactor design for 3-D cell cultivation to create a pharmacological screening system; Eng. Life Sci. 2011, 11, No. 2, 133-139;[4] Fernekorn U., Hampl, J., Augsburger C., Hildmann Ch., Weise F., Klett M., Läffert A., Gebinoga M., Williamson A, Schober A., In vitro cultivation of biopsy derived primary hepatocytes leads to a more metabolic genotype in perfused 3D scaffolds than static 3D cell culture, RSC Adv., 2013, 3, 16558-16568;[5] M. Gebinoga, J. Katzmann, U. Fernekorn, J. Hampl, F.Weise, M. Klett, A. Läffert, T. Klar, A. Schober, “Multiphoton structuring of native polymers: A case study for structuring natural proteins”, Eng. Life Sci. 2013, 13,, Issue 4, pages 368–375;[6] J. Tobola, Schober et al. „Thermoforming techniques for manufacturing porous scaffolds for application in 3D cell cultivation” to be submitted;[7] Williamson A., Singh S., Fernekorn U., Schober A., The future of the patient-specific Body-on-a-chip, Lab Chip, 2013, 13, 3471-3480 The Royal Society of Chemistry 2013

Resume : emperature-responsive cell culture surfaces (TRCS), which is tissue-culture polystyrene (TCPS) modified with poly(N-isopropylacrylamide) (PIPAAm) (PIPAAm-TCPS), has been used for the fabrication of various types of single monolayer sheets such as epithelium keratinocyte, corneal epithelial cell, oral mucosal epithelial cell, urothelial cell, periodontal ligament, cardiomyocyte, hepatocyte cell sheets, and so on. Various types of cell sheets have been directly transplanted to reconstruct tissue, and some of them are clinically applied. PIPAAm-TCPS has been conventionally prepared by using electron beam (EB) irradiation and is currently released as commercially available products. However, the competitive price is desired for the commercially available PIPAAm-TCPS because of the expensive price. And also a lot of researchers would like to have an opportunity to prepare and investigate characteristic of TRCS by themselves. More facile methods for the preparation of PIPAAm-TCPS have been alternatively required without the expensive and special EB irradiation. In addition, highly-functional TRCS have been developed for the fabrication of elaborate cell sheets. In this presentation, we will show facile and alternative method for the preparation of TRCS and the recent development of highly-functional TRCS.

Resume : Nature has found cryptobiotic ways to preserve genetic information and protect cellular components against external stresses, such as nutrient deprivation, desiccation, high temperatures, radiation, and caustic chemicals. For example, a bacterial endospore, usually formed in response to nutrient deficiency, is a non-dividing, dormant body, which possesses a thin but tough proteinaceous coat at its outmost layer. Beneath the coat is a buffering cortex layer of peptidoglycan. This hierarchical shell structure allows the endospore to survive for many years (in some cases, up to millions of years) under hostile conditions found naturally that can easily and quickly kill normal cells. Recent studies have sought to chemically control and tailor the metabolic behaviors of non-spore-forming cells, as well as enhancing their viability against adverse environmental conditions, by forming thin (<100 nm), tough artificial shells. These living "cell-in-shell" structures, called artificial spores (chemically-formed spore-like structures), enable control of cell division, protection against physical and chemical stresses, and cell-surface functionalizability, as well as providing the cells with exogenous properties that are not innate to the cells but are introduced chemically, such as magnetism, heat-tolerance, and UV-resistance.

Resume : Molecularly imprinted polymers (MIPs) are synthetic antibody mimics ('plastic antibodies') that specifically recognize molecular targets.1-3 They are highly cross-linked polymers that are synthesized through the polymerization of monomers bearing suitable functional groups, in the presence of the target molecule acting as a molecular template. This templating induces three-dimensional binding sites in the polymer that are complementary to the template in terms of size, shape and chemical functionality. Thus, the plastic antibody can recognize and bind its target with an affinity and selectivity similar to a biological antibody. We present here a new approach allowing for the synthesis of MIP by controlled/living radical polymerization using a dendritic multiiniferter.4 This results in protein-size, soluble MIP nanogels with a homogeneous size distribution. Their mean diameter is 17 nm and the average molecular weight 97 kDa, thus their size and density are very close to those of biological antibodies. The MIP nanogels show specific binding of their targets, small organic molecules or proteins, with a nanomolar affinity and a good selectivity. We also present new methods to prepare MIPs specific for proteins based on solid-phase synthesis around the immobilised template,5 or in solution using anchoring monomers.6 In addition, the direct coating of thin MIP films around functional inorganic cores (magnetic nanoparticles,7 quantum dots, upconverting nanoparticles8) by localized photopolymerization will be described. The use of these functional nanomaterials for enzyme inhibition,6 enzyme stabilization, for sensing,9 and for cell and tissue imaging will be discussed. [1]. R, Arshady, K. Mosbach, Makromol. Chem. 1981, 182, 687-692. [2]. G. Wulff, A. Sarhan, Angew. Chem. Int. Ed. 1972, 11, 341. [3]. K. Haupt, A.V. Linares, M. Bompart, B. Tse Sum Bui, Top. Curr. Chem. 2012, 325, 1-28. [4]. P. Çakir, A. Cutivet, M. Resmini, B. Tse Sum Bui, K. Haupt, Adv. Mater. 2013, 25, 1048-1051. [5]. S. Ambrosini, S. Beyazit, K. Haupt, B. Tse Sum Bui, Chem. Commun. 2013, 49, 6746-6748. [6]. A. Cutivet, C. Schembri, J. Kovensky, K. Haupt, J. Am. Chem. Soc. 2009, 131, 14699-14702. [7]. C. Gonzato, M. Courty, P. Pasetto, K. Haupt, Adv. Funct. Mater. 2011, 21, 3947-3953. [8]. S. Beyazit, S. Ambrosini, N. Marchyk, E. Palo, V. Kale, T. Soukka, B. Tse Sum Bui, K. Haupt, Angew. Chem. Int. Ed. 2014 (in press, DOI: 10.1002/anie.201403576). [9] M. Bompart, Y. De Wilde, K. Haupt, Adv. Mater. 2010, 22, 2343-2348.

Resume : Synthetic octacalcium phosphate (OCP) displays a highly osteoconductive property which is induced by stimulating osteoblastic cells to differentiate through a progressive conversion process from OCP to hydroxyapatite (HA) [1]. However, OCP cannot be sintered under maintaining the original crystal phase due to the inclusion of large amount of water molecules in the structure. We have shown that the moldability and implantability can be markedly improved by combining OCP with several natural polymers. The addition of hyaluronic acids (HyAs) allows OCP granules, having several hundreds μm diameter, to form hydrogel-like materials and be injectable [2]. OCP/HyAs can be relatively easily injected into PTFE ring, a model space of bone augmentation, placed onto mouse calvaria using a syringe. The osteocondutivity of OCP/HyAs was significantly higher than that of OCP granules alone but their biological performances were dependent on the molecular weight of HyAs used. The mixing OCP with gelatin (Gel) molecules and the cross-linking of the Gel matrix allows OCP to be porous and 3D form in which the host osteoblastic cells in vivo could be easily invaded [3]. OCP can be co-precipitated with Gel molecules in a specified condition so that the OCP crystals were dispersed highly homogenously in the Gel matrix. The effect of OCP/Gel implantation was confirmed in rat calvaria critical-sized defect and in rabbit tibia defect, which showed a highly biodegradable property of OCP/Gel coupled with an active new bone formation [3,4]. Thus, OCP-based materials can easily be molded for the designed shape to substitute such bone defects and show highly osteoconductivity. The biological performance and the handling property of OCP-based materials we developed so far will be presented along with the elucidated mechanisms to show the osteoconductivity from the view point of biomaterial science. References: [1] Suzuki O et al. Biomaterials 2006, 27:2671, [2] Suzuki K et al. Acta Biomater 2014, 10:531, [3] Handa T et al. Acta Biomater 2012, 8:1190, [4] Suzuki K et al. Phosphorus Res Bull, 2012, 26 (Special Issue) 53.

Resume : Calcium Phosphate Cements (CPCs) are more and more used as bone substitutes, but a number of technological issues still need to be addressed, in particular on the improvement of mechanical properties. The present work aims at reviewing recent works on the links between microstructure (porosity at different scales, nature and amount of polymer additives…) and several mechanical properties, may they concern elastic behaviour (Young’s modulus) or fracture behaviour (toughness and tolerance to damage), in relation with processing (strength and its statistical Weibull analysis). Ways to produce a variety of microstructures will be described (changing the liquid-to-powder ratio in the cement paste, incorporating porogenic agents, adding cellulose ethers and derivatives…) and their influence on both handling properties (injectability, cohesion…) and mechanical properties will be discussed, in an attempt to propose strategies for material optimisation. In particular, it is shown that the addition of some polymeric additives can improve fracture toughness, induce a good tolerance to damage, give the cement paste excellent injectability and cohesion at early age, and lead to interesting porous microstructures in view of biomedical applications.

Resume : Biomaterials nanocharacterization fundamentals are based on surface electronic structure testing and nanoimaging atoms at a surface and morphology due to adaptation of ultrahigh vacuum scanning probe microscope JSPM-4610,JEOL and NMR and EPR,BRUKER spectrometers In result ,has been developed methodics of a research and practical applications in innovative nanotechnology, for example, biomimetic nanoparticles of bioceramics.. In the SPM&RS-Centre, there is the capability to use for nanocharacterization such methods as scanning tunneling microscopy, tunneling and magnetic spectroscopy, atomic force microscopy, X-ray photoelectron spectroscopy, nuclear magnetic resonance and electronic paramagnetic resonance, which allow to study at qualitatively new level such physical quantities as the DOS distribution, electron work function distribution, volt-ampere characteristics, surface conductance, atomic and electronic structure, electron-nuclear system dynamics, and to visualize individual atoms on the surface and morphology of a surface and nanoparticles for nanobiomaterials.

Resume : Cell adhesion-associated proteins, which include integrin family, focal adhesion kinase, paxillin, vinculin, zyxin, talin, Src, actin, and etc., are the important components in controlling the cell behavior, such as the intracellular and extracellular signal transduction, cell adhesion, motility and migration, embryonic development, tissue function, inflammation, and wound healing. Because these cell behaviors can be regulated by multiple adhesion-associated proteins, it is difficult to obtain the contribution from individual proteins using conventional optical imaging. In this report, we have investigated the formation of the cell adhesion on the nano-patterned extracellular matrix using Chinese hamster ovary (CHO) cells. When the cells attach or migrate on the extracellular matrix, such as the fibronectin and collagen, nano-patterned surfaces, the formation of focal adhesion can be manipulated by the extracellular matrix nano-patterns. However, the conventional optical imaging cannot be used to visualize the individual proteins on the fibronectin nano-patterned surface, due to the optical diffraction limit. Therefore, the multi-color super-resolution localization imaging was used to investigate the response of the cell adhesion-associated proteins on the fibronectin nano-patterned surfaces. The spatial correlation of the cell adhesion-associated proteins has been investigated by super-resolution fluorescence imaging with a few tens of nanometers spatial resolution. Combining the protein pair correlation analysis, it quantitatively provides the molecular information of the cell adhesion-associated proteins in the different types of focal adhesions. We will also present the studies of aritifical ceullar environements by advanced imaging tools such as AFM, Raman microscpy, nonlinear optical imaging, and TOF-SIMS.

Resume : To the purpose of modeling processes after implantation of bioactive synthesized HAp ceramic and to evaluate bioadhesion to cell culture, biofilm growth and cytotoxicity of domestic bioactive ceramic on different stages of their interaction with the environment of tissue in vitro studies have been carried out using cell cultures. The conventional method of culturing cells have been used with HEP- 2 cells in DMEM culture medium (Sigma, USA) and with addition of penicillin, streptomycin and 5% fetal calf serum in 24 well plates in an incubator at 37 ° C in atmosphere with 5% СО2.. The replacement of the culture medium performed daily. Powder of composed nanoparticles of nanocomposite hydroxyapatite (HAp) - tricalcium phosphate (TCP) (80 : 20%) (particle has size 25 - 40 nm, granules - 0.3 -0.5 mm, pore - 50 - 150mkm) has been synthesized. In two parallel experiments the freshly calcite ceramics have been used or the same ceramics that was modified by overnight exposure in helofuzyn. The cells investigation by the microscopy and fluorescent inverted microscope methods carried out. All experiments showed tight adhesion of cells to ceramic and active multilayered cell growth at the surface and near area. It was reveal what the fresh calcite ceramics inhibit of cell growth was observed during the first 1.5 - 2 days compared to the ceramics modified by helofuzyn. Fluorescent study also found a higher level of luminescence of nuclei and cytoplasm of cells near and on the modified ceramics at the first few days of cultivation, reflecting the increase in their proliferate activity. These features can be explained by the active adsorption at the surface of fresh calcite ceramics of components of the culture solution and the excretion of some elements from these surfaces accumulated on them due to segregation processes during calcination. These processes are especially significant during first time after implantation and are accompanying by local changes in pH and ionic composition of the solution, which causes an initial delay in cell growth.

Resume : We have previously shown that synthetic octacalcium phosphate (OCP) combined with hyaluronic acid (HyA) has an injectable characteristic and enhances osteoconductive property more than that by OCP alone [1]. In the present study, we investigated the effect of HyA and OCP on the osteoclastic differentiation of murine macrophage RAW264 cells in vitro. 48-well plastic culture plate was prepared by coating the particles of OCP. Sodium hyaluronic acid medical products with different molecular weights, 9x105 (HA 90), 19x105 (HA 190) and 60x105 (a chemically-modified derivative of sodium HyA, HA 600) were used as the additives in the cell culture medium. Under the stimulation of RAW264 cells with RANKL, the appearance of TRAP positive multinuclear cells increased in the presence of HyAs when cultured without OCP coating as reported previously [1]. The significant increase was observed in the presence of HyA 90 or HyA 600 [1]. When the cells were cultured on OCP coating, although the TRAP staining was apparent as reported in the co-culturing of osteoblasts and osteoclast precursor cells [2], the staining seemed to be enhanced by the HyAs treatment in the same manner without OCP. It is probable that the enhancement of these HyAs on the osteoclast differentiation in vitro is associated with increasing the osteoconductive property of OCP with HyAs observed in the previous in vivo study [1]. These results suggest that HyAs may have an influence on the osteoconductivity of OCP by up-regulating the osteoclastic resorption activity. References: [1] Suzuki K et al. Acta Biomater 10:531-543, 2014; [2] Takami M et al. Tissue Eng Part A 15:3991-4000, 2009.

Resume : Contemporary technology enables to create a synthetic bone graft that rivals their capabilities of natural origin biomaterials. However, it is possible to modify the synthetic material to increase biocompatibility and regulate biodegradation time and add new osteostimulation, angiogenesis, anti-inflammatory properties, etc. We have been planned and synthesized biphasic calcium phosphate ceramics with crystals range in several tens nanometers. The biphasic ceramics (30%) was combined with glass-ceramics (70%) selected composition to form the bioactive ceramic composite (BCC). During a synthesis, the bioceramic composite granules (0,3-0,8 mm) were directly modified by additional doping with silver and copper ion's combination at 2:1 ratio within 10 at% and 5 at% range accordingly. The aim was to explore the antimicrobial activity and to investigate the cytotoxic properties of the modified bioceramics in vitro. The cytotoxicity tests have been conducted in human adipose-derived mesenchymal stromal cell (MCSs) culture to investigate the biocompatibility of the BCC samples. The antimicrobial properties have been determined by the degree of stunting bacterial cultures. As a test-microorganisms have been used the strains of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. It was determined extracts of the modified ceramic samples had no significant effect on cell proliferation within 1 at% Ag+ and 0,5 at% Cu2+ range. Meanwhile, the BCC sample's granules in such ion concentration in direct cytotoxicity test caused cell death in culture. By vital fluorescein diacetate staining and fluorescence microscopy it was confirmed the adhesion of MSCs to the modified bioceramic's granules. It was found that the synthetic implant material doped with silver and copper ions starting from 0,5 at% Ag+ and 0,25 at% Cu2+ range revealed the antibacterial and antimycotic effects, which depend on additions concentration. Selected bioceramic composite doped with a combination of silver and copper ions within 1 at% and 0,5 at% range respectively does not exhibit cytotoxic effect and does not inhibit MSCs adhesion and provides moderately expressed antimicrobial activity.

Resume : Biochips are attracting large interest in cell biology as functional tools to perform quick and extensive cell studies by integrating different functions in a single chip. In this respect, impedance spectroscopy (IS) is an emerging read-out technique since the immobilization/adhesion of cells on biofunctionalized ceramic scaffold to the cell before bone tissue regeneration. The resistance are correlated to cell number, adhesion and cytoskeleton organization. These biochips are very cheap and reusable and represent a robust method to count cells with great sensitivity without detaching/destroying them (a crucial property for further assays such as migration tests and/or cytotoxicity tests): In addition we have used the biochip to monitor the adhesion between two different cell type, endothelial cells and leukaemia cells.

Resume : In developing optical biosensors, whose work is based on the determination of the spectral absorption characteristics of organic matter, it is necessary to apply cutting optical filters for the selection of certain infrared ranges of the spectrum. The work carried out to develop multilayer thin film interference filters with replaceable cutting position limits infrared radiation depending on the concentration and type of the studied compounds. The peculiarity of these filters is using semiconductor crystals In4Se3 and In4Te3 as substrates for thin-film multilayer coating. Crystals In4Se3 and In4Te3 are layered and well separated by cleavage planes (100), which allows us to produce plates required for the optical cell thickness without cutting and processing operations. The fundamental absorption, which is caused by the corresponding band gap (0.65 eV for In4Se3 to 0.48 eV for In4Te3), provides the absorption component of the functioning of the filter. High spectral characteristics, stability, resistance to chemicals and atmospheric single crystals In4Se3, In4Te3 enable design based on these filters with = 1,7-6,5 um different positions of shortwave radiation limits cutting range and a maximum transmittance Tmax = 91-95%. The method of SEM studies depending on the structure of thin films - component filters ZnS, SiO, Ge, Te, SrF2 on technological factors and the effect of organic compounds on spectral and mechanical stability of the multilayer coating. The design of the active cell optical biosensor developed using IR filters.

Resume : In last decades biorelated nanocomposites become driving force for huge number of investigations. For example combination of the magnetic nanowires with bioactive molecules leads to an interesting hybrid system which combine properties of nanostructures and bioparticle in one spices. In such manner specific recognition or catalytic properties of biomaterials are convoluted with the attractive electronic, optical, magnetic and structural characteristics of magnetic nanowires. However, to obtain biocomposite, nanostructures should be properly functionalized what can be realized by various linkage chemistries. Functional compounds can be directly bonded with organic molecules or via connectors like glutaraldehyde. The other option is noncovalent interactions with fatty acids, proteins.. Different ligands with different affinity both to the modified surfaces and bioparticles can be obtained: -SH, -COOH, -OP, -CN. The reason why magnetic nanostructures/nanowires are considered as a promising candidate of biocomposite constituents is their magnetic properties, which gives fast and easy manipulation tool with use of external magnetic field. Structural and magnetic properties of magnetic nanoparticles and composites will be tested by: XRD, TEM, IR and Mössbauer spectroscopy.

Resume : Protein-based scaffolds are actively investigated for the controlled assembly of advanced multifunctional materials taking advantage of the best of both inorganic and biological building blocks [1]. Due to their hybrid nature and novel properties, such materials have the potential to answer the current challenges and needs in applications including biosensing or regenerative medicine. In this study, amyloid protein nanofibres prepared from hen egg white lysozyme are used to specifically mediate the assembly of gold nanoparticles into regular arrays from solutions to different silicon oxide surfaces. The effect of nanoparticle size and deposition conditions on the self-assembly process is investigated by means of atomic force microscopy (AFM) and UV-visible spectroscopy (UV-Vis). The periodic nanoparticle arrangement observed is mainly influenced by inter-particle interactions rather than by the presence of specific binding groups at the surface of the template. For a fixed ionic strength and particle concentration, the distance between nanoparticles increases with nanoparticle diameter and this trend fits well with calculated interparticle potential values. In turn, optical properties measured by UV-Vis on large surfaces are correlated with particle arrangements observed at the nanoscale. 1. F. Leroux, M. Gysemans, S. Bals, K. J. Batenburg, J. Snauwaert, T. Verbiest, C. Van Haesendonck, G. Van Tendeloo, Adv. Mater., 2010, 22(19), 2193-7.

Resume : Laser-induced periodic surface structures (LIPSS) are frequently observed coherent structures in laser surface processing. For polyethylene terephthalate (PET), LIPSS can be induced by irradiation with linearly polarized ns-pulsed UV laser light at fluences well below the ablation threshold. The lateral period of the observed LIPSS is close to the wavelength of the laser light divided by the effective index of refraction neff (about 1.3). Gold or silver nanowires can be formed on top of the polymer structures by means of evaporation under an inclined angle. These separated nanowires exhibit localized surface plasmon resonances (SPR) in the VIS to NIR spectral region, overlapping with the so-called biological window. This together with the alignment of human cells on such LIPSS structures renders our nanowires as very promising candidates for biosensing applications.

Resume : Transition metal oxides like WO3 and MoO3 are versatile materials investigated nowadays for their properties like gas sensing and electrochromism. There are indications that both MoO3 and WO3 also possess antimicrobial properties, which can be used against proliferation of nosocomial infections in hospitals [1]. In this work two new approaches are presented: the preparation and characterization of Mo-W oxides combinatorial thin film libraries and the proof of principle as they can be used as antimicrobial coatings with screening compositional ranges active against E. Coli. Combinatorial thin film coatings with compositional spreads ranging from 91 to 44 at.% Mo (9 to 56 at.% W, respectively) were successfully deposited by thermal co-evaporation. After a multiple step heat treatment, a mixture of crystalline molybdenum and tungsten oxides and sub-oxides, and amorphous tungsten oxide were obtained. Preliminary tests of the coatings antimicrobial activity against E. Coli strain BL21DE3 (kanamycin resistant) were performed, by investigating the bacterial growth both onto the surface and in the volume of the working culture. In some certain compositional ranges the antimicrobial activity was proven, and some possible mechanisms were proposed. The mechanism involves a combined contribution of surface structuring and the presence of W19O49 phase [2]. [1] C. Zollfrank et al., Mater Sci Eng C 32, 47 (2012) [2] C. C. Mardare and A. W. Hassel, ACS Comb Sci – under review

Resume : Recently bioinspiered nanocomposites become driving force for many interesting investigations. Among others, integration of the magnetic nanoparticles with biomolecules leads to a novel hybrid system which connect characteristic of nano- and bioparticle in one. Such way can be combined recognition or catalytic properties of biomaterials with the attractive electronic, optical, magnetic and structural characteristics of magnetic nanoparticles. To obtain biocomposite, nanoparticles should be functionalized with adequate biomolecules through different linkage chemistries. It can be directly bonded with organic molecules which form at the surface self – assembled monolayers. These can be further modified either: by formation of covalent bonds with connectors like glutaraldehyde or by direct noncovalent interactions with fatty acids, proteins. Noncovalent bonds are one of the most important bonds in biological systems especially in hydrogen bonding networks. Different ligands with different affinity to the modified surfaces and bioparticles can be obtained: -SH, -COOH, -OP, -CN. The reason why magnetic nanoparticles are considered as a good candidate of biocomposite constituents is their magnetic properties, which gives fast and easy manipulation tool with use of external magnetic field. Structural and magnetic properties of magnetic nanoparticles and composites will be tested by: XRD, TEM, IR and Mössbauer spectroscopy.

Resume : One of the main challenges in nanomedicine is to provide early diagnosis, minimally-invasive and efficient treatments against cancer.In this context, iron oxide nanoparticles(IONP) are a real breakthrough in biomedical research.We review the current research on IONP to achieve “therasnostic” tools, with customized biological, chemical and physical functionalities for selective targeting detection and elimination of cancer cells1.A review of the fundamentals and experimental data will be shown, paying special attention to the influence of biological matrix on the nanomagnetism of IONP. This is crucial to warrant the heat mediated by IONP and its contrast agent signal for acting as efficient theranostic agents. Different results will be shown related to the IONP chemical synthesis, characterization of IONP dynamic magnetic and thermal properties2 for generating efficient intracellular hyperthermia3, their bio-functionalization with anticancer agents, and biological studies related to the IONP internalization and anticancer activity4.New guidelines to optimize the heating efficiency of IONP via tailoring heating mechanisms will be shown5,opening the way towards improving the use of IONP in biomedical applications. [1]F.J.Teran,et al “Cancer treatment using magnetic nanoparticles”.Yearbook of Science & Technology 2013, McGrawHill. [2] J. Mater. Chem. 22,21065 (2012); [3] Appl.Phys.Lett.101,062413 (2012); [4]Nanomedicine 10,733 (2013), [5] Salas et al. submitted to J. Phys.Chem.C

Resume : An optically transparent poly(3,4-ethylenedioxythiophene) (PEDOT) based organic electronic devices have been developed to investigate the behavior of human mesenchymal stem cell (hMSC) and to sense and to capture circulating tumor cells. We first conducted a control experiment on electrical cell-substrate impedance sensing (ECIS) device by culturing the hMSC on the chip. According to our result, the impedance increased reflected the hMSC proliferation, attachment and motility during the first 16 hours of cell culture. In order to control the differentiation of human mesenchymal stem cell (hMSC) on chip, we also developed all-solution-processed multifunctional organic devices, comprising reduced graphene oxide (rGO) and dexamethasone 21-phosphate disodium salt (DEX) drug loaded poly(3,4-ethylenedioxythiophene) (PEDOT) microelectrode arrays on indium tin oxide glass, that can be used to manipulate differentiation. In our devices, the rGO micropatterns were used as the adhesive coating to attract the adhesion of hMSC cells whereas PLL-g-PEG coated PEDOT electrodes served as the anti-adhesive coating where no hMSC cells can attach. In addition, the PEDOT electrodes also work as drug releasing components where control DEX release from PEDOT matrix can be achieved via cyclic potential stimulation (CPS). To capture the circulating tumor cells, we fabricated 3D PEDOT-based micro/nanorod array, which can be further surface-grafted with capture agents for directed specific recognition to study the cell–substrate interactions on bioelectronics interfaces (BEIs). This BEI platform features the advantageous characteristics: (1) diverse dimensional structures (tunable from the microscale to the nanoscale), (2) varied surface chemical properties (tunable from nonspecific to specific), (3) high electrical conductivity, and (4) reversible chemical redox switching. Furthermore, through systematic studies of PEDOT systems, we explore the effects of both chemistry and topography on the circulating tumor cell (CTC)-capture performance. The 400 nm PEDOT pillars exhibited the optimal cell-capture efficiency; it could be used to isolate CTCs with minimal contamination from surrounding nontargeted cells (e.g., EpCAM-negative cells, white blood cells) and negligible disruption of the CTCs’ viability and functions. It is conceivable that PEDOT-based micro/nanorod array films function as a critical therapeutic intervention for monitoring tumor progression and metathesis, providing valuable insight into the use of electronics for tissue engineering and regenerative medicine.

Resume : Many natural chromophores have recently emerged as suitable semiconducting materials for (opto)electronic applications. The motivation for exploring such molecules is the realization of biodegradable and biocompatible electronics fabricated from cheap and nontoxic materials. We find that natural pigment-forming dye molecules, such as those from the indigo family, form highly-ordered thin films with excellent -stacking. Using such films, we have demonstrated field-effect transistors (FETs) and complementary-like circuit elements utilizing exclusively natural materials operating at the state-of-the-art level with respect to mobility and operational stability in ambient conditions. These dyes show air-stable ambipolar charge transport with balanced hole and electron mobilities in the range of 1×10-2 – 2 cm2/Vs. A very important property is that these molecules form high crystal lattice energy solids with exceptional operational, thermal, and chemical stability. FETs with indigos can be operated without any passivation in highly-demanding aqueous environments, within a pH range from 3-11 and with a variety of different ions. We report on the results of stressing tests in such underwater environments. The –NH and =O functional groups lend themselves to easy functionalizing, which we have exploited to two ends: 1) creation of stable colloidal nanocrystals with controllable morphology and optical properties, and 2) biofunctionalization. We discuss also preliminary experiments concerning field-effect transistor biodetectors based on these materials. Hydrogen-bonded natural and nature-inspired materials are an interesting and previously unexplored class of organic semiconductors with inherent potential for biointegrated applications.

Resume : Electrically conducting polymers (ECPs) were widely used to electrically couple with cells in implanted electronic devices and biosensors, mainly thanks to the long-term stability of their electrical properties in biological environments and their mechanical properties comparable with that of the extracellular matrix. To erase/diminish nonspecific interaction for ECPs would not only increase the selectivity of biosensors but also reduce the inflammation risk for implants. In this presentation, we show that with finely tuning the chemical structure, ECPs could specifically interact with cells on a protein-resistance background. Our recent works further demonstrated that with combining specific interaction with protein resistance on ECPs, they could differentiate cells, dynamically interact with and release cells, or even help to construct a three-dimensional platform to capture CTC cells with both high efficiency and selectivity.

Resume : The physical color in biological materials in certain cases is determined by the structural and optical properties of the constitutive photonic crystal. The most investigated examples are among arthropods where the colorful structures are located in scales [1]. Altering the refractive index of the materials will produce a measurable color change. As an application, butterfly wings in volatile vapors show concentration dependent optical reflectivity variation [2]. The refractive index change also depends on temperature and is enhanced by capillary condensation [3]. Here we will compare the behavior of butterfly wings as gas sensors with bioinspired samples. On biological detectors we have shown that different butterfly wings exhibit specific response to the same vapors [4]. Chemical selectivity has been shown for different vapors on the same nanostructure [2]. Now we intend to investigate nanoarchitectures with similar structural properties made of chitinous material and we will compare them to inorganic Si based nanocomposites. Recently we pointed out the effect of minor structural differences on the reflectance spectra of Blue butterfly wings [5], the same delicate dependence is expected in case of bioinspired structures. [1] LP Biró & JP Vigneron, Laser & Photonics Rev. 5, 27, 2011 [2] RA Potyrailo et al. Nat. Phot. 1, 123, 2007 [3] K Kertesz et al. Appl. Surf. Sci. 281, 49, 2013 [4] LP Biró et al. Proc SPIE 7057, 705706-1, 2008 [5] Zs Bálint et al. Interface 9, 1745-1756

Resume : Magnetoelectronics at the nanoscale provides new strategies to tackle issues concerning to life sciences and healthcare: from recognition of molecules to therapeutic health treatments. The basic requirement needed by this novel research field, is the strict synergy of a wide spectrum of scientific competences: physicists, chemists, biologists, physicians and engineers are called to join their expertise in order to realize innovative tools and new-concept devices. Magnetic structures can be exploited to detect molecules, bacteria and cells or to improve the functionalities of artificial biological tissues and finally may be directly employed to drug delivery and to kill cancers. However, this is only possible through a smart bio-functionalization of the magnetic objects, and a control onto their magnetic properties. I illustrate different examples of bio-sensing and bio-medical applications based on magnetic nanomaterials.

Resume : Natural animal silks from spider and silkworm have many remarkable properties. The dimension and functions of spinning apparatus in vivo were biomimicked in microfluidic devices with microchannels. Raman spectroscopy and small angle X-ray scattering (SAXS) technique were adopted to analyze the conformation transition of regenerated silk fibroin (RSF) and the shape parameters of RSF aggregates in aqueous solutions before and after flowing in the microchannel. Results showed that higher RSF concentration, lower pH value and higher concentration of calcium ion in the solution promoted the growth of RSF aggregates and the conformation transition of RSF from α-helix to β-sheet. The increasing shear time and elongation rate had similar effects on the structure transition of RSF in aqueous solution. We also integrated multiple functions of the natural spinning apparatus in the microfluidic device. These functions include protein composition adjustment, shearing and elongation of spinning dope and spinning. The integration may have significant influence on the application of microfluidic technology in bio-inspired spinning.

Resume : Circulating tumor cells (CTCs) have become an emerging “biomarker” for monitoring cancer metastasis and prognosis. Although there are existing technologies available for isolating/counting CTCs, the most common of which using immunomagnetic beads, they are limited by their low capture efficiencies and low specificities. By introducing a three-dimensional (3D) nanostructured substrate – specifically, a silicon-nanowire (SiNW) array coated with anti-EpCAM – we can capture CTCs with much higher efficiency and specificity. The conventional methods of isolating CTCs depend on biomolecular recognitions, such as antigen-antibody interaction. Unlikely, we here proposed that nanoscaled local topographic interactions besides biomolecular recognitions inspired by natural immuno-recognizing system. This cooperative effect of physical and chemical issues between CTCs and substrate leads to increased binding of CTCs, which significantly enhance capture efficiency. Recently, we have also developed a 3D cell capture/release system triggered by aptamer enzyme, electrical potential and Temperature, which is effective and of “free damage" to capture and release cancer cells. The bio-inspired interfaces of cell capture and release open up a light to rare-cell based diagnostics, such as CTCs, fetal cells, stem cell and so on.

Resume : Aptamers are nucleic acid sequences that could selectively bind to their target molecules similiar to antibodies [1,2]. The synthesis and selection of aptamers are done by using SELEX (Systematic Evolution of Ligands by Exponential enrichment) method. The improved stability properties including resistant to denaturation and degradation, easy modification, target adaptability and easy-to-stock make them more advantageous comparison to antibodies and ideal candidates as protein recognition elements in a wide range of bioassays and for diagnostic applications. The development of advanced biosensor platforms could impact significantly the areas of genomics, proteomics, biomedical diagnostics and drug discovery [3-5]. Electrochemical biosensors coupling the inherent specifity of biorecognition reactions with the high sensitivity of physical transducers, hold great promise for detection of sequence-specific nucleic acids, or proteins for clinical, environmental or forensic investigations [2-5]. An overview to electrochemical monitoring of aptamer-protein interactions in combination with disposable sensor platforms has been presented herein with their advantages and further applications. Acknowledgements. A.E acknowledges the financial support from Turkish Scientific and Technological Council (TUBITAK) (Project no.111T073), and she also would like to express her gratitude to the Turkish Academy of Sciences (TUBA) as the associate member of TUBA for its partial support. References 1- J. Muller, B. Isermann, C. Ducker, M. Salehi, M. Meyer, M. Friedrich, T. Madhusudhan, J. Oldenburg, G. Mayer, B. Potzsch, Chem. Biol. 16 (2009) 442. 2- M. Mascini, I. Palchetti, S. Tombelli, Angew. Chem. Int. Ed. 51 (2012) 1316. 3- E. Palecek, M. Bartosík, Chem. Rev. 112 (2012) 3427. 4- J. Wang, Electroanalysis 17 (2005) 7. 5- A. Erdem, Talanta, 74 (2007) 318.

Resume : Prospects for practical application of semiconductor sensors based on nanostructured and biointegrated materials primarily related to the development of so-called advanced cyber-physical systems. These systems are based on the use of intelligent embedded computer devices and are designed to analyze large volumes of incoming information signals in real-time a set and synthesis of recommendations to support decision-making, or forming the output array control signals to ensure the stability of the system under uncertainty. The variety of the analyzed signal generated by biomedical, biotech, telemetry and other systems led to the need to formulate a new direction - multifunctional embedded selfreconfigurable systems able to adapt to the solvable problem-oriented tasks. Clearly, reliability and portability of such systems is determined by the correctness and reliability of the signal processing of semiconductor, including optical, electrical, biointegrated and others sensors, which are the basis of information-measuring systems tract of the cyber-physical systems. In this research the technique and conducted testing of the synthesis of multifunctional selfreconfigurable embedded computer systems based on the through systematic analysis of a set of problem-oriented tasks for which the synthesized system. The basis of this analysis is the choice of the set of measured parameters characteristic of physical objects or processes with appropriate descriptions of arrays, such as temperature, current, voltage, light power and more: M = {[T, U, I, F, ...] [vT, VD, ...]}. Approved structural and algorithmic solutions for the implementation of selfreconfigurable directly depend on the boundary, say, the control parameter values of the object or process error computations that are crucial in selecting methods and algorithms for synthesizing information and identifies the parameters used sensors - current, photo- concentration sensitivity, temperature range, and others. As a simulation model for the implementation of the proposed principles of synthesis of selfreconfigurable embedded multifunctional computer system tested information-measuring system implemented on top of the pH sensor and gas sensor CO, SxOy, NxOy.