Primary human lung pericytes support and stabilize in-vitro perfusable microvessels.

The formation of blood vessels is a complex tissue-specific process that plays a pivotal role during developmental processes, in wound healing, cancer progression, fibrosis and other pathologies. To study vasculogenesis and vascular remodeling in the context of the lung, we developed an in-vitro microvascular model that closely mimics the human lung microvasculature in terms of 3D architecture, accessibility, functionality and cell types. Human pericytes from the distal airway were isolated and characterized using flow cytometry. To assess their role in the generation of normal microvessels, lung pericytes were mixed in fibrin gel and seeded into well-defined microcompartments together with primary endothelial cells (HUVEC). Patent microvessels covering an area of 3.1 mm2 formed within 3-5 days and were stable for up to 14 days. Soluble signals from the lung pericytes were necessary to establish perfusability, and pericytes migrated towards endothelial microvessels. Cell-cell communication in the form of adherens and tight junctions, as well as secretion of basement membrane was confirmed using transmission electron microscopy and immunocytochemistry on chip. Direct co-culture of pericytes with endothelial cells decreased the microvascular permeability by one order of magnitude from 17.8∙10-6 cm/s to 2.0∙10-6 cm/s and led to vessels with significantly smaller and less variable diameter. Upon phenylephrine administration, vasoconstriction was observed in microvessels lined with pericytes but not in endothelial microvessels only. Perfusable microvessels were also generated with human lung microvascular endothelial cells and lung pericytes. Human lung pericytes were thus shown to have a prominent influence on microvascular morphology, permeability, vasoconstriction and long-term stability in an in-vitro microvascular system. This biomimetic platform opens new possibilities to test functions and interactions of patient-derived cells in a physiologically relevant microvascular setting.

Self-assembly into nanoparticles is essential for receptor mediated uptake of therapeutic antisense oligonucleotides

Antisense oligonucleotides (ASOs) have the potential of revolutionizing medicine due to their ability to manipulate gene function for therapeutic purposes. ASOs are chemically modified and/or incorporated with nanoparticles to enhance their stability and cellular uptake; however, one of the biggest challenges is the poor understanding of their uptake mechanism, which is needed for designing better ASOs with high activity and low toxicity. Here, we study the uptake mechanism of three therapeutically relevant ASOs (peptide-conjugated phosphorodiamidate morpholino (P-PMO), 2?Omethyl phosphorothioate (2?OMe) and phosphorothioated tricyclo DNA (tcDNA) that have been optimized to induce exon skipping in models of Deuchenne muscular dystrophy (DMD). We show that P-PMO and tcDNA have high propensity to spontaneously self-assemble into nanoparticles. P-PMO forms micelles of defined size and their net charge (zeta potential) is dependent on the medium and concentration. In biomimetic conditions and at low concentrations P-PMO obtains net negative charge and its uptake is mediated by class A scavenger receptor subtypes (SCARAs) as shown by competitive inhibition and RNAi silencing experiments in-vitro. In-vivo, the activity of P-PMO was significantly decreased in SCARA1 knock-out mice compared to wild-type animals. Additionally, we show that SCARA1 is involved in the uptake of tcDNA and 2?OMe as shown by competitive inhibition and co-localization experiments. Surface plasmon resonance binding analysis to SCARA1 demonstrated that P-PMO and tcDNA have higher binding profiles to the receptor compared to 2?OMe. These results demonstrate receptor-mediated uptake for a range of ASO chemistries, a mechanism that is dependent on their self-assembly into nanoparticles.

Repair of Annulus fibrosus with Genipin-enhanced Fibrin Hydrogel and silk membrane- fleece

Introduction Low back pain is often caused by a trauma causing disc herniation and /or disc degeneration. Although there are some promising approaches for nucleus pulposus repair, the inner tissue of the intervertebral disc (IVD) so far no treatment or repair is available for annulus fibrosus (AF) injuries. Here we aimed to develop a new method to seal and repair AF injuries by using a silk fleece composite and a genipin enhanced hydrogel. Methods Bovine (b) IVDs were harvested under aseptic conditions and kept in free swelling conditions for 24h in high-glucose DMEM containing 5% bovine serum for equilibration (1). A circular 2mm biopsy punch (Polymed Medical Center, Switzerland) was used to form a reproducible defect in the AF. For filling the defect and keeping the silk composite in place a human-derived fibrin gel (Baxter Tisseel, Switzerland) enhanced with 4.2mg/ml of the cross linker genipin (Wako Chemicals GmbH, Germany) was used. The silk composite consists of a mesh- and a membrane side (Spintec Engineering GmbH, Germany); the membrane is facing outwards to form a seal. bIVDs were cultured in vitro for 14 days either under dynamic load in a custom-built bioreactor under physiological conditions (0.2MPa load and ±2° torsion at 0.2Hz for 8h/day) or static diurnal load of 0.2MPa (2). At the end of culture discs were checked for seal failure, disc height, metabolic activity, cell death by necrosis (LDH assay), DNA content and glycosaminoglycan content. Results Silk composite maintained its position throughout the 14 days of culture under loaded conditions. Although repaired discs performed slightly lower in cell activity, DNA and GAG content were in the range of the control. Also LDH resulted in similar values compared to control discs (Fig 1). Height loss in repaired discs was in the same range as for static diurnal loaded control samples. For dynamically loaded samples the decrease was comparable to the injured, unrepaired discs. Fig 1 LDH of repaired discs compared to control disc after 24h in free swelling conditions for equilibration and first three loading cycles. Conclusions Silk-genipin-fibrin reinforced hydrogel is a promising approach to close AF defects as tested by two degree of freedom loading. In further experiments cytocompatibility of genipin has to be investigated. References 1. Chan SC, Gantenbein-Ritter B. Preparation of intact bovine tail intervertebral discs for organ culture. J Vis Exp 2012, Feb 2;60(60):e3490. 2. Walser J, Ferguson SJ, Gantenbein-Ritter B. Design of a mechanical loading device to culture intact bovine caudal motional segments of the spine under twisting motion. In: Davies J, editors. Replacing animal models: a practical guide to creating and using biomimetic alternatives. Chichester, UK: John Wiley & Sons, Ltd.; 2012. p. 89-105. Acknowledgements This project is funded by the Gerbert Rüf Stiftung project # GRS-028/13 and the Swiss National Science Project SNF #310030_153411.

Dual Role of Mesenchymal Stem Cells Allows for Microvascularized Bone Tissue-Like Environments in PEG Hydrogels.

In vitro engineered tissues which recapitulate functional and morphological properties of bone marrow and bone tissue will be desirable to study bone regeneration under fully controlled conditions. Among the key players in the initial phase of bone regeneration are mesenchymal stem cells (MSCs) and endothelial cells (ECs) that are in close contact in many tissues. Additionally, the generation of tissue constructs for in vivo transplantations has included the use of ECs since insufficient vascularization is one of the bottlenecks in (bone) tissue engineering. Here, 3D cocultures of human bone marrow derived MSCs (hBM-MSCs) and human umbilical vein endothelial cells (HUVECs) in synthetic biomimetic poly(ethylene glycol) (PEG)-based matrices are directed toward vascularized bone mimicking tissue constructs. In this environment, bone morphogenetic protein-2 (BMP-2) or fibroblast growth factor-2 (FGF-2) promotes the formation of vascular networks. However, while osteogenic differentiation is achieved with BMP-2, the treatment with FGF-2 suppressed osteogenic differentiation. Thus, this study shows that cocultures of hBM-MSCs and HUVECs in biological inert PEG matrices can be directed toward bone and bone marrow-like 3D tissue constructs.

Study of Basic Wood Decay Mechanisms and Their Biotechnological Applications

The overall objective of this thesis was to gain further understanding of the non-enzymatic mechanisms involved in brown-rot wood decay, especially the role of pH, oxalic acid, and low molecular catecholate compounds on the dissolution and reduction of iron, and the formation of reactive oxygen species. Another focus of this study will be the potential application of a biomimetic free radical generating system inspired from fungi wood decay process, especially the non-enzymatic mechanism. The possible pathways of iron uptake and iron redox cycling in non-enzymatic brown-rot decay were investigated in this study. UV-Vis spectroscopy and HPLC were employed to study the kinetics and pathways of the interaction between iron and model catecholate compounds under different pH and chelator/iron molar ratio conditions. Iron chelation and reduction during early non-enzymatic wood decay processes have been studied in this thesis. The results indicate that the effects of the chelator/iron ratio, the pH, and other reaction parameters on the hydroxyl radical generation in a Fenton type system can be determined using ESR spin-trapping techniques. Data also support the hypothesis that superoxide radicals are involved in chelator-mediated Fenton processes. The mechanisms involved in free radical activation of Thermal Mechanical Pulp fibers were investigated. The activation of TMP fibers was evaluated by ESR measurement of free phenoxy radical generation on solid fibers. The results indicate that low molecular weight chelators can improve Fenton reactions, thus in turn stimulating the free radical activation of TMP fibers. A mediated Fenton system was evaluated for decolorization of several types of dyes. The result shows that the Fenton system mediated by a catecholate-type chelator effectively reduced the color of a diluted solution of synthetic dyes after 90 minutes of treatment at room temperature. The results show that compared to a neat Fenton process, the mediated Fenton decolorization process increased the production, and therefore the effective longevity, of hydroxyl radical species to increase the decolorization efficiency.

Sequential origin in the high performance properties of spider dragline silk

Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376 MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230 MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers.

Rapid prototyping of multi-scale biomedical microdevices by combining additive manufacturing technologies

The possibility of designing and manufacturing biomedical microdevices with multiple length-scale geometries can help to promote special interactions both with their environment and with surrounding biological systems. These interactions aim to enhance biocompatibility and overall performance by using biomimetic approaches. In this paper, we present a design and manufacturing procedure for obtaining multi-scale biomedical microsystems based on the combination of two additive manufacturing processes: a conventional laser writer to manufacture the overall device structure, and a direct-laser writer based on two-photon polymerization to yield finer details. The process excels for its versatility, accuracy and manufacturing speed and allows for the manufacture of microsystems and implants with overall sizes up to several millimeters and with details down to sub-micrometric structures. As an application example we have focused on manufacturing a biomedical microsystem to analyze the impact of microtextured surfaces on cell motility. This process yielded a relevant increase in precision and manufacturing speed when compared with more conventional rapid prototyping procedures.

Molecular recognition with nanostructures fabricated by photopolymerization within metallic subwavelength apertures

The first demonstration of fabrication of submicron lateral resolution molecularly imprinted polymer (MIP) patterns by photoinduced local polymerization within metal subwavelength apertures is reported. The size of the photopolymerized MIP features is finely tuned by the dose of 532 nm radiation. Rhodamine 123 (R123) has been selected as a fluorescent model template to prove the recognition capability of the MIP nanostructures, which has been evaluated by fluorescence lifetime imaging microscopy (FLIM) with single photon timing measurements. The binding selectivity provided by the imprinting effect has been confirmed in the presence of compounds structurally related to R123. These results pave the way to the development of nanomaterial architectures with biomimetic artificial recognition properties for environmental, clinical and food testing.

BR3: a biologically inspired fish-like robot actuated by SMA-based artificial muscles

Los peces son animales, donde en la mayoría de los casos, son considerados como nadadores muy eficientes y con una alta capacidad de maniobra. En general los peces se caracterizan por su capacidad de maniobra, locomoción silencioso, giros y partidas rápidas y viajes de larga distancia. Los estudios han identificado varios tipos de locomoción que los peces usan para generar maniobras y natación constante. A bajas velocidades la mayoría de los peces utilizan sus aletas pares y / o impares para su locomoción, que ofrecen una mayor maniobrabilidad y mejor eficiencia de propulsión. A altas velocidades la locomoción implica el cuerpo y / o aleta caudal porque esto puede lograr un mayor empuje y aceleración. Estas características pueden inspirar el diseo y fabricación de una piel muy flexible, una aleta caudal mórfica y una espina dorsal no articulada con una gran capacidad de maniobra. Esta tesis presenta el desarrollo de un novedoso pez robot bio-inspirado y biomimético llamado BR3, inspirado en la capacidad de maniobra y nado constante de los peces vertebrados. Inspirado por la morfología de los peces Micropterus salmoides o también conocido como lubina negra, el robot BR3 utiliza su fundamento biológico para desarrollar modelos y métodos matemáticos precisos que permiten imitar la locomoción de los peces reales. Los peces Largemouth Bass pueden lograr un nivel increíble de maniobrabilidad y eficacia de la propulsión mediante la combinación de los movimientos ondulatorios y aletas morficas. Para imitar la locomoción de los peces reales en una contraparte artificial se necesita del análisis de tecnologías de actuación alternativos, como arreglos de fibras musculares en lugar de servo actuadores o motores DC estándar, así como un material flexible que proporciona una estructura continua sin juntas. Las aleaciones con memoria de forma (SMAs) proveen la posibilidad de construir robots livianos, que no emiten ruido, sin motores, sin juntas y sin engranajes. Asi es como un pez robot submarino se ha desarrollado y cuyos movimientos son generados mediante SMAs. Estos actuadores son los adecuados para doblar la espina dorsal continua del pez robot, que a su vez provoca un cambio en la curvatura del cuerpo. Este tipo de arreglo estructural está inspirado en los músculos rojos del pescado, que son usados principalmente durante la natación constante para la flexión de una estructura flexible pero casi incompresible como lo es la espina dorsal de pescado. Del mismo modo la aleta caudal se basa en SMAs y se modifica para llevar a cabo el trabajo necesario. La estructura flexible proporciona empuje y permite que el BR3 nade. Por otro lado la aleta caudal mórfica proporciona movimientos de balanceo y guiada. Motivado por la versatilidad del BR3 para imitar todos los modos de natación (anguilliforme, carangiforme, subcarangiforme y tunniforme) se propone un controlador de doblado y velocidad. La ley de control de doblado y velocidad incorpora la información del ángulo de curvatura y de la frecuencia para producir el modo de natación deseado y a su vez controlar la velocidad de natación. Así mismo de acuerdo con el hecho biológico de la influencia de la forma de la aleta caudal en la maniobrabilidad durante la natación constante se propone un control de actitud. Esta novedoso robot pescado es el primero de su tipo en incorporar sólo SMAs para doblar una estructura flexible continua y sin juntas y engranajes para producir empuje e imitar todos los modos de natación, así como la aleta caudal que es capaz de cambiar su forma. Este novedoso diseo mecatrónico presenta un futuro muy prometedor para el diseo de vehículos submarinos capaces de modificar su forma y nadar mas eficientemente. La nueva metodología de control propuesto en esta tesis proporcionan una forma totalmente nueva de control de robots basados en SMAs, haciéndolos energéticamente más eficientes y la incorporación de una aleta caudal mórfica permite realizar maniobras más eficientemente. En su conjunto, el proyecto BR3 consta de cinco grandes etapas de desarrollo: • Estudio y análisis biológico del nado de los peces con el propósito de definir criterios de diseño y control. • Formulación de modelos matemáticos que describan la: i) cinemática del cuerpo, ii) dinámica, iii) hidrodinámica iv) análisis de los modos de vibración y v) actuación usando SMA. Estos modelos permiten estimar la influencia de modular la aleta caudal y el doblado del cuerpo en la producción de fuerzas de empuje y fuerzas de rotación necesarias en las maniobras y optimización del consumo de energía. • Diseño y fabricación de BR3: i) estructura esquelética de la columna vertebral y el cuerpo, ii) mecanismo de actuación basado en SMAs para el cuerpo y la aleta caudal, iii) piel artificial, iv) electrónica embebida y v) fusión sensorial. Está dirigido a desarrollar la plataforma de pez robot BR3 que permite probar los métodos propuestos. • Controlador de nado: compuesto por: i) control de las SMA (modulación de la forma de la aleta caudal y regulación de la actitud) y ii) control de nado continuo (modulación de la velocidad y doblado). Está dirigido a la formulación de los métodos de control adecuados que permiten la modulación adecuada de la aleta caudal y el cuerpo del BR3. • Experimentos: está dirigido a la cuantificación de los efectos de: i) la correcta modulación de la aleta caudal en la producción de rotación y su efecto hidrodinámico durante la maniobra, ii) doblado del cuerpo para la producción de empuje y iii) efecto de la flexibilidad de la piel en la habilidad para doblarse del BR3. También tiene como objetivo demostrar y validar la hipótesis de mejora en la eficiencia de la natación y las maniobras gracias a los nuevos métodos de control presentados en esta tesis. A lo largo del desarrollo de cada una de las cinco etapas, se irán presentando los retos, problemáticas y soluciones a abordar. Los experimentos en canales de agua estarán orientados a discutir y demostrar cómo la aleta caudal y el cuerpo pueden afectar considerablemente la dinámica / hidrodinámica de natación / maniobras y cómo tomar ventaja de la modulación de curvatura que la aleta caudal mórfica y el cuerpo permiten para cambiar correctamente la geometría de la aleta caudal y del cuerpo durante la natación constante y maniobras. ABSTRACT Fishes are animals where in most cases are considered as highly manoeuvrable and effortless swimmers. In general fishes are characterized for his manoeuvring skills, noiseless locomotion, rapid turning, fast starting and long distance cruising. Studies have identified several types of locomotion that fish use to generate maneuvering and steady swimming. At low speeds most fishes uses median and/or paired fins for its locomotion, offering greater maneuverability and better propulsive efficiency At high speeds the locomotion involves the body and/or caudal fin because this can achieve greater thrust and accelerations. This can inspire the design and fabrication of a highly deformable soft artificial skins, morphing caudal fins and non articulated backbone with a significant maneuverability capacity. This thesis presents the development of a novel bio-inspired and biomimetic fishlike robot (BR3) inspired by the maneuverability and steady swimming ability of ray-finned fishes (Actinopterygii, bony fishes). Inspired by the morphology of the Largemouth Bass fish, the BR3 uses its biological foundation to develop accurate mathematical models and methods allowing to mimic fish locomotion. The Largemouth Bass fishes can achieve an amazing level of maneuverability and propulsive efficiency by combining undulatory movements and morphing fins. To mimic the locomotion of the real fishes on an artificial counterpart needs the analysis of alternative actuation technologies more likely muscle fiber arrays instead of standard servomotor actuators as well as a bendable material that provides a continuous structure without joins. The Shape Memory Alloys (SMAs) provide the possibility of building lightweight, joint-less, noise-less, motor-less and gear-less robots. Thus a swimming underwater fish-like robot has been developed whose movements are generated using SMAs. These actuators are suitable for bending the continuous backbone of the fish, which in turn causes a change in the curvature of the body. This type of structural arrangement is inspired by fish red muscles, which are mainly recruited during steady swimming for the bending of a flexible but nearly incompressible structure such as the fishbone. Likewise the caudal fin is based on SMAs and is customized to provide the necessary work out. The bendable structure provides thrust and allows the BR3 to swim. On the other hand the morphing caudal fin provides roll and yaw movements. Motivated by the versatility of the BR3 to mimic all the swimming modes (anguilliform, caranguiform, subcaranguiform and thunniform) a bending-speed controller is proposed. The bending-speed control law incorporates bend angle and frequency information to produce desired swimming mode and swimming speed. Likewise according to the biological fact about the influence of caudal fin shape in the maneuverability during steady swimming an attitude control is proposed. This novel fish robot is the first of its kind to incorporate only SMAs to bend a flexible continuous structure without joints and gears to produce thrust and mimic all the swimming modes as well as the caudal fin to be morphing. This novel mechatronic design is a promising way to design more efficient swimming/morphing underwater vehicles. The novel control methodology proposed in this thesis provide a totally new way of controlling robots based on SMAs, making them more energy efficient and the incorporation of a morphing caudal fin allows to perform more efficient maneuvers. As a whole, the BR3 project consists of five major stages of development: • Study and analysis of biological fish swimming data reported in specialized literature aimed at defining design and control criteria. • Formulation of mathematical models for: i) body kinematics, ii) dynamics, iii) hydrodynamics, iv) free vibration analysis and v) SMA muscle-like actuation. It is aimed at modelling the e ects of modulating caudal fin and body bend into the production of thrust forces for swimming, rotational forces for maneuvering and energy consumption optimisation. • Bio-inspired design and fabrication of: i) skeletal structure of backbone and body, ii) SMA muscle-like mechanisms for the body and caudal fin, iii) the artificial skin, iv) electronics onboard and v) sensor fusion. It is aimed at developing the fish-like platform (BR3) that allows for testing the methods proposed. • The swimming controller: i) control of SMA-muscles (morphing-caudal fin modulation and attitude regulation) and ii) steady swimming control (bend modulation and speed modulation). It is aimed at formulating the proper control methods that allow for the proper modulation of BR3’s caudal fin and body. • Experiments: it is aimed at quantifying the effects of: i) properly caudal fin modulation into hydrodynamics and rotation production for maneuvering, ii) body bending into thrust generation and iii) skin flexibility into BR3 bending ability. It is also aimed at demonstrating and validating the hypothesis of improving swimming and maneuvering efficiency thanks to the novel control methods presented in this thesis. This thesis introduces the challenges and methods to address these stages. Waterchannel experiments will be oriented to discuss and demonstrate how the caudal fin and body can considerably affect the dynamics/hydrodynamics of swimming/maneuvering and how to take advantage of bend modulation that the morphing-caudal fin and body enable to properly change caudal fin and body’ geometry during steady swimming and maneuvering.

Evaluation of the surface free energy of plant surfaces: toward standardizing the procedure

Plant surfaces have been found to have a major chemical and physical heterogeneity and play a key protecting role against multiple stress factors. During the last decade, there is a raising interest in examining plant surface properties for the development of biomimetic materials. Contact angle measurement of different liquids is a common tool for characterizing synthetic materials, which is just beginning to be applied to plant surfaces. However, some studies performed with polymers and other materials showed that for the same surface, different surface free energy values may be obtained depending on the number and nature of the test liquids analyzed, materials' properties, and surface free energy calculation methods employed. For 3 rough and 3 rather smooth plant materials, we calculated their surface free energy using 2 or 3 test liquids and 3 different calculation methods. Regardless of the degree of surface roughness, the methods based on 2 test liquids often led to the under- or over-estimation of surface free energies as compared to the results derived from the 3-Liquids method. Given the major chemical and structural diversity of plant surfaces, it is concluded that 3 different liquids must be considered for characterizing materials of unknown physico-chemical properties, which may significantly differ in terms of polar and dispersive interactions. Since there are just few surface free energy data of plant surfaces with the aim of standardizing the calculation procedure and interpretation of the results among for instance, different species, organs, or phenological states, we suggest the use of 3 liquids and the mean surface tension values provided in this study.

Caracterización de las Propiedades Dinámicas de la Seda de Araña

El término Biomimética se ha hecho común en los medios científicos, se refiere al trabajo de diversos científicos (ingenieros, químicos, físicos, biólogos, etc.) que tratan de copiar los procesos biológicos y aplicarlos en distintas áreas tecnológicas y científicas. En este campo científico, uno de los productos naturales que llama más la atención es la telaraña. Numerosos científicos en todo el mundo tratan de copiar las propiedades de la seda que produce la araña, y lo más interesante es que hasta intentan reproducir el método que usan las arañas para fabricar la seda De la bibliografía consultada, se desprende la importancia de la seda en la vida de las arañas, pues toda actividad que realizan tiene que ver de alguna manera con este elemento. Uno de estos elementos es la tela de araña orbicular, que representa el objeto principal para la supervivencia de la araña y su especie. Las investigaciones realizadas en esta línea nos proporcionan información sobre sus magníficas propiedades mecánicas como de resistencia, elasticidad y tenacidad del hilo de seguridad (seda MA) segregada por una araña de la especie Argiope Argentata. El enfoque de la presente tesis se realiza desde una perspectiva analítica-experimental, tomando a la tela de araña como una clase especial de sistemas pretensados, llamados Tensegrity Structures. Se desarrolla un modelo conceptual que describe en forma aproximada el comportamiento dinámico de una estructura hecha de seda MA. Haciendo uso de las técnicas experimentales de vibraciones libres se realizan los ensayos experimentales. La evaluación de los resultados analíticos y experimentales reflejan claramente que la función principal de la tela de araña es la de convertir energía cinética en energía de deformación y primordialmente en energía de disipación, el cual se efectúa gracias a las propiedades viscoelásticas de la seda. La araña en forma instintiva recurre a la ayuda del aire (como elemento disipador) para el buen funcionamiento de la tela de araña al momento de la captura de las presas, disipándose el 99% de la energía total en los tres primeros ciclos de oscilación de la tela de araña luego del impacto de la presa. ABSTRACT The term Biomimetic has become a very common word in the scientific world to describe the reproduction of the biological processes and its application in the different technogycal and scientific areas. One of the most notably natural product of this field is the Spiderweb. In the present days, many scientists of the world are active working in the reproduction of the proprieties of the Spiderweb. Most interesting even more, is the attempt to reproduce the process of the production of the Spiderweb by the spider. Most of the bibliography references deals whit the importance of the Spiderweb silk in the life of the spiders and the orbicular Spiderweb represents the spider survival and of the species. The research conducted in this field provide information about the excellent mechanical proprieties such us strength, elasticity and tenacity of the safety fiber (silk MA Drag-line) segregated by a spider of the Argiope Argentata species. The present work is oriented to an analytical and experimental study considering the Spiderweb as a special class of pre-stressed systems called Tensegrity (tensional integrity) structures. A conceptual model maked up by a cord und a point mass has been developed. This model approximates the dynamics performance of the structure made of the silk MA. The evaluation of the analytical and experimental results clear described that the main function of the Spiderweb is the transformation of the kinetic energy in deformation energy, and mainly in dissipation energy thank to the viscoelastic proprieties of the Spiderweb. With the help of the Spiderweb, the spider instinctively resorts to the help of the surroundings air as a dissipation element. This permits to dissipative the 99% of the total energy during the three first oscillations cycles of the web after the impact of the victim.

Sequence-specific polypeptoids: A diverse family of heteropolymers with stable secondary structure

We have synthesized and characterized a family of structured oligo-N-substituted-glycines (peptoids) up to 36 residues in length by using an efficient solid-phase protocol to incorporate chemically diverse side chains in a sequence-specific fashion. We investigated polypeptoids containing side chains with a chiral center adjacent to the main chain nitrogen. Some of these sequences have stable secondary structure, despite the achirality of the polymer backbone and its lack of hydrogen bond donors. In both aqueous and organic solvents, peptoid oligomers as short as five residues give rise to CD spectra that strongly resemble those of peptide α-helices. Differential scanning calorimetry and CD measurements show that polypeptoid secondary structure is highly stable and that unfolding is reversible and cooperative. Thermodynamic parameters obtained for unfolding are similar to those obtained for the α-helix to coil transitions of peptides. This class of biomimetic polymers may enable the design of self-assembling macromolecules with novel structures and functions.

Isolation and one-step preparation of A2E and iso-A2E, fluorophores from human retinal pigment epithelium

Age-related macular degeneration, a major cause of blindness for which no satisfactory treatments exist, leads to a gradual decrease in central high acuity vision. The accumulation of fluorescent materials, called lipofuscin, in retinal pigment epithelial cells of the aging retina is most pronounced in the macula. One of the fluorophores of retinal pigment epithelial lipofuscin has been characterized as A2E, a pyridinium bis-retinoid, which is derived from two molecules of vitamin A aldehyde and one molecule of ethanolamine. An investigation aimed at optimizing the in vitro synthesis of A2E has resulted in the one-step biomimetic preparation of this pigment in 49% yield, readily producing more than 50 mg in one step. These results have allowed for the optimization of HPLC conditions so that nanogram quantities of A2E can be detected from extracts of tissue samples. By using 5% of the extract from individual aged human eyes, this protocol has led to the quantification of A2E and the characterization of iso-A2E, a new A2E double bond isomer; all-trans-retinol and 13-cis-retinol also have been identified in these HPLC chromatograms. Exposure of either A2E or iso-A2E to light gives rise to 4:1 A2E:iso-A2E equilibrium mixtures, similar to the composition of these two pigments in eye extracts. A2E and iso-A2E may exhibit surfactant properties arising from their unique wedge-shaped structures.

Study of interaction between Si(O,C) nanowires and the biological system

Nanomedicine is a new branch of medicine, based on the potentiality and intrinsic properties of nanomaterials. Indeed, the nanomaterials ( i.e. the materials with nano and under micron size) can be suitable to different applications in biomedicine. The nanostructures can be used by taking advantage of their properties (for example superparamagnetic nanoparticles) or functionalized to deliver the drug in a specific target, thanks the ability to cross biological barriers. The size and the shape of 1D-nanostructures (nanotubes and nanowires) have an important role on the cell fate: their morphology plays a key role on the interaction between nanostructure and the biological system. For this reason the 1D nanostructure are interesting for their ability to mime the biological system. An implantable material or device must therefore integrate with the surrounding extracellular matrix (ECM), a complex network of proteins with structural and signaling properties. Innovative techniques allow the generation of complex surface patterns that can resemble the structure of the ECM, such as 1D nanostructures. NWs based on cubic silicon carbide (3C-SiC), either bare (3C-SiC NWs) or surrounded by an amorphous shell (3C-SiC/SiO2 core/shell NWs), and silicon oxycarbide nanowires (SiOxCy NWs) can meet the chemical, mechanical and electrical requirements for tissue engineering and have a strong potential to pave the way for the development of a novel generation of implantable nano-devices. Silicon oxycarbide shows promising physical and chemical properties as elastic modulus, bending strength and hardness, chemical durability superior to conventional silicate glasses in aggressive environments and high temperature stability up to 1300 °C. Moreover, it can easily be engineered through functionalization and decoration with macro-molecules and nanoparticles. Silicon carbide has been extensively studied for applications in harsh conditions, as chemical environment, high electric field and high and low temperature, owing to its high hardness, high thermal conductivity, chemical inertness and high electron mobility. Also, its cubic polytype (3C) is highly biocompatible and hemocompatible, and some prototypes of biomedical applications and biomedical devices have been already realized starting from 3C-SiC thin films. Cubic SiC-based NWs can be used as a biomimetic biomaterial, providing a robust and novel biocompatible biological interface . We cultured in vitro A549 human lung adenocarcinoma epithelial cells and L929 murine fibroblast cells over core/shell SiC/SiO2, SiOxCy and bare 3C-SiC nanowire platforms, and analysed the cytotoxicity, by indirect and direct contact tests, the cell adhesion, and the cell proliferation. These studies showed that all the nanowires are biocompatible according to ISO 10993 standards. We evaluated the blood compatibility through the interaction of the nanowires with platelet rich plasma. The adhesion and activation of platelets on the nanowire bundles, assessed via SEM imaging and soluble P-selectin quantification, indicated that a higher platelet activation is induced by the core/shell structures compared to the bare ones. Further, platelet activation is higher with 3C-SiC/SiO2 NWs and SiOxCyNWs, which therefore appear suitable in view of possible tissue regeneration. On the contrary, bare 3C-SiC NWs show a lower platelet activation and are therefore promising in view of implantable bioelectronics devices, as cardiovascular implantable devices. The NWs properties are suitable to allow the design of a novel subretinal Micro Device (MD). This devices is based on Si NWs and PEDOT:PSS, though the well know principle of the hybrid ordered bulk heterojunction (OBHJ). The aim is to develop a device based on a well-established photovoltaic technology and to adapt this know-how to the prosthetic field. The hybrid OBHJ allows to form a radial p–n junction on a nanowire/organic structure. In addition, the nanowires increase the light absorption by means of light scattering effects: a nanowires based p-n junction increases the light absorption up to the 80%, as previously demonstrated, overcoming the Shockley-Queisser limit of 30 % of a bulk p-n junction. Another interesting employment of these NWs is to design of a SiC based epicardial-interacting patch based on teflon that include SiC nanowires. . Such contact patch can bridge the electric conduction across the cardiac infarct as nanowires can ‘sense’ the direction of the wavefront propagation on the survival cardiac tissue and transmit it to the downstream surivived regions without discontinuity. The SiC NWs are tested in terms of toxicology, biocompatibility and conductance among cardiomyocytes and myofibroblasts.

Atividade da própolis verde contra o fitopatógeno Pythium aphanidermatum e análise da interação do composto majoritário Artepillin C com sistemas biomiméticos de membranas

O aumento da resistência microbiana devido a fatores como uso excessivo e ineficiente de antibióticos convencionais acarreta a necessidade da busca por novos compostos bioativos que atuem por mecanismos de ação diferentes aos fármacos já conhecidos. Na agricultura, o uso intensivo de pesticidas para o combate de microrganismos que comprometem principalmente a parte alimentícia também traz diversos problemas relacionados à resistência antimicrobiana e a riscos ambientais, oriundos do acúmulo dessas substâncias no solo. Dentro deste aspecto, o pseudofungo Pythium aphanidermatum, da classe dos oomicetos, destaca-se por ser uma espécie agressiva e altamente resistente a fungicidas comuns, apodrecendo raízes e frutos de cultivos de tomate, beterraba, pepino, pimentão, etc. A própolis verde, constituída em sua grande parte por material resinoso coletado e processado pela abelha da espécie Apis mellifera tem sido utilizada na medicina tradicional devido ao seu amplo espectro de ações preventivas e tratamentos de doenças, possuindo propriedades anti-inflamatórias, antimicrobianas, anticancerígenas e antioxidantes, tornando-se um produto de grande interesse na busca de novos compostos bioativos. Dentro destes aspectos apresentados, neste trabalho investigamos a ação da própolis verde contra o fitopatógeno P. aphanidermatum e identificamos através da técnica de cromatografia e bioensaios que a Artepillin C (3,5-diprenil-4-ácido-hidroxicinâmico), majoritária na própolis verde, foi o principal composto nesta ação. Os efeitos terapêuticos desta molécula tem sido foco de muitos estudos, porém ainda não há evidência em sua interação com agregados anfifílicos que mimetizam membranas celulares. O caráter anfifílico do composto, elevado pela presença dos grupos prenilados ligados ao ácido cinâmico, favoreceram a sua inserção nas membranas modelo, principalmente em seu estado agregado. Estas conclusões puderam ser inferidas devido às alterações nas propriedades das bicamadas lipídicas na presença da Artepillin C, podendo causar, especificamente para o caso de fitopatógenos como o P. aphanidermatum, perdas funcionais das proteínas de membranas, liberação de eletrólitos intracelulares e desintegração citoplasmática dos micélios e esporos. Ainda, as diferentes composições lipídicas nas vesículas influenciam no modo de interação do composto e consequentes alterações em suas estruturas, principalmente na presença do colesterol, que auxilia na manutenção da permeabilidade da bicamada lipídica, que pode contribuir para a integridade do conteúdo citoplasmático da célula.