Chun-Long Chen, Pacific Northwest National Laboratory
Nico Sommerdijk, Eindhoven University of Technology
Tiffany Walsh, Deakin University
Shuguang Zhang, Massachusetts Institute of Technology
Pacific Northwest National Laboratory
BM09.01: Biomimetic Materials Based on Peptide Self-Assembly
Monday PM, November 26, 2018
Sheraton, 2nd Floor, Back Bay A
8:00 AM - BM09.01.01
Self-Organization of Peptides in Bioinspired Vesicles—Role of Relative Concentration and Helical Separation
Akash Banerjee1,Meenakshi Dutt1
Rutgers University1Show Abstract
Biological cells can inspire the creation of nanoparticles equipped to store and release hydrophobic drug molecules upon demand. Lipid vesicles impregnated with alpha-helical peptides have demonstrated the clustering of the peptides under equilibrium. The formation of thick, amphiphilic, transmembrane channels via the self-organization of the peptides could be potentially used for the on-demand release of small drug molecules from the hydrophobic core of a vesicle bilayer. We are interested in understanding the driving forces responsible for cluster formations and evaluating their effects using the Molecular Dynamics simulation technique. Coarse grained representations of the molecules are used to resolve the extended spatiotemporal scales relevant to the problem at hand. The bonded and non-bonded interactions between the particles is captured by the Martini force field. We investigate the role of peptide concentration and helical separation on the cluster formation. We find the cluster size to be dependent more on helical separation as compared to peptide concentration. Additionally, we test the role of hydrophobic mismatch to understand the effect of electrostatic interactions between the peptides and lipid molecules. Our results demonstrate negative mismatch to result in larger cluster sizes as compared to a zero hydrophobic mismatch condition due to larger perturbations in the vesicle monolayers.
8:15 AM - BM09.01.02
Self-Assembly of Membrane-Active Peptides into Macromolecular-Size Pores
Kalina Hristova1,Sijia Li1,Sarah Kim1,Anna Pittman2,Gavin King2,William Wimley3
Johns Hopkins University1,University of Missouri2,Tulane University3Show Abstract
Peptides that self-assemble into pore-like structures in lipid bilayers could have utility in a variety of biotechnological and clinical applications due to their ability to breach the barrier imposed by lipid bilayers. To empower such discoveries, we use rationally designed peptide libraries and high-throughput screens to select peptides based on a particular property, in this case macromolecular-size bilayer poration. Towards this goal, we designed a library based on the bee venom peptide melittin, and we developed a high throughput screen that reports on the passage of macromolecules across lipid bilayers. We identified two peptide families that efficiently assemble into large pore-like structures. One of the families is highly active at pH 7. The other peptide family is pH sensitive, as its self-assembly is triggered by low pH. The pH-triggered peptides could be used for endosomal release of uptaken polar molecules into the cell cytosol, upon endosomal acidification. They also could be used in cancer therapies to selectively permeabilize the plasma membranes of cancer cells, since the vicinity of solid tumors is often acidic. Additional generations can be screened to further fine-tune the properties of these peptides.
8:30 AM - BM09.01.03
Exploring the Tubability of the Aggregation and Gelation Process of the Tripeptide Glycyl-Alanyl-Glycine (GAG)
David DiGuiseppi1,Lavenia Thursch1,Nicolas Alvarez1,Reinhard Schweitzer-Stenner1
Drexel University1Show Abstract
Self-assembly of biomolecules is a prominent issue explored in biomedical, biophysical, and bio-material research. Understanding how and why certain peptides/proteins prefer to self-assemble into larger networks can reveal the mechanism of amyloid formation and assist in bottom-up designs of supramolecular structures like gels and nanotubes. Some low molecular weight di- or tripeptides with aromatic residues and terminal groups have been shown to form gels. Contrary to expectations, we recently discovered that cationic glycylalanylglycine (GAG), a tripeptide of low hydrophobicity, forms a gel in 55 mol% ethanol/45 mol% water at room temperature if the concentration exceeds 200 mM. The underlying structure is comprised of unusually long crystalline fibrils (in the 10-5m range), which do not exhibit the canonical β-sheet structure. Rheological data and vibrational circular dichroism spectra suggest the existence of two different gel phases, one formed between 15° and 35°C with left handed twisted fibrils and G’ values at ca. 2*104 Pa and another one formed below 15°C with right handed twisted fibrils and G’ values close to 105 Pa. Results from DFT calculations indicate that the two phases might be underlied by rather differently structured fibrils. The fluorescence kinetics probing the incorporation of thioflavin T into the hydrophobic interior of fibrils indicate a retarded diffusion of the fluorophore into fibrils that formed rather quickly after incubation above 15°C, while fluorescence increase, and gelation proceed on a similar time scale for the gel phase formed below this temperature. Upon increasing the temperature, it can preserve this capability until the melting temperature is reached, which suggests that this gel phase has all what it takes to function as a drug delivery system. The potential reformation process of the fibrils probed by UVCD, rheology, and microscopy show that after sitting for 16h above the melting temperature, the fibrils do not have the ability to grow back. Instead, microscopic images suggest the formation of a crystal-type structure that forms in its place. Our results therefore suggest that the gel phases are meta-stable states of the system that form more quickly at or below room temperature. We care currently working on optimizing the gelation/melting conditions for specific biotechnological applications of the gel as well as characterizing the observed crystal-type structure.
8:45 AM - BM09.01.04
Neutral Self-Assembling Multidomain Peptides—Steric Impediment Regulates Nanofiber Formation and Materials Properties
Tania Lopez Silva1,David Leach1,I-Che Li1,Xinran Wang1,Jeffrey Hartgerink1
Rice University1Show Abstract
Peptide-based materials have drawn high interest for their use in biomedical applications such as drug delivery, cell encapsulation, and tissue regeneration. Particularly, self-assembling peptide hydrogels have shown promising properties as biomaterials since their properties and functionality are tunable by their peptide sequence. For example, they are inherently biocompatible and biodegradable, their nanofibrous structure resembles the extracellular matrix, and they form materials with high-water content. Generally, these peptides utilize ionic amino acids to control self-assembly by changing the pH or ionic strength. Included in these group are the self-assembling Multidomain Peptide nanofibers (MDP), composed of an amphiphilic β-sheet forming core and flanking charged domains, which increase peptide solubility and make the peptide material responsive to pH changes and the presence of ions.
It is known that the biological response and cell behavior is highly dependent on the chemistry of the materials. Positive polymers promote cell adhesion and proliferation while showing concentration-dependent cytotoxicity, whereas neutral polymers, such as PEG, are frequently inert, biocompatible and non-immunogenic. Previously, all MDPs were either positively or negatively charged; therefore, expanding the scope of MDPs to neutral, non-ionic peptides will make distinct biological properties available that are not present in highly charged peptides.
Strategies to control the self-assembly of non-ionic peptides is limited because these peptides tend to have low solubility, aggregate or precipitate in aqueous solutions, making the formation of finite supramolecular structures and self-assembled hydrogels challenging. In this project, we present an alternative mechanism to control the self-assembly of neutral, uncharged multidomain peptides by utilizing steric impediment. Through the study of a series of neutral peptides, we analyzed the effect of the steric interactions on the peptide solubility, aggregation, nanostructure, and hydrogelation. From the series, a novel neutral multidomain peptide hydrogel was developed, which is inert to pH variation and ionic strength. This novel material showed promising properties for biomedical, cell preservation and tissue regeneration applications.
9:00 AM - *BM09.01.05
Self-Assembly of 2D Peptide-Based Crystalline Nanomaterials
Emory University1Show Abstract
Structurally defined materials on the nanometer length-scale have been historically the most challenging to rationally construct and the most difficult to structurally analyze. Sequence-specific biomolecules, i.e., proteins and nucleic acids, have advantages as design elements for construction of these types of nano-scale materials in that correlations can be drawn between sequence and higher order structure, potentially affording ordered assemblies in which functional properties can be controlled through the progression of structural hierarchy encoded at the molecular level. The predictable design of self-assembled structures requires precise structural control of the interfaces between peptide subunits (protomers). However, control of quaternary structure has proven to be challenging to reliably predict, as conservative changes in sequence can result in significant changes in higher order, i.e., supramolecular, structure. We have employed simple self-assembling peptides as building blocks for the construction of two-dimensional nano-scale assemblies. In contrast to filamentous assemblies (e.g., fibrils, ribbons, and tubes), protein-based two-dimensional assemblies occur relatively infrequently in native biological systems. We have demonstrated that extended and structurally defined two-dimensional assemblies can be constructed through lateral association of chiral rod-like subunits such as the collagen triple helix. The resultant assemblies can exhibit sequence-dependent control of structure, including growth in the lateral and/or axial dimensions. Moreover, the sheet-like assemblies can be integrated with other self-assembled biological structural motifs, such as DNA origami nano-tiles, to afford self-organized hybrid assemblies. Despite the potential for these two-dimensional assemblies as structurally defined nano-scale scaffolds, it remains challenging to reliably predict and control the structure of the assemblies based on sequence-structure correlations at present.
10:00 AM - *BM09.01.06
Bio-Inspired Materials Linking Covalent and Supramolecular Polymers
Northwestern University1Show Abstract
Supramolecular soft matter is a rapidly emerging field that encompasses the rational use of organic molecules to design function in materials. The most promising systems are “supramolecular polymers” since one-dimensional catenation of structural units is a critical feature to create mechanically robust macroscopic systems and directed transport of charge in aligned morphologies. Supramolecular polymers, in contrast to macromolecules in which structural units are linked through covalent bonds, supramolecular systems are designed using additive noncovalent bonds that are tunable over a very broad range of binding energies encoded in the molecular structure of the “mers”. Furthermore, a major gap in the design of synthetic soft matter is the rational integration of covalent and supramolecular polymers, a concept that is used to craft function in the structures of living organisms. This lecture will describe first entirely supramolecular systems based on peptides and nucleic acids in which dynamics of non-covalently bonded monomers can reversibly form superstructures linked to mechanical and biological functions. Within the domain of hybrid systems in which covalent macromolecules are integrated with supramolecular structures, the lecture will describe materials inspired by muscles that are capable of transducing thermal to mechanical energy, light to mechanical energy, and light to chemical energy in photocatalytic materials.
10:30 AM - BM09.01.07
Self-Assembly of Hierarchical Cellular Materials from Amphiphic Triblock Peptides
Erik Spoerke1,Brad Jones1,Jill Wheeler1,Jeffrey Vervacke1,Christina Ting1,Mark Stevens1
Sandia National Laboratories1Show Abstract
Macromolecular self-assembly in biological systems takes many forms and enables countless functions across multiple length scales. Often, the structure and function of these assembled structures are dictated by subtle changes in the composition of the molecular building blocks that make up these materials. For example, simple amino acid substitutions can impart significant changes in the structure and function of protein assemblies. Inspired by this theme, we explore here the self-assembly of an ABC triblock peptide-oligoethylene oxide amphiphile with hydrophilic A and C blocks and a hydrophobic B peptide block. By varying the amino acid side chain size and hydrophobicity within the B-block, we observe aqueous self-assembly into polymorphic cellular particles with hierarchical structure and porosity ranging from giant vesicles with foam-like membranes to porous tubular architectures. These structures are characterized microscopically and spectroscopically to determine the relationships between the varied peptide compositions, tunable intermolecular interactions, and the observed morphologies. Additional evaluation of these materials as vehicles for molecular encapsulation and as templates for secondary mineral templating reveal potential new strategies to control hierarchical materials synthesis and assembly through bio-inspired molecular building block design.
Sandia National Laboratories is a multi-mission laboratory managed and operated by National Technology and Engineering Solutions of Sandia, LLC., a wholly owned subsidiary of Honeywell International, Inc., for the U.S. Department of Energy’s National Nuclear Security Administration under contract DE-NA0003525.
10:45 AM - BM09.01.08
Modular Peptide-Polymer Conjugates—A Platform Technology for Mucin Analogues
Daniel French1,Luis Navarro1,Stefan Zauscher1
Duke Univ1Show Abstract
Mucins – the glycoprotein building-blocks of mucus – play diverse and crucial roles in the body. These functions range from lubrication of articular joints and the eye, to the protection of stomach endothelium from the harsh environment of the lumen, to modulation of microflora populations in the digestive and respiratory systems. Despite this diversity, these functions are all attributed to slight modifications in a general structure shared by all mucins: a telechelic triblock polypeptide comprised of terminal association moieties and a heavily glycosylated core which forms a hydrated bottle brush center. In vivo, these versatile functions are achieved by altering glycosylation patterns, crosslinking density, and targeting affinity in a modular fashion.
Inspired by this adaptability, we have emulated this general architecture in a modular conjugate analogue mucin platform which engenders general structural features preserved among mucins which we, and others, have identified as key to their function. To recapitulate the mucin backbone, we genetically tether and co-express terminal binding modules with a lysine-rich, elastin-like polypeptide (ELP) central scaffold. Binding modules may include sequences designed to target surfaces of interest, to facilitate intramolecular associations, or to direct surface conformation of our construct. The regularly-spaced lysines in the ELP scaffold can be harnessed for grafting synthetic polymer bristles. Bristle chemistry may be chosen for a desired property (including non-fouling character and lubricity) independent of the binding and scaffold modules. Our platform is, to our knowledge, the first to adapt the modularity of the mucin architecture into a bio-synthetic platform technology.
To demonstrate the application of our platform to clinically-relevant problems, we have tailored our mCAMP to osteoarthritis and kidney stone disease, two conditions infamous for profound morbidity and high prevalence. In tailoring our analogue mucin to cartilage, we hope to rival the performance of lubricin, a natural mucin which provides lubrication and wear protection to articular joints. Moreover, we seek to harness the properties of natural mucins and apply them to systems not naturally protected by mucinous coatings. In doing so, we have adapted our platform to binding calcium oxalate kidney stones. Association modules are designed to direct assembly on mineral surfaces as well as inhibit further mineralization. Moreover, these modules are designed to form intramolecular associations, facilitating a robust surface coating. The inclusion of non-fouling synthetic polymer bristles provides a means by which to inhibit protein-mediated crystal aggregation. In this platform technology, we have begun to develop a means by which to replicate not only the in vivo function of mucins, but to harness that function to meet additional clinical needs.
11:00 AM - BM09.01.09
Design of Bioresponsive Nanogels Inspired by Peptide-Glycan Interactions
Andrew Simonson1,Atip Lawanprasert1,Tyler Goralski1,Kenneth Keiler1,Scott Medina1
The Pennsylvania State University1Show Abstract
Early investigations from The Medina Group identified that binding of cationic membrane-active peptides with negatively charged cell-surface glycans was a critical initiating step to potentiate the peptide’s lytic action. Inspired by this natural system, we have designed a family of biohybrid nanomaterials assembled via electrostatic association of cationic peptides and anionic carbohydrates. Screening a series of peptide-polysaccharide pairs under electrospray synthesis conditions identified that poly-L-lysine (PLL) and hyaluronic acid (HA) rapidly co-assemble to yield nano-scale gel-like particles, which we refer to as nanogels. Importantly, using peptide-carbohydrate co-assembly allowed for direct encapsulation of both small molecule drugs and protein cargo into the nanogel carrier under mild aqueous conditions conducive to sensitive biomolecules. Further, we found that modulating the ratio of PLL and HA utilized during particle assembly yielded nanogels with tunable swelling profiles and failure rates, thus allowing for controlled temporal release of loaded cargo. In vitro testing demonstrates that nanogels exhibit versatile and complimentary mechanisms of cargo delivery depending on the biologic context. In mammalian cells, nanogels can deliver membrane-impermeable protein cargo to the cytoplasm by rapid internalization via endocytosis, followed by endosomal escape. Likewise, chemotherapeutic-loaded nanogels were capable of enhancing the potency of loaded drug by up to an order of magnitude towards both chemo-sensitive and -resistant tumor cell lines. Remarkably, in the presence of bacterial pathogens, nanogels show a very different behavior. The carrier is able to recruit microbes and permeabilize their cell wall to sensitize pathogens to the action of loaded antibiotic. In a notable example, delivery of vancomycin from nanogels enhanced the drug’s potency by >15-fold towards a gram-positive strain. Even more surprising was the ability of nanogels to sensitize gram-negative pathogens to the action of vancomycin, which are otherwise innately resistant to the drug due to the low permeability of their cell wall. This adaptable bioactivity, in combination with their low toxicity towards human endothelial cells and erythrocytes, demonstrate that nanogels represent a versatile and bio-responsive carrier capable of augmenting and enhancing the utility of a broad range of biomolecular cargoes.
11:15 AM - BM09.01.10
Template-Driven Peptide Assembly Yields Ultrasound Guided Phase-Changing Nanomaterials
Janna Sloand1,Scott Zinck1,Joel Schneider2,Julianna Simon1,Scott Medina1
The Pennsylvania State University1,National Institutes of Health2Show Abstract
Phase-changing nanoparticles (PCNs) are a class of materials that undergo solid-liquid-gas transitions in response to various engineered stimuli, leading to their application in fields that include thermal energy storage, bioelectronics and precision medicine. In particular, liquid-shelled perfluorocarbon PCNs that can be vaporized upon exposure to ultrasound (US) are poised to open unprecedented opportunities in nanomedicine and molecular imaging. However, despite recent progress, challenges remain in controlling the material properties and acoustic activation of PCNs, as well as overcoming the poor loading efficiency and delivery of biologic cargo from the carrier. Here, we describe the design and synthesis of a new class of PCNs recently prepared via templated peptide assembly, which we refer to as ‘nano-peptisomes’. Nano-peptisome architecture develops from the spontaneous orientation of de novo designed peptide amphiphiles around an US-sensitive fluorinated droplet as the template. Utilizing peptide-assembly allows for facile particle synthesis, direct incorporation of bioactive sequences displayed from the peptide corona, and the ability to easily encapsulate biologics during particle preparation using a mild solvent exchange procedure. We find that nano-peptisome size can be precisely controlled by simply modulating the starting peptide and fluorinated solvent concentrations during synthesis, leading to programmable acoustic properties of the final carrier. Further, biomolecular cargo, including peptides and proteins, can be encapsulated within the particle core and directly delivered to the cytoplasm of cells upon US-mediated rupture of the carrier. Bio-imaging studies demonstrate that nano-peptisomes can be tracked and guided using diagnostic B-mode US, while Doppler imaging allows for real-time monitoring of particle activation and rupture in tissue mimetic gels. These results establish nanopeptisomes as a novel theranostic platform capable of image-guided delivery of bioactive macromolecules into cells with spatial and temporal precision.
11:30 AM - *BM09.01.11
Biomolecules for Non-Biological Things—Materials Construction Through Peptide Design and Solution Assembly
University of Delaware1Show Abstract
Self-assembly of molecules is an attractive materials construction strategy due to its simplicity in application. By considering peptidic molecules in the bottom-up materials self-assembly design process, one can take advantage of inherently biomolecular attributes; intramolecular folding events, secondary structure, and electrostatic interactions; in addition to more traditional self-assembling molecular attributes such as amphiphilicty, to define hierarchical material structure and consequent properties. A new solution assembled system comprised of theoretically designed coiled coil bundle motifs will be introduced. The molecules and nanostructures are not natural sequences and provide opportunity for arbitrary nanostructure creation with peptides. With control of the display of all amino acid side chains (both natural and non-natural) throughout the peptide bundles, desired physical and covalent (through appropriate “click” chemistry) interactions have been designed to produce one and two-dimensional nanostructures. One-dimensional nanostructures span exotically rigid rod molecules that produce a wide variety of liquid crystal phases to semi-flexible chains, the flexibility of which are controlled by the interbundle linking chemistry. The two dimensional nanostructure is formed by physical interactions and are nanostructures not observed in nature. All of the assemblies are responsive to temperature since the individual bundle building blocks are physically stabilized coiled coil bundles that can be melted and reformed with temperature. Additional, novel nanostructures to be discussed include uniform nanotubes as well as the templated growth of metallic nanoparticle on and in peptide nanostructures. Included in the discussion will be molecule design, hierarchical assembly pathway design and control, click chemistry reactions, and the characterization of nanostructure as well as inherent material properties (e.g. extreme stiffness, responsiveness to temperature and pH, stability in aqueous and organic solvents).
BM09.02: Peptide-Based Nanomaterials
Monday PM, November 26, 2018
Sheraton, 2nd Floor, Back Bay A
1:30 PM - *BM09.02.01
Guiding Principles for Peptide-Based, Life-Like Nanotechnology
Hunter College1Show Abstract
Life’s diverse molecular functions are largely based on only a small number of highly conserved building blocks- the twenty canonical amino acids. These building blocks are chemically simple, but when they are organized in three-dimensional structures of tremendous complexity, new properties emerge, giving rise to the extraordinary machinery of life. So, if just twenty simple building blocks- when appropriately assembled – give rise to the complexity and functionality that can sustain life- then this is clearly a very versatile construction set. Our overall goal is conceptually simple: to figure out how to make nanoscale systems and materials from biology’s building blocks, and to apply these materials to diverse problems, that require them to be interfaced, ideally seamlessly, with living systems, or the natural environment. Different from other research groups, we have an unbiased approach, that is not guided by copying biological systems, and we keep these systems as simple as possible, which lowers barriers to application. The talk will focus on our latest results in three areas: (i) directed discovery of peptide nanostructures with new functions, by searching the sequence space; (ii) application of peptide nanostructures as functional materials (including customizable melanin pigments). (iii) actively assembling systems, that continuously turn over chemical fuels, enabling dynamic changes in structure and function.
2:00 PM - BM09.02.02
Large-Scale Self-Sorting in Supramolecular Assemblies
Charlotte Chen1,Liam Palmer1,2,Samuel Stupp1,2
Northwestern University1,Simpson Querrey Institute for BioNanotechnology2Show Abstract
Hierarchical organization across length scales is ubiquitous in the superstructures of living organisms. These highly functional structures form through self-assembly, and have therefore inspired significant research activity on synthetic supramolecular materials over the past decade. We report here on a synthetic system containing two supramolecular nanoscale polymers, of very similar structure, that interestingly exhibit micron scale self-sorting. The two different supramolecular polymers are formed by peptide amphiphiles and each is labeled with a different small fluorescent dye, and based on earlier work were expected to undergo molecular exchange. We hypothesize that electrostatic charges on the nanofibers promote the self-organization of the fibers into micron scale hierarchical structures, which take the form of 2D crystals, in order to minimize charge repulsion. The propensity to self-sort is diminished when ions are present to screen the electrostatic charges thus disrupting the hierarchical structures. Our results provide insight on strategies to promote self-sorting superstructures versus co-assembly in supramolecular systems.
2:15 PM - BM09.02.03
Thermally Reversible Transmembrane Molecular Channels Formed by Self-Assembled Metal-Organic Complexes
King Abdullah University of Science and Technology1Show Abstract
Biological channels are molecular gatekeepers that control cellular traffic across cell membrane. Realizing the functional principle of these systems through artificial transmembrane pores with molecularly defined structures is instrumental for future bionanotechnology applications. In this work, thermoresponsive synthetic channels based on supramolecular metal-organic complexes (MOCs) have been constructed to transport cell impermeable cargo across the membrane. The channels can be reversibly controlled as they collapse when the temperature is increased and are simultaneously regenerated when the system is cooled down to room temperature. These ON/OFF molecular valves could be used to overcome multidrug resistance (MDR) in cancer cells and as building blocks for artificial cells.
2:30 PM - BM09.02.04
Incorporating Hierarchical Structure within Hydrogel Biomaterials Using Multifunctional Collagen Mimetic Peptides Toward Directing Stem Cell Fate
Eden Ford1,Amber Hilderbrand1,Chen Guo1,April Kloxin1
University of Delaware1Show Abstract
Extracellular matrix (ECM) properties are important regulators of cell function, particularly at early timepoints during bone healing. For example, physical and chemical properties of the ECM regulate cytoskeletal organization, proliferation, and migration of stem cells to the site of bone injuries for commencing repair. Controlling the presentation of such extracellular cues with molecularly engineered materials provides opportunities to direct bone regeneration. We hypothesize that engineering synthetic hydrogels to recapitulate aspects of the early stages of healing in healthy bone will promote stem cell invasion and remodeling processes toward improving bone regeneration of traumatic fractures or critical-sized defects. To test this, we have created well-defined materials to mimic the mechanical properties, biochemical content, and multiscale structure of native tissues, particularly the collagen-rich environment of the clot-like hematoma formed early in the wound healing process.
We have designed multifunctional collagen mimetic peptides (mfCMPs) that are variants of the Proline-Hydroxyproline-Glycine repeat unit of native collagen. Two variants of this peptide were synthesized: one promoting fibrillar assembly through ionic interactions using charged groups (CMP1a) and the other using hydrophobic interactions of aromatic groups on the C- and N-termini to promote end-to-end assembly (CMP2a). Circular dichroism was used to examine triple helical assembly of the peptides and measure associated melting temperatures, where melting temperatures of CMP1a and CMP2a were determined to be 45.0°C and 60.2°C, respectively. Further peptide assembly and fibril formation was investigated with transmission electron microscopy, where fibrils were observed that mimicked aspects of the hierarchical nanostructure of native collagen. For CMP1a, fibrils approximately 35 nm in width and on the order of 1 μm in length were observed, whereas for CMP2a, fibrils approximately 60 nm in width and on the order of 100 nm in length were observed. Toward studying cell response in vitro, these mfCMPs were covalently crosslinked within cell-degradable poly(ethylene glycol) hydrogels, and rheometry was used to characterize the resulting mechanical properties. Hydrogels with storage moduli in the range of 3500-4500 Pa were generated; further, good cell viability was observed within these unique matrices, with approximately 80% viable cells across conditions.
These studies support our hypothesis that incorporation of mfCMPs within a covalent hydrogel network captures aspects of the fibrillar structure of collagen on both the nano- and microscale toward providing a biomimetic matrix that recapitulates key cues found in ‘soft’ collagenous tissues. Ongoing studies of human mesenchymal stem cells within these materials support their relevance for multidimensional cell culture and suggest that the presence of mfCMPs influences cell-matrix interactions and observed cell response.
2:45 PM - BM09.02.05
Tuning Bioinspired Macromolecular Assembly with Cation-π Interactions
Matthew Gebbie1,Jacob Israelachvili2,J. Herbert Waite2
Stanford University1,University of California, Santa Barbara2Show Abstract
Cation-π interactions govern the assembly of many bio-macromolecules, including the adhesion proteins of marine organisms. Increasingly, cation-π interactions are also implicated in pathological processes, like the formation of neurodegenerative protein aggregates. Thus, developing molecular level approaches for engineering cation-π interactions is of both fundamental and technological importance. Although cation-π bonding has been extensively studied for gas phase ion-aromatic pairs, the energetics of cation-π adhesion in biological and biomineral interfaces, where many binding pairs are in close proximity, remains uncharted. In this seminar, I will discuss using molecular force spectroscopy, supplemented by solid-state NMR measurements, to show that the adhesive properties of simple aromatic- and lysine-rich peptides rival those of the adhesion proteins of the marine mussel. Surprisingly, we find that peptides with the aromatic amino acid phenylalanine, a functional group that is conspicuously rare in mussel proteins, exhibit adhesion that significantly exceeds that of analogous mussel-mimetic peptides. More broadly, we find that interfacial confinement fundamentally alters the energetics of cation-π mediated assembly, an insight that is relevant for diverse areas, from influencing bio-controlled crystal formation to engineering novel bioinspired medical adhesives.
3:30 PM - *BM09.02.06
Nanomaterials for Nervous Regeneration
IRCCS Casa Sollievo della Sofferenza1Show Abstract
Peptidic biomaterials have been receiving great interest because of their easiness of scale-up production, absence of pathogen-transfer risk, biomimetic properties, nanostructured morphology and customization potential for the specific tissue engineering application. However, their proper usage requires the understanding of the multiple-phenomena taking place at different scale levels during self-assembling. In this presentation, focused on the nanotech advancements in the field of nervous regeneration, we will see some multi-disciplinary researches and advances toward the regeneration of spinal cord injuries. This will bring us from coarse-grained molecular dynamics to electro-spinning of self-assembling peptides (SAPs), from cross-linking of SAPs to 3D high-denisty neural stem cells cultures. Lastly, in vivo tests of SAP prosthese in animal models of sub-acute and cronic SCI will be discussed.
4:00 PM - BM09.02.07
Learning from Nature to Form New Organic Materials for Tissue Regeneration
Tel Aviv University1Show Abstract
Molecular self-assembly is a key direction in current nanotechnology based material science fields. In this approach, the physical properties of the formed assemblies are directed by the inherent characteristics of the specific building blocks used. Molecular co-assembly at varied stoichiometry substantially increases the structural and functional diversity of the formed assemblies, thus allowing tuning of both their architecture as well as their physical properties.
In particular, building blocks of short peptides and amino acids can form ordered assemblies such as nanotubes, nanospheres and 3D-hydrogels. These assemblies were shown to have unique mechanical, optical, piezoelectric and semiconductive properties. Yet, the control over the physical properties of the structure has remained challenging. For example, controlling nanotube length in solution is difficult, due to the inherent sequential self-assembly mechanism. Another example is the control of 3D-hydrogel scaffold’s physical properties, including mechanical strength, degradation profile and injectability, which are important for tissue engineering applications.
Here, in line with polymer chemistry paradigms, we applied a supramolecular polymer co-assembly methodology to modulate the physical properties of peptide nanotubes and hydrogel scaffolds. Utilizing this approach with peptide nanotubes, we achieved narrow nanotube length distribution by adjusting the molecular ratio between the two building blocks; the diphenylalanine assembly unit and its end-capped analogue. In addition, applying a co-assembly approach on hydrogel forming peptides resulted in a synergistic modulation of the mechanical properties, forming extraordinary rigid hydrogels. Furthermore, we designed organic-inorganic scaffold for bone tissue regeneration.
This work provides a conceptual framework for the utilization of co-assembly strategies to push the limits of nanostructures physical properties obtained through self-assembly.
Adler-Abramovich, L. et al. Controlling the Physical Dimensions of Peptide Nanotubes by Supramolecular Polymer Coassembly. ACS Nano 10, 7436-7442, (2016).
Halperin-Sternfeld, M., Ghosh, M., Sevostianov, R., Grogoriants, I. & Adler-Abramovich, L. Molecular Co-Assembly as a Strategy for Synergistic Improvement of the Mechanical Properties of Hydrogels. Chem. Comm. 53, 9586-9589, (2017).
Ghosh, M., Halperin-Sternfeld, M., Grigoriants, I., Lee, J., Nam, K. T. & Adler-Abramovich, L. Arginine-Presenting Peptide Hydrogels Decorated with Hydroxyapatite as Biomimetic Scaffolds for Bone Regeneration. Biomacromolecules, 18, 3541–3550, (2017).
Adler-Abramovich, L. et al. Bioinspired Flexible and Tough Layered Peptide Crystals. Adv. Mater. 30, 1704551, (2018).
4:15 PM - BM09.02.08
An Investigation on the Assembly of Particles in a Structurally Colored Protease-Responsive Particle Hydrogel—The Role of Particle Size and Charge
Leopoldo Torres1,John Daristotle1,Omar Ayyub1,Bianca Meinhardt1,Havisha Garimella1,Soenke Seifert2,Nicholas Bedford3,Taylor Woehl1,Peter Kofinas1
University of Maryland College Park1,Argonne National Laboratory2,University of New South Wales3Show Abstract
Pathogens can thrive in an abundance of environments, and pose a significant threat to human health when irrigation or drinking sources become contaminated. The ability to detect the presence of pathogens or biomarkers, such as proteases, using a biosensing platform that is passive and requires no power can help monitor and prevent outbreaks of infectious diseases. We have developed a tunable protease-responsive platform that demonstrated a red-to-blue color shift for all target molecule concentrations between 20 nM and 4000 nM. Structurally colored particle hydrogels were fabricated by centrifuging monodisperse silica particles along with a 4-arm polyethylene glycol (PEG) and a protease-specific peptide linker into a close-packed microstructure, followed by UV irradiation to polymerize the composite. These films swelled in aqueous solutions, and color shift towards the red region of reflected visible light in response to the degree of swelling. Upon degradation of the peptide crosslink, the particles reassembled into a close-packed structure with interparticle spacing less than the initially centrifuged material. This reduction in particle spacing produced a 240 nm color change from the swollen state to the reassembled state of the material for 205 nm particle composites.
To elucidate the mechanism responsible for the color change, we investigated the role of particle size and charge, and polymer concentration in reassembly after degradation. Both particle size and surface functionalization were varied to produce composites with a range of observable structural colors. The reassembled materials reflected shorter wavelengths than their initially fabricated counterparts, indicating that the interparticle spacing had decreased as much as 45 nm for hydrogels with 230 nm particles. In addition, the reassembled composites reflected nearly identical wavelengths independent of the starting polymer weight fraction in the hydrogel. Ultra-small angle x-ray scattering confirmed that the interparticle spacing decreased and the spacing was the same for the reassembled composites. While the particle size or polymer content did not inhibit the reassembly process, particle surface charge was crucial to the reassembly mechanism. Only highly negative (-60mV) particles reassembled to produce structurally colored composites. PEGylated particle hydrogels did not reassemble, and the corresponding composites degraded into the protease solution. Composites with positively charged (+30mV) particle surfaces aggregated irreversibly into a material that appeared white due to incoherent scattering of visible light. Interaction potential models demonstrated that depletion forces provide necessary attraction for reassembly, with a range of up to 120 nm. These findings offer insight into the parameters that will enable passive monitoring of proteases with precise control of structurally colored particle hydrogel responses.
4:30 PM - BM09.02.09
Anti-Biofilm Activity of Graphene Quantum Dots via Self-Assembly with Bacterial Amyloid Proteins
Yichun Wang1,Usha Kadiyala1,Zhi-bei Qu1,Paolo Elvati1,Angela Violi1,Scott VanEpps1,Nicholas Kotov1
University of Michigan1Show Abstract
Bacterial communities, known as biofilms, cause multiple technological and health problems and represent an essential part of Earth’s ecosystem. The environmental resilience and sophisticated organization and of biofilms acting as a multicellular organism is enabled by extracellular matrix (ECM) that creates a protective network of biomolecules around the bacterial communities. The current antibiofilm agents can interfere with ECM production but, being based on small molecules, they can be degraded by bacteria and diffuse away from biofilms, which reduce their efficacy. Here we show that graphene quantum dots (GQDs) can effectively suppress the growth of Staphylococcus aureus biofilms by preventing the self-assembly of amyloid fibers - the essential component of ECM. Mimicking peptide-binding biomolecules, GQDs form supramolecular complexes with phenol soluble modulins (PSMs), the peptide monomers of amyloid fibers. Experimental and computational results show that GQDs dock at the N-terminal of the peptide and change the secondary structure of PSM, which disrupts their fibrillation. Concomitantly, the resulting free PSM monomers turn on biofilm dispersion signaling pathways that enhance the inhibitory effect. The two-prong anti-biofilm activity of GQDs offer a new strategy for manipulation of ECMs of bacterial communities.
4:45 PM - BM09.02.10
Soft to Hard Biomimetic Constructs Using Recombinant Proteins Undergoing Conformational Transition
Hortense Le Ferrand1,Bartosz Gabryelczyk1,Cai Hao1,Ali Miserez1
Nanyang Technological University1Show Abstract
Synthetic mechanical gradients based on synthetic and biocompatible hydrogels currently do not achieve the steep soft to hard transition found in many biological materials like squid beaks or osteochondral cartilage . Indeed, it is difficult to obtain tight molecular packing and high crosslinking density using conventional polymeric building blocks. Here, we employ the recombinantly expressed protein HBP-1 found in the beak of squids, and make use of its folding in presence of polyelectrolytes to expel water and attain high packing density [2,3]. Under acidic pH and in the presence of chitosan, HBP-1 undergoes a conformational transition from predominantly random coil into ß-sheet-rich. At increased ionic strength, this conformation change leads to a phase separation from soluble to liquid droplets and a hydrogel-like phase. At a constant volume fraction of chitosan, the elastic modulus of the HBP-1/chitosan composite increases with the protein content. After drying and cross-linking using catechol chemistry, the resulting organic material shows similar trend under fully hydrated conditions. This observation is reminiscent to what is observed in the native squid beak. Furthermore, concentration gradients can be modeled based on molecular diffusion and phase separation. With this knowledge, gradients of controlled steepness can be obtained. The crosslinked gradient results in an increase of elastic modulus from 0.08 up to 1 GPa despite containing 60 vol% of water. The approach explored here may open new avenues for the fabrication of graded materials based solely on organic biomaterials with potential applications for orthopaedic devices and soft-to-hard attachment in hydrated environments.
 A. Miserez, T. Schneberk, C. Sun, F.W. Zok, J. H. Waite, The transition from stiff to compliant materials in squid beaks, Science, 319, 1816 (2008).
 Y. Tan, S. Hoon, P.A. Guerette, W.Wei, A. Ghadban, C. Hao, A. Miserez, J.H. Waite, Infiltration of chitin by protein coacervates defines the squid beak mechanical gradient, Nature Chemical Biology, 11, 488 (2015).
 H. Cai, B. Gabryelczyk, M.S.S. Manimekelai, G. Gruber, S. Salentinig, A. Miserez, Self-coacervation of modular squid beak proteins – a comparative study, Soft Matter, 13, 7740 (2017).
Chun-Long Chen, Pacific Northwest National Laboratory
Nico Sommerdijk, Eindhoven University of Technology
Tiffany Walsh, Deakin University
Shuguang Zhang, Massachusetts Institute of Technology
Pacific Northwest National Laboratory
BM09.03: Protein-Based Materials
Tuesday AM, November 27, 2018
Sheraton, 2nd Floor, Back Bay A
8:00 AM - *BM09.03.01
CryoTEM Reveals the Molecular Mechanism of Polymorph Selection in Protein Crystallization
Nico Sommerdijk3,Mike Sleutel1,Alexander Van Driessche2
Vrije Universiteit Brussel1, Univ. Grenoble Alpes, CNRS, ISTerre2, Institute for Complex Molecular Systems, Eindhoven University of Technology3Show Abstract
Macromolecular condensed phases such as protein crystals and gels bear great medical, scientific and industrial relevance, yet a molecular understanding of their initial stages of formation is still missing. Insights on the mechanism of nucleation have the potential to resolve one of the longest-standing questions of crystallization, i.e. polymorph selection. To gain control over the emerging polymorph one needs to have a molecular-level understanding of the pathways leading to the various macroscopic states and the underlying selection mechanisms that govern the process. Here we address the issue by capturing protein crystals at birth using time-resolved cryo-transmission electron microscopy and uncover at molecular resolution the nucleation pathways of the protein glucose isomerase into two crystalline and one gelled state. We show that polymorph selection takes place at the earliest stages of transformation and is based on the specific building blocks (monomers and nanorods) for each space group. Moreover, we demonstrate control over the system by selectively forming desired polymorphs through tuning of the directionality and specificity of inter-molecular bonding. These new insights on the mechanisms of nucleation and polymorph selection open new avenues towards the control of macromolecular phase transitions, which is crucial in the further development of protein-based drug delivery systems and macromolecular crystallography.
8:30 AM - BM09.03.02
Building Hierarchically-Ordered 3D Nanomaterials Using 2D Self-Assembling Protein Arrays
Caroline Ajo-Franklin1,Francesca Manea1
Lawrence Berkeley National Laboratory1Show Abstract
Leveraging self-assembly to pattern proteinaceous crystalline arrays in 2D and 3D offers a highly scalable, bottom up approach to develop nanomaterials with new catalytic, optical electronic or structural applications. However, introducing multiple sites of embellishment into existing 2D protein arrays currently utilizes weak interactions that are either sensitive to external conditions or challenging to re-engineer, limiting the ability to program in bifunctionality and new 3D configurations. Here we address these challenges by developing a means to introduce two orthogonal covalent linkages at multiple sites in a highly robust, thermostable 2D crystalline-forming protein. We first engineered the surface-layer (S-layer) protein SbsB from Geobacillus stearothermophilus to display SpyTag or SnoopTag at the C-terminal and two newly-identified locations within SbsB monomer. These regions were able to accommodate SpyTag or SnoopTag peptide tags without affecting the 2D lattice structure. The introduction of tags at distinct locations enabled orthogonal and covalent binding with high precision of SpyCatcher- or SnoopCatcher-protein fusions to micron-sized 2D sheets. By introducing different types of bifunctional crosslinkers, the dual functionalized nanosheets could be programmed to self-assemble into different 3D lamellae, all of which retain their nanoscale order. Additionally, these nanosheets can be functionalized to display two distinct nanomaterial, yielding nanomaterials with emergent optoelectronic properties. Thus, our work creates a modular protein platform that can be facilely programmed to create dual-functionalized 2D and lamellar 3D nanomaterials with novel catalytic, optoelectronic and mechanical properties.
8:45 AM - BM09.03.03
Intracellular Phase-Separated Assemblies of Engineered Disordered Proteins
Ming-Tzo Wei1,Cliff Brangwynne1
Princeton University1Show Abstract
There is currently a growing interest in biopolymer phase transitions, particularly those involving intrinsically disordered proteins/regions (IDPs/IDRs). It has been found that intracellular liquid-liquid phase separations underlie the assembly of many non-membranous organelles such as P granules, nucleoli, and stress granules. However, little is known about the physics of these organelles, including their internal molecular organization and feedback between their molecular and mesoscale properties. Progress on these questions has been hampered by the lack of detailed phase diagrams, which would elucidate how molecular interactions give rise to emergent droplet properties, particularly condensed-protein concentrations and their physical characteristics.
To answer these questions, we investigate the inter-molecular interaction strengths and the full binodal of a phase-separating disordered protein that induces in-vivo phase transitions, utilizing a novel technique, ultrafast-scanning fluorescence correlation spectroscopy. These measurements led to the recent discovery that phase-separated protein droplets have unusually low densities with large void volumes. The data demonstrate how sequence-encoded conformational fluctuations of IDRs give rise to low overlap volume fractions for driving phase separations. Using inter-molecular interactions of native non-membranous organelles, we develop an optogenetic platform that permits light activation of IDR-mediated phase transitions in living cells. Inter-molecular interaction strengths are quantified and demonstrated how IDR sequences determine intracellular phase separation. These studies can elucidate not only physiological phase transitions but also their links to pathological aggregates.
Our results provide a holistic picture of the dynamics and internal organization of phase separated organelles. By uncovering the relationship between molecular level interactions and emergent mesoscale material properties, this work is foundational for understanding the form, function and potential dysfunction of intracellular phase separated assemblies. Our study has significant impact for an extensive community of researchers, with interests spanning biomaterials, bio-inspired materials, macromolecular assembly, self-assembly, intracellular phase separation, disordered proteins dynamics, polymer chemistry, and bioengineering applications of synthesized intracellular biomimetic materials.
9:00 AM - BM09.03.04
Altered Energy-Landscape and Self-Assembly of Protein Crystalline 2D Array at Solid-Liquid Interface
Shuai Zhang1,Robert Alberstein2,F. Akif Tezcan2,James De Yoreo1
Pacific Northwest National Laboratory1,University of California, San Diego2Show Abstract
Protein 2D materials possess diverse sophisticated and synergistic structures, and inherent chemical and biological functions. Harnessing this paradigm of protein 2D materials for bottom-up biomaterial design, synthesis and application is an attractive task with promising perspectives in biomimetic and material science. Inspired by nature cases, various strategies have been developed to construct protein 2D crystals in bulk solution. Recently, computational protein design methodology has considerably improved the structural and functional complexities of protein 2D/3D supramolecular structures from scratch.[1, 2] Besides growth in solution, solid-liquid interface has also been used to template few-layer protein 2D materials. However, the solid-liquid interface that is used to artificially grow protein 2D supramolecular structures is generally limited to supported liquid bilayer. It is still not quite clear how solvent mediated protein-surface interactions to define protein thermodynamics, structure and function at solid-liquid interface. That is the obstacle for artificial design and functional applications of protein 2D crystalline arrays in future.
To address those issues, we assembled the variant of L-rhamnulose-1-phosphate aldolase (RhuA), C98RhuA, with incorporated Cys mutants, into crystalline 2D arrays on solid-liquid interface of mica. By carefully selecting cations and controlling their concentration, we create isotropic protein mono-/bi-layer 2D crystals with controlled packing patterns. It is surprising that the crystallizations of the first and second layers is bimodal that follows non-classical and classical pathways, respectively. We also proved that solvent mediated protein-surface interactions can alternate the energy-landscape of protein self-assembly from that in bulk to stabilize the original intermediate and quasi stable phase. C98RhuA can epitaxially grow on top of the surface that has different symmetry. All the findings inspire the novel strategy to synthesize protein crystalline 2D arrays at solid-liquid interface artificially. They also help to elucidate the growth modal of protein 2D architectures at solid-liquid interface. They remind us the importance of solvent mediated surface templating in the self-assembly of protein 2D structures both in nature and in human manner.
1. Huang, P.-S., S.E. Boyken, and D. Baker, The coming of age of de novo protein design. Nature, 2016. 537: p. 320.
2. Gonen, S., et al., Design of ordered two-dimensional arrays mediated by noncovalent protein-protein interfaces. Science, 2015. 348(6241): p. 1365.
3. Suzuki, Y., et al., Self-assembly of coherently dynamic, auxetic, two-dimensional protein crystals. Nature, 2016. 533(7603): p. 369-373.
9:15 AM - BM09.03.05
Microbial Factories for Programmed Production of Functional Biomaterials
Avinash Manjula Basavanna1,2,Anna Duraj-Thatte1,2,Neel Joshi1,2
Wyss Institute for Biologically Inspired Engineering1,Harvard University2Show Abstract
Biological systems are highly complex and sophisticated with unparalleled structure-function correlations. Remarkably, biological systems produce materials with extraordinary properties and functions under ambient conditions, which is in total contrast to humans’ heat-beat-treat strategies. Thus, the capabilities of a technology by which biological networks of a cell can be programmed, offers tremendous potential as cellular factories and to produce biomaterials for various functional applications.
In this regard, we employ a novel technology entitled Biofilm-Integrated Nanofiber Display (BIND) that focuses on the curli system-the primary proteinaceous structural component of E. coli biofilms. Curli are highly robust functional amyloid nanofibers (diameter 4-7 nm) formed by the extracellular self-assembly of a small (13 kDa) secreted protein, CsgA. By genetic engineering, artificial peptide domains were grafted to the amyloid protein CsgA and the resulting CsgA fusion proteins were successfully secreted from the E. coli cells. Remarkably, these engineered fusion proteins were found to extracellularly self-assemble into amyloid nanofiber networks and also exhibited the characteristic functions of the grafted artificial peptide domains. By using BIND technology, E. coli biofilm matrix is conferred with several artificial functions for nanomedicinal, nanomechanical and nanoelectronics applications.
10:00 AM - *BM09.03.06
S-Layers—Principles and Applications
Uwe Sleytr1,Dietmar Pum1
Univ Bodenkultur1Show Abstract
One of the key challenges in nanobiotechnology is the utilization of self-assembly systems wherein molecules spontaneously associate into reproducible aggregates and supromolecular structures. In this contribution, the basic principles of crystalline bacterial surface layers (S-layers) and their use as patterning elements will be described. The broad application potential of S-layers in nanobiotechnology is based on the specific intrinsic features of these monomolecular arrays which are composed of identical protein or glycoprotein subunits. Most important, physicochemical properties and functional groups on the protein lattice are arranged in well-defined positions. Many applications of S-layers depend on the capability of the isolated subunits to recrystallize into monomolecular arrays in suspension or on suitable surfaces (e.g. polymers, metals, silicon wafers) or interfaces (e.g. lipid films, liposomes, emulsomes). S-layers also represent a unique structural basis and patterning element for generating more complex supramolecular structures involving all major classes of biological molecules Thus, S-layers fulfil key requirements as building blocks for the production of new supramolecular materials and nanoscale devices as required in nanobiotechnology and synthetic biology.
Sleytr, U.B., Schuster, B., Egelseer, E.M., Pum, D. (2014) FEMS Microbiol Rev, 38, 823-864.
Pum, D., Toca-Herrera, J.L., Sleytr, U.B. (2014) Nanotechnology, 25, 312001.
Sleytr, U.B. 2016. Curiosity and Passion for Science and Art. World Scientific. ISBN 9813141816
We acknowledge the financial support by the Air Force Office of Scientific Research (AFOSR) (Grant FA9550-15-1-0459).
10:30 AM - BM09.03.07
Controlled Formation of Enzyme-Scaffold Complex for Biocatalysis Using a Self-Assembling Protein Template
Samuel Lim1,Florence Barraud1,Sophia Prem1,Dominic Glover2,Douglas Clark1
University of California, Berkeley1,University of New South Wales2Show Abstract
In nature, enzymes that catalyze multi-step reactions are often organized in close proximity to allow the efficient channeling of intermediates from one active site to another. Diverse synthetic scaffolds based on DNA and proteins have been designed to mimic such spatial control of enzymes, and have proven successful in facilitating cascade reactions. Recent studies revealed that such enhanced catalysis can also result from formation of enzyme agglomerates rather than direct channeling of intermediates between adjacent enzymes, highlighting the need to engineer the interactions that define metabolic clusters. Thus, there is a demand for scaffolds that can effectively crosslink with each other to form higher-order structures in a programmable manner, in addition to simply templating the enzymes.
The g-prefoldin (gPFD) is a filamentous protein isolated from the hyperthermophilic archaeon Methanocaldococcus jannaschii; its remarkable stability, unique modularity, and self-assembly into filaments with chaperone activity render it an ideal building block for the bottom-up construction of functionalized protein nanostructures. Here we propose a strategy to utilize the gPFD to build enzyme agglomerates in tunable fashion. Using the combinations of orthogonal protein-peptide bioconjugation pairs, the gPFD filaments displaying the peptide tags are first conjugated with the enzymes, and then crosslinked using the linker proteins. Thus, the extent of crosslinking is tunable through simply varying the stoichiometry between the scaffold and linker proteins.
We verified the gPFD scaffold’s ability to cluster enzymes in proximity using FRET analysis of the filaments containing fluorescent protein pairs. Subsequently, we investigated the effect of agglomerate formation on the catalytic activities of multi-step reactions using two different model system pairings: glucose oxidase (GOX)-horseradish peroxidase (HRP) and alcohol dehydrogenase (ADH)-aldehyde dehydrogenase (ALDH). Ultimately, the ability to fabricate enzyme-scaffold complexes with programmable stoichiometry and dimensions will enable better control over single- and multi-step enzymatic catalysis.
11:00 AM - BM09.03.09
Mimicking Dividing Cells by Assembly of Protein Structures Inside Aqueous Two-Phase Droplets
Anderson Shum1,Yang Song1,Tuomas Knowles2,Thomas Michaels2
University of Hong Kong1,University of Cambridge2Show Abstract
In this work, we demonstrate that assembly of macromolecules, such as proteins, can cause aqueous droplets to exhibit division, even in the absence of a cell membrane. The all-aqueous nature of the systems results in tunable interfacial tension, affinity partitioning and osmotic responses. The solubility of different types of macromolecules across the interfaces enables new strategies to assemble structures at the droplet interfaces. While the significantly lower interfacial tension can make stabilization of the interface difficult due to the slow adsorption dynamics by surfactants and particles, structures that have been assembled at the interfaces can be easily expelled. This contributes to the more sophisticated dynamics of the hierarhically structured all-aqueous droplets. These droplets have great potential to be utilized as templates for fabricating materials with novel properties.
11:15 AM - BM09.03.10
Self-Assembly of Elastin-b-Collagen-Like Conjugates Mediated by Triple Helical Parameters
Lucas Dunshee1,Kristi Kiick1,Millicent Sullivan1
University of Delaware1Show Abstract
Physiochemical irregularities within extracellular matrix (ECM) proteins such as collagen can lead to a wide range of connective tissue disorders including osteogenesis imperfecta and osteoarthritis. Current pharmaceutical regimens to treat such diseases suffer from off-target effects, suggesting that new approaches for targeted delivery are necessary. In the last decade, ECM-inspired polypeptide materials have garnered significant interest for their ability to selectively mimic specific matrix components such as collagens and elastins, offering new opportunities to control drug delivery within specific tissues. For example, triple helix forming collagen-like peptides (CLPs) comprising (Gly-Pro-Hyp)n amino acid repeats can hybridize with high efficiency to denatured collagen proteins in the body via thermal annealing of peptide and protein single strands into a stable triple helix. Additionally, elastin-like peptides (ELPs) that consist of (Val-Pro-Gly-XAA-Gly)n (where XAA is any amino acid with the exception of proline) amino acid repeats possess a lower critical solution temperature in which aggregation occurs upon heating above this temperature, making ELPs ideal candidates for on demand drug delivery behavior. Recently, our group has reported on the design of hybrid peptides with linked CLPs and ELPs, and the assembly of thermoresponsive, elastin-b-collagen-like peptide nanovesicles that are capable of dissociating at high temperature (70°C). These nanovesicles offer intriguing potential in drug delivery applications due to their dual thermoresponsivity and inherent ability to bind to degraded collagen protein. However, in order to make an ELP-CLP nanoparticle with optimal drug delivery properties such as physiologically relevant hybridization to degraded collagen protein, the critical parameters of their self-assembly must first be understood, specifically with respect to the CLP domain. To test the effects of the triple helical (CLP) melting temperature on temperature-dependent nanovesicle assembly and dissociation behavior a small library of ELP-CLP conjugates was made with varying numbers of CLP (G-X-Y) repeats and varied CLP sequences. These conjugates were characterized for their thermoresponsivity and their ability to form self-assembled structures. The melting temperature, repeat length, and overall hydrophilicity of the CLP domain were found to be of critical importance to nanoparticle formation.
11:30 AM - *BM09.03.11
Soft Functionalization of Silk Fibroin Materials and Bio-Flexible Devices
Xiang Yang Liu1,2
National University of Singapore1,Xiamen University2Show Abstract
As an excellent flexible biomaterial, Bombyx mori silk fibroin materials offer exquisite mechanical, optical, and electrical properties which are advantageous toward the development of next-generation biocompatible electronic devices. In this concern, to re-engineer the hierarchical structure of soft materials and to functionalize the materials are the two common approaches to achieve the functions. This requires the synergy of structures among different levels, which include the re-construction of the hierarchical structure of soft/SF materials at the mesoscale and or Mesoscopic Material Assembly (MMA), which is to add and bind some specific nanomaterials or molecule to the networks so as to acquire some additional functions without jeopardizing the original performance. In this talk, I will cover the principles and strategies of mesoscopic structural re-engineering and functionalization of SF materials, which allows in the design and integration of high-performance bio-integrated devices for future applications in consumer, biomedical diagnosis, and human–machine interfaces.
BM09.04: Bio-Inspired Materials Based on DNA or Peptide Building Blocks
Xiang Yang Liu
Tuesday PM, November 27, 2018
Sheraton, 2nd Floor, Back Bay A
1:30 PM - *BM09.04.01
Colloidal Crystal Engineering with DNA—Creating a Genetic Code for Materials Design
Northwestern University1Show Abstract
The materials-by-design approach to the development of functional materials requires new synthetic strategies that allow for material composition and structure to be independently controlled and tuned on demand. Although it is exceedingly difficult to control the complex interactions between atomic and molecular species in such a manner, interactions between nanoscale components can be encoded, independent of the nanoparticle structure and composition, through the ligands attached to their surface. DNA represents a powerful, programmable tool for bottom-up material design. The Mirkin Group has shown that DNA and other nucleic acids can be used as highly programmable surface ligands (“bonds”) to control the spacing and symmetry of nanoparticle building blocks (“atoms”) in structurally sophisticated materials, analogous to a nanoscale genetic code for material assembly. The sequence and length tunability of nucleic acid bonds has allowed us to define a powerful set of design rules for the construction of nanoparticle superlattices with more than 30 unique lattice symmetries, spanning over one order of magnitude of interparticle distances, with several well-defined crystal habits. Further, this control has enabled exploration of sophisticated symmetry breaking processes, including the body-centered tetragonal lattice as well as the clathrate lattice, the most structurally complex nanoparticle-based material to date (>20 particles per unit cell). The nucleic acid bond can also be programmed to respond to external biomolecular and chemical stimuli, allowing structure and properties to be dynamically tailored. Notably, this unique genetic approach to materials design affords functional nanoparticle architectures that can be used to catalyze chemical reactions, manipulate light-matter interactions, and improve our fundamental understanding of crystallization processes.
2:00 PM - BM09.04.02
DNA-Programmed Assembly of Single Crystalline Nanoparticle Superlattices at Interfaces
Massachusetts Institute of Technology1Show Abstract
The programmability of DNA makes it an attractive structure-directing ligand for the assembly of nanoparticle superlattices with unique structure-dependent physical phenomena. While DNA base pairing has enabled the development of materials with nanometer-scale precision in nanoparticle placement and independent control over particle size, lattice parameters, and crystal symmetry, manipulating the macroscopic shape of the lattices remains challenging. By pairing this “bottom-up” assembly method with “top-down” lithographic techniques and assembling nanoparticle superlattices on a patterned substrate, complete control over crystal size, shape, orientation and unit cell structure can be realized. The key challenges in developing this technique are to first understand how different design factors affect the assembly process in this broken-symmetry system that is assembled at an interface, and subsequently develop structure-property relationships that correlate the above mentioned design parameters with the resulting overall material structure. Here, we examine both at-equilibrium deposition processes capable of generating single crystals with well-defined shapes, as well as post-deposition annealing to transform disordered particle arrangements into crystalline arrays. Using a combination of X-ray diffraction and electron microscopy techniques, both surface morphology and internal thin film structure are examined to provide an understanding of the mechanisms of particle crystallization under conditions where crystal growth is anisotropic due to a boundary condition. This novel method for controlling particle assembly draws several strong analogies to traditionally atomic epitaxy/heteroepitaxy, providing a useful tool for understanding thin film growth processes. As a result, we are able to realize 3D architectures of arbitrary domain geometry and size, thereby making materials with unprecedented precision across multiple length scales.
2:30 PM - BM09.04.04
Magnesium Stabilized Multifunctional DNA Nanoparticles for Tumor-Targeted and pH-Responsive Chemotherapy
South University of S&T of China1Show Abstract
Functional nucleic acids, that can target cancer cells and realize stimuli-responsive drug-delivery in tumor microenvironment, have been widely applied for anti-cancer chemotherapy. The high cost, unsatisfactory biostability, and complicated fabrication process are the main limits for the development of DNA-based drug-delivery nanocarriers. Recently, a kind of DNA-MgPPi (magnesium pyrophosphate) composite nanoparticles has been produced from rolling circle amplification (RCA), which combine advantages of the designable and high-throughput isothermal amplification technique and the high stability of DNA condensation structures, quickly becoming an attractive biomedical material with great potentials. Herein, instead of using MgPPi, we found that only Mg2+ is sufficiently enough to stabilize the functional DNAs for chemotherapeutic applications. The very long single-stranded RCA product with a high charge density is more prone to form a stable condensation structures compared with a short oligonucleotide. Moreover, the dynamic electrostatic interactions between Mg2+ and DNA can better preserve the functions of DNA, which is more suitable for the design of drug-delivery system. A tumor-targeting Dox-delivery nanoparticle (~ 100 nm) was synthesized by the condensation of RCA products in the presence of an excessive amount of Mg2+, which showed good bio-stability in serum, considerable Dox loading capability, specific cancer-targeting ability, and pH-responsive sustained Dox release. The DNA nanoparticle not only has a simple composition, but also it will keep intact after the excessive exterior Mg2+ is removed, making it safe and ideal for in vivo application. Through cellular and in vivo experiments, we thoroughly demonstrated that this kind of Mg2+ stabilized multi-functional DNA nanoparticles can successfully realize tumor-targeted Dox delivery.
2:45 PM - BM09.04.05
DNA-Programmable Nanoparticle Lattices Assembled on Polymer-Patterned Surfaces
Sha Sun1,2,Dmytro Nykypanchuk2,Gregory Doerk2,Charles Black2,Oleg Gang2,3,Diana Lopez2
Xi'an Jiaotong University1,Brookhaven National Laboratory2,Columbia University3Show Abstract
Photonic and electronic devices require precise control of functional components at the nanosacle. The development of DNA nanotechnology offers a fascinating platform to direct the assembly of nanoparticles into well-organized architectures with prescribed distances and spatial arrangements. Here, we combine DNA-based assembly and diblock copolymer self-assembly to realize the multi-layer assembly of gold nanoparticles into large-area, three-dimensional arrays. Specifically, the assembly of gold nanoparticles is directed through binding with DNA origami that forms arrays, and the array growth is controlled by patterns formed via diblock copolymer on the surface. In our approach, DNA-programmable nanoparticle lattices are sequentially assembled with registry of polymer pattern. We show the potential to assemble functional nanoparticles in layer-by-layer manner with controllable interlayer distance and in-plane arrangements through a combination of surface patterns and DNA nanostructures.
3:30 PM - *BM09.04.06
Engineering Molecular Assembly for 3D Electronics
Thom LaBean1,Nikolay Frick1,Ming Gao1
North Carolina State University1Show Abstract
The ability to design and program complex molecular interactions between synthetic biomolecules (especially polynucleotides and polypeptides) has led to a revolution in artificial nanomaterials capable of self-assembly. For example, DNA-based nanotech entails the design of artificial nucleotide sequences capable of self-assembling into desired geometric shapes and patterns with nanometer-scale precision. These synthetic DNA nanostructures have been shown useful for organizing other materials including inorganic nanoparticles (metals and semiconductors), nucleic acid aptamers, and carbon nanostructures. We are working with DNA self- and directed-assembly to develop a general purpose molecular assembly toolbox useful for a wide variety of applications, especially in nanoelectronics and medicine. One promising future direction is the bottom-up fabrication of electronics components and devices including molecular assembly of wires and metal nanoparticles toward the construction of single-electron transistors, multicomponent devices, and artificial neural networks.
4:00 PM - BM09.04.07
Self-Assembled Peptide Nano-Materials for Optics and Electronic Applications
Sharon Gilead1,Ehud Gazit1
Tel Aviv University, Department of Molecular Microbiology and Biotechnology1Show Abstract
In recent years, a key direction in the field of electronics and electro-optics involves the transition from inorganic to organic components, including organic light emitting diodes (OLED), thus paving the way towards flexible and wearable electronic and light emitting devices. Bio-inspired organic materials may be the next-generation of organic optoelectronic devices based on self-organization principles, which allow facile synthesis, eco-friendliness, resistance to oxidation and no need for heavy metal doping.
Recent advances in bioorganic nanotechnology have established the notion that very simple building blocks, such as dipeptides, can form regular nanostructures with distinct mechanical, optical, piezoelectric and electronic properties. In particular, members of the diphenylalanine (FF) peptide archetypical family have been shown to form various morphologies and ordered nanostructures such as tubes, rods, fibrils, spheres, plates and macroscopic hydrogels with nano-scale order.
Several studies have explored the piezoelectric properties of the diphenylalanine (FF) peptide. In the presence of an external electric field, vertically aligned FF microrod arrays can be organized on a substrate, resulting in enhanced piezoelectric response.
Here we show the ability of FF and other similar peptide assemblies to be used in various electronics and optics application as new bioorganic materials. FF assemblies can act as an active optical waveguiding material, allowing locally excited states to propagate along the axis of the assemblies. In addition, Fmoc capped building blocks exhibit remarkable optical properties, such as quantum confinement and fluorescence. Other rod-like assemblies and toroid-like assemblies exhibit remarkable physicochemical features, including high thermal stability, metallic-like mechanical rigidity, luminescence, piezoelectricity and semi-conductivity.
The ability of FF to self-assemble into ordered structures was discovered by a systematic reductionist exploration of biological recognition modules in an amyloidogenic polypeptide. We are applying a similar reductionist approache to expand our search for minimal building blocks towards single amino acids as well as other metabolites such as nucleobases, demonstrating their self-assembly into various ordered structures. Doing this we are enlarging our library of biological building blocks which bear the potential to be novel bio-inspired supramolecular materials for Optics And Electronic applications.
4:30 PM - BM09.04.09
DNA Origami-Assembled Light-Emitting Nanoclusters with Controllable Optical Output
Honghu Zhang1,Mingxing Li1,Kaiwei Wang1,2,Ye Tian1,Jia-Shiang Chen1,Mingzhao Liu1,Katherine Fountaine3,Donald DiMarzio3,Mircea Cotlet1,Oleg Gang1,4
Brookhaven National Laboratory1,Xi'an Jiaotong University2,Northrop Grumman Aerospace Systems3,Columbia University4Show Abstract
Structural DNA nanotechnology has emerged as a powerful method to fabricate targeted nanoscale architectures. Using rationally designed DNA origami frames, nanoparticles can be coordinated in a prescribed manner in 3D. Here, we have designed DNA origami frames for assembling various nanoparticles in pre-determined locations. The DNA frames have enabled well-defined nanocluster assembly with nanometer-precision positioning, and controllable high-purity stoichiometry with tunable functionality. We have fabricated DNA origami-constructed nanoparticle clusters, consisting of spherical quantum dots (QDs) and gold nanoparticles (AuNPs) that exhibit controllable photoluminescence (PL) when the excitation wavelength is close to surface plasmon resonance of the AuNPs. Furthermore, these self-assembled nanoclusters emit highly polarized light. By varying the size and number of AuNPs in the nanoclusters, we have explored correlations between the assembled structures and the PL polarization magnitude and the overall PL enhancement. Our DNA origami based nanoclusters with precisely built 3D architectures provide an efficient route to control single emitter optical output.
4:45 PM - BM09.04.10
Reconfigurable Nanoparticle Superlattices with Tunable DNA Bonds
Jinghan Zhu1,Youngeun Kim1,Haixin Lin1,Shunzhi Wang1,Chad Mirkin1
Northwestern Univ1Show Abstract
Stimuli-responsive nanomaterials with reconfigurable structures and properties have garnered significant interest in the fields of optics, electronics, magnetics, and therapeutics. DNA is a powerful and versatile building material that provides programmable structural and dynamic properties, and indeed, sequence-dependent changes in DNA have already been exploited in creating switchable DNA-based architectures. However, rather than designing a new DNA input sequence for each intended dynamic change, it would be useful to have one simple, generalized stimulus design that could provide multiple different structural outputs. In pursuit of this goal, we have designed, synthesized, and characterized pH-dependent, switchable nanoparticle superlattices by utilizing i-motif DNA structures as pH-sensitive DNA bonds. When the pH of the solution containing such superlattices is changed, the superlattices reversibly undergo: (i) a lattice expansion or contraction, a consequence of the pH-induced change in DNA length, or (ii) a change in crystal symmetry, a consequence of both pH-induced DNA “bond breaking” and “bond forming” processes. The introduction of i-motifs in DNA colloidal crystal engineering marks a significant step toward being able to dynamically modulate crystalline architectures and propagate local molecular motion into global structural change via exogenous stimuli.
BM09.05: Poster Session I: Bioinspired Materials
Wednesday AM, November 28, 2018
Hynes, Level 1, Hall B
8:00 PM - BM09.05.01
Tandem Molecular Self-Assembly in Liver Cancer Cells
Jie Zhan1,Yanbin Cai1,Ling Wang2,Zhimou Yang1
College of Life Sciences, Nankai University1,College of Pharmacy, Nankai University2Show Abstract
Inspired by nature, stimuli-responsive self-assembly has been widely explored for spatiotemporally regulating diverse cellular functions. In situ formation (both pericellular and intracellular) of assemblies of man-made small molecular in cell milieu has been successfully applied for controlling the cell behavior and fate. The differences of the expression levels of bio-signals (i.e., enzyme or small molecule) between cells are favorable natural-source of inspiration for designing precursors to form sophisticated assemblies with enhancing selectivity to target and inhibit diseased cells. We herein describe the tandem molecular self-assembly of a peptide derivative NBD-GFFpY-ss-ERGD (Tandem Molecular Self-assembly Precursor, TMSP) that is controlled by a combination of enzymatic and chemical reactions. In phosphate-buffered saline (PBS), TMSP self-assembles first into nanoparticles by phosphatase and then into nanofibers by glutathione. Liver cancer cells exhibit higher concentrations of both phosphatase and GSH than normal cells. Therefore, the tandem self-assembly of TMSP also occurs in the liver cancer cell lines HepG2 and QGY7703; TMSP first forms nanoparticles around the cells and then forms nanofibers inside the cells. Owing to this self-assembly mechanism, TMSP exhibits large ratios for cellular uptake and inhibition of cell viability between liver cancer cells and normal liver cells. We envision that using both extracellular and intracellular reactions to trigger tandem molecular self-assembly could lead to the development of supramolecular nanomaterials with improved performance in cancer diagnostics and therapy.
8:00 PM - BM09.05.02
Fabrication and Evaluation of a Repairable Resistive Device Using Bio Material for a Synaptic Device
Takahiko Ban1,Yukiharu Uraoka2,Shin-ichi Yamamoto1
Ryukoku University1,NAIST2Show Abstract
As our information society advances, the roles of semiconductor devices are becoming increasingly important. However, the total volume of data handled by humans is steadily expanding and becoming complex. The volume of data has been estimated to reach 44 zettabytes in 2020. Devices that can record and process large volumes of information are required for the benefit of society. However, in forthcoming practical nanoscale applications, the downsizing of the device reaches its limit. Therefore, it is necessary to develop new devices with different principles and structures. As one solution to expand the scaling limit and to process information more flexibly, devices and/or circuits that simulate a human brain have gained attention. Simulating a human brain in computers is expected to enhance recognition capability and reduce power consumption. Research on devices reproducing synapses, which transmit information, is attracting particular attention. Neural networks in the human brain comprise neurons connected with each other through synapses. A synapse connects neurons upon receiving a stimulus; the weaker the stimulus, the weaker the connection. Resistive memory has been proposed as a circuit capable of simulating a synapse. In this research, resistive switching memories (ReRAM) were fabricated for synaptic devices. By applying repair capacity to the ReRAM, the resistance value is returned to original value over time under low bias. In addition, the ReRAM are fabricated with nanoparticles (NPs) using biomaterials. The biomaterial is a spherical shell protein called ferritin. It has the ability to precipitate inorganic substances as NPs in the internal cavity, and various placement methods can be provided by modifying its surface. In this study, a method is adopted in which NPs are evenly arranged at intervals of 40 to 50 nm by modifying PEG on the surface of ferritin.
A tantalum oxide (Ta2O5) film was deposited on the lower electrode deposited by electron beam deposition (EB depo.), and the upper electrode was similarly deposited to fabricate a usual ReRAM. A structure in which manganese oxide (MnO2) was sandwiched between the Ta2O5 film and the lower electrode was fabricated to prepare a ReRAM having repairing capability. In this device, we succeeded in developing a device that returns to the original high resistance state with low bias voltage and time. It is short-term plasticity well-known as synaptic movement. Also, manganese oxide is nanoparticulated by ferritin. The resistance change phenomenon, which is a complex analog operation, is limited to only nanoparticles. By preparing this device, it is expected to mimic the work of actual synapses which exist in tens of thousands among neurons.
8:00 PM - BM09.05.03
Surface Interactions of DNA and Mononucleotides with Sol-Gel Derived Silica Host
Caner Durucan2,3,Derya Kapusuz1
Gazinantep University1,METU2,BIOMATEN3Show Abstract
Double stranded DNA and dAMP (2′-Deoxyadenosine 5′-monophosphate) were encapsulated in silica by sol-gel route. The microstructure of the biomolecule-hostings gels and the chemical interactions between biomolecules and silica host have been investigated. Ethidium bromide (EtBr) intercalation and leach out tests showed revearled a high hydraulic reactivity for encapsulated DNA and dAMP gels due to presence of more silanol groups than plain silica gel. For both biomolecules, no chemical binding occurred with Si core of the silica network. The chemical association between DNA/dAMP and silica host was through phosphate groups and molecular water attached to silanols, acting as a barrier around biomolecules. The helix morphology was found not to be essential for such interaction. BET analyses showed that interconnected, inkbottle shaped mesoporous silica network with an average pore size of 5.6 nm for DNA and 4.8 nm for dAMP containing bulk gels, respectively.
8:00 PM - BM09.05.05
Fabrication of Composite Coatings for Drug Delivery Using Bile Acid Salts
Amanda Clifford1,Igor Zhitomirsky1
McMaster University1Show Abstract
A conceptually new fabrication technique has been developed for the fabrication of composite gel coatings containing commercially available drugs that exhibit low solubility in water. This technique utilizes bile acid salts, which are natural anionic biosurfactants, that have powerful solubilizing properties; these can be used for the dispersion of organic molecules, dietary fat and vitamins in mammalian digestive systems. This work is of paramount importance, as 40% of commercially available drugs and 90% of drugs being developed exhibit low solubility in water. Bile acid salts have also been found to form unique porous gel structures, which is advantageous for tissue scaffold applications. In the present work, bile acid salts were used for the solubilization, dispersion, charging and composite film formation using anodic electrophoretic deposition (EPD). Ibuprofen, which is a commercially available non-steroidal anti-inflammatory drug, and tetracycline, a clinically used anti-biotic were used as model water-insoluble drugs. Cholic acid sodium salt and deoxycholic acid sodium salt were used as model bile acid salts. In our method, bile acid salts dissolved in an aqueous suspension solubilized our model water-insoluble drugs to form charged mixed micelles. Under the influence of electric field, the mixed micelles electromigrate to the anode surface, where the carboxylic groups of the bile acid salts become protonated and form an insoluble film on the electrode surface. Composite films were obtained by potentiodynamic and galvanostatic deposition methods. The cyclic voltammetry and quartz crystal microbalance data provided an insight into the deposition mechanism and kinetics of deposition at various conditions. X-ray analysis confirmed the formation of composite films. SEM investigations of pure bile acid films and composites revealed the influence of the co-deposited drugs on film microstructure. The composite coatings were deposited as functionally graded materials or laminates for controlled drug delivery. This work offers a versatile approach for the deposition and delivery of drugs, which have poor solubility in water. We demonstrate that new EPD strategies pave the way for the fabrication of composite films containing drugs and other advanced functional biomaterials. These films may be used for a variety of applications: from tissue scaffolds to orthopaedic coatings.
8:00 PM - BM09.05.07
Studies on Synthesis and Fluorescence Spectroscopy of Hybrid Magnetic Nanoparticles (Fe3O4-Au) Linked with Fluorescent Molecule
Alisha Memon1,Andrew Nunez1,Mostafa Sadoqi1,Elmustapha Feddi2,Gen Long1
Saint John's University1,ENSET, Mohammed V University2Show Abstract
Magnetic and fluorescent nanoparticles are widely studied in biomedical research for their use in drug delivery, cell separation, magnetic resonance imaging (MRI), hyperthermia, various multimodal techniques, etc. In this study, we report a recent work on nanoparticles incorporating with a fluorescent molecue and a superparamagnetic core via nanoscale engineering. Fe3O4-Au hybrid nanoparticles are synthesized via a solution phase chemical reaction in an inert N2 atmosphere. These synthesized hybrid nanoparticles are characterized by UV-Vis-NIR spectroscopy, fluorescence spectroscopy, XRD, TEM, etc. Optimal synthesis conditions are also highly relevant in producing stable and uniform hybrid nanoparticles without impurities. Fluorescence spectroscopy show the correlations between the lifetime and intensity of fluorescence and sizes, compositions, shapes of hybrid nanoparticles as well as the conjugation process to link the nanoparticles to fluorescent molecules. By carefully engineering the growth conditions, such as altering growth temperatures and precursor reagent ratios, functional hybrid magnetic nanoparticles can be optimized for hyperthermia, MRI and other multimodal biomedical applications.
8:00 PM - BM09.05.09
Mineral-Assisted Self-Assembled Nanostructures from Poly-Glycine, Hydrogels of Short Peptides and Alpha-Hydroxy Acids
Rehana Afrin1,Tony Jia1,James Cleaves1,Taka-aki Yano2,Masahiko Hara2,1
Earth-Life Science Institute (ELSI), Tokyo Institute of Technology1,School of Materials and Chemical Technology, Tokyo Institute of Technology2Show Abstract
The principle of self-assembly is fundamental in the formation of higher order structures from small molecules. Many kinds of such structures have been formed from small amino acids and short peptides as useful materials for bio-medical purposes . They also have a fundamental importance as the starting ingredients for the creation of life on the Earth. From this point of view, we first studied the adsorption mechanism of amino acids and peptides to solid surfaces  and proceeded to investigate the formation of self-assembled structures in solution and on solid surfaces. In this study, we present the formation of new types of self-assembled structures of poly-glycine, short peptides and alpha-hydroxyacids and show their interesting structural properties obtained with the atomic force microscope (AFM).
Poly-glycine is water insoluble but soluble in tetrafluoroacetic acid (TFA). Dilution of its TFA solution with deionized water led to the formation of small self-assembled structures. AFM observation revealed the formation of thin and flat films (1 – 2 nm thick and 200 – 500 nm wide) and isolated fibers (10 – 50 nm wide). Because poly-Gly does not have charges except for at its N- and C-termini, some types of rather strong non-ionic attractive force, most likely involving van der Waals force, must be working at the basic level. The thickness of the film implies an alignment of a few poly-Glycine helices in the vertical direction and many of them into an ordered side-side arrangement. Such a uniformly flat structure suggests its possible role as a reliable and well defined platform for further assemblage with other molecules as a composite film.
We also found that some short peptides and alpha-hydroxy acids self-assembled into long strings as well as a hydrogel structure under an aqueous condition at specific pHs. The hydrogel has a nano-mesh like structure that can be reconstructed on mineral surfaces and visualized with AFM. We are particularly interested in possible roles of these structures as potential functional biomaterial in the origin of life on the Earth.
8:00 PM - BM09.05.10
Catalytic Self-Assembly of Peptidic Bolaamphiphiles Coordinated with Transition Metal Cofactors
Sang-Yup Lee1,Min-Chul Kim1,Changjoon Keum1
Yonsei Univ1Show Abstract
Peptidic bolaamphiphile is an biomimetic amphiphilic molecule whose biochemical activity can be tuned by the designer peptides. The peptidic bolaamphipihles have peptide or amino acid segments as hydrophlic moieties that are associated with the central alkyl chain to display amphiphilic property. Similar to other amphiphilic molecules, these peptidic bolaamphiphiles self-assemble to form complex structures while exposing the biological segment to the surface in an aqueous medium. Here, assembled structure of histidyl bolaamphiphiles was exploited as a biomimetic host matrix whose histidine imidazoles are exploited as ligands to coordinate with transition metal ions. By coordinating with transition metal ions, metalloenzyme-mimetic catalysts could be built. In particular, catalytic activity for CO2 hydration and oxidation of organic compounds could be realized by coordinating various transition metal ions to the histidyl bolaamphiphile assembly. Furthermore, the catalytic water evolution was achieved by introducing Iridium to the histidyl bolaamphiphile assembly. The prepared metal-bolaamphiphile catalyst was surveyed with spectroscopic studies to verify the origin of the catalytic activity. This assembly of peptidic bolaamphiphiles will be beneficial for the building of catalysts mimicking various metalloenzymes.
8:00 PM - BM09.05.11
Thermodynamic Properties of Pluronic F127 Micelles with Added Cefepime Determined by Differential Scanning Calorimetry
Lydia Mensah1,Brian Love1
University of Michigan–Ann Arbor1Show Abstract
Aqueous amphiphilic copolymer polyether solutions have been made with varying amounts of a third constituent with the notion of forming drug loaded gels. Our research group is interested in how additive molecules perturb the structure of amphiphilic copolymers that are known to form colloidal crystals. We are investigating whether likely correlations exist between how strongly the drug interacts within the hydrophobic and hydrophilic regions of micelles and colloidal crystals and how bioavailable the drug is as it elutes from within the gel. We have made 25% aqueous solutions of PEO-PPO-PEO copolymers (F-127, BASF) formulated and tested between -5oC and 50oC. Rheology, and DSC have been the primary tools of measurement and we have focused our initial efforts on cefepime to observe how its presence affects micelle formation and colloidal crystallization. Drug-loaded polymers are of interest as schemes within the drug delivery community for controlled release and other studies have been done using dendrimers, and responsive polymers contained within other polymers. We have found that when Cefepime has been added in concentrations ranging from 2-8% of the mass of a 25% PEO-PPO-PEO copolymer solution, the onset temperature for micelle formation systematically drops with increasing cefepime concentration. The change is not dramatic, 9.1oC ±5.1 for neat, while 4.5oC ±5.1 for 8%, the trend is apparent. The size of the endotherm does not show the same systematic trend and within the realm of statistical analysis, the size of the endotherm is invariant to cefepime concentration. At 2oC/min, the energy of the gel formation is masked by the rest of the endotherm linked with micelle formation. We are testing a lower ramp rates in order to observe the transition linked with the gel. Separately we have resolved that the gel is clearly forming and requires a cold re-equilibration to break up the gel structure. It can be inferred that cefepime is acting as a chaperone to allows micelles to nucleate more easily at lower temperature. We will present on the gel formation temperatures, enthalpies, and transitions to show the structural formation and development of the other polymers-drug complexes beyond cefepime/F-127.
8:00 PM - BM09.05.13
β-Sheet Crystallization-Driven Supramolecular Peptide Nanoagents with Structure-Dependent Theranostic Functions
Inhye Kim1,2,Eunji Lee1
Gwangju Institute of Science and Technology1,Chungnam National University2Show Abstract
Versatile, one-dimensional (1D) supramolecular theranostic nanoagents were developed by β-sheet crystallization driven-assembly of peptide amphiphiles (PAs) attached with paramagnetic metal ion (gadolinium, Gd3+)-chelating moiety and tumor cell-targeting segment, respectively. Paramagnetic metal-attached scaffolds have attracted much attention as magnetic resonance imaging (MRI) contrast agents (CAs). Many efforts have been devoted to enhance MRI efficacy of commercial CAs by adopting supramolecular scaffolds to overcome the disadvantages with low sensitivity and specificity. Here, fibrils, driven by the assembly of PA with hydrophobic β-sheet-forming peptide block, were utilized as a theranostic nanoscaffold with drug-loading within their robust core. The resulting 1D nano-aggregates allowed successful intracellular delivery of doxorubicin (DOX) to target cancer cells and contrast-enhanced MR imaging by high longitudinal (T1) relaxivity of water protons. Correlation between the structural nature of fibrils formed by PA-assembly and its diagnostic efficacy was elucidated. The nanostructured theranostics with desirable functions may thus be a useful strategy for the generation of tailor-made biocompatible nanomaterials.
8:00 PM - BM09.05.14
Graphitization and Strength of Annealed Silks and Synthetic Polymer Fibers
Thomas Dugger1,Sourangsu Sarkar2,Sandra Correa-Garhwal1,Mikhail Zhernenkov3,Hourng Kim Chea1,Cheryl Hayashi4,David Kisailus1
University of California, Riverside1,Georgia Institute of Technology2,Brookhaven National Laboratory3,American Museum of Natural History4Show Abstract
Silks have been proposed as superior carbon fiber precursors to synthetic polymers. β-sheet nanocrystals, the main contributor to silk's strength, have a favorable structure to graphitize upon annealing. Due to their uniform dispersion throughout the fiber and preferential alignment along the long axis of the fiber, silk-precursor carbon fiber could be stronger than traditional carbon fiber made from polyacrylonitrile (PAN). To investigate this theory, we compare the degree of graphitization, graphite crystal orientation, and tensile strength between spider silk, silkworm silk, PAN, and poly(vinyl alcohol).
8:00 PM - BM09.05.15
Fabrication of 2D Protein Films and 3D Protein Hollow Spheres via Alternative Self-Assembly of α-Synuclein and Their Applications
Soonkoo Lee1,Ghibom Bhak2,Seung.R Paik1
Seoul National University1,Center for Research in Biological Chemistry and Molecular Materials (CIQUS)2Show Abstract
Understanding the self-assembly process of amyloidogenic proteins is valuable not only to find their pathological implications but also to prepare protein-based biomaterials. α-Synuclein (αS), a pathological component of Parkinson’s disease, producing one-dimensional (1D) amyloid fibrils, has been employed to generate two-dimensional (2D) protein films and three-dimensional (3D) protein hollow spheres (PHS) via its alternative self-assembly at either high temperature or rapid-freezing condition, respectively. At a high temperature of 50°C, αS molecules self-assembled into the 2D film whereas 1D amyloid fibrils were produced at 37°C. This alternative self-assembly phenomenon could be attributed to structural plasticity of the intrinsically disordered protein of αS which turns into a surface active agent at the air-water interface at the high temperature. The αS 2D film was also routinely prepared at the oil-water interface and used as a framework of molecular assembly to give rise to a polydiacetylene-based sensing material. 10,12-Pentacosadiynoic acids (PCDA) were aligned on the film in a spatially organized way and then photo-polymerized to induce the π-conjugated molecular assembly yielding blue color. Its colorimetric transition to red was induced by increasing temperature. This functionalized protein film increased its height to 60 nm from 40 nm upon the PCDA immobilization and exhibited enhanced physical and chemical stability. In addition, the modified film showed remarkable high electrical conductivity only in the red state. Under frozen condition, on the other hand, PHS were produced from αS oligomers via rapid freezing, frozen incubation, and freeze-drying process. PHS prepared with αS-eosin conjugates and focused ion-beam severance of PHS confirmed their empty core structure. While PHS were stable at room temperature, they were immediately converted into amyloid burs comprised of protein nanofibrils upon heating. Therefore, PHS could be considered a constrained spherical structure transformable into biocompatible matrix material in nanoscale which could be used as a fill-in agent to improve mechanical strength of living tissues like skin as well as hydrogels in general. In this study, we have demonstrated selective fabrication of the amyloidogenic protein of αS into either 2D or 3D structures and their use as potential protein-based biomaterials.
8:00 PM - BM09.05.16
Chain-Like Superstructures of Macromolecular Micelles for Linear Assembly of Plasmonic Nanoparticles and Fluorophores
Kyungtae Kim1,Sukwoo Jang1,Jonghyuk Jeon1,Donghwi Kang1,Byeong-Hyeok Sohn1
Seoul National Univ1Show Abstract
In living organisms, we can find various chain-like structures in microscale, which are usually self-assemblies of biomacromolecules. For example, fibrils in a tissue consist of collagen as a biomacromolecular building block. Assemblies of collagens by controlled hydrogen bonding produce elongated supramolecular chains of fibrils. Similarly, colloidal nanoparticles having patches can be employed as effective building blocks for chain-like superstructures. Well-defined patches on nanoparticles can serve bonding parts for assembling process of linear chains. Especially, macromolecular micelles can have distinct patches on their surface so that they can be assembled into linear superstructures. In this presentation, we polymerized chain-like superstructures with patchy micelles of diblock copolymers and then utilized them for linear assembly of plasmonic nanoparticles and fluorophores. The growth of nanoparticles was controlled within the cores of macromolecular micelles in chain-like superstructures. In addition, fluorescent dyes were selectively attached to the core-forming blocks of macromolecular micelles to organize them into linear assemblies. We also produced red-, green-, and blue-emitting linear superstructures by varying the dyes attached to the core-forming block. We characterized optical properties of chain-like superstructures functionalized with plasmonic nanoparticles and fluorophores.
Chun-Long Chen, Pacific Northwest National Laboratory
Nico Sommerdijk, Eindhoven University of Technology
Tiffany Walsh, Deakin University
Shuguang Zhang, Massachusetts Institute of Technology
Pacific Northwest National Laboratory
BM09.06: Biomineralization and Biomimetic Crystallization
Wednesday AM, November 28, 2018
Sheraton, 2nd Floor, Back Bay A
8:00 AM - *BM09.06.01
Biomineralization-Inspired Self-Organized Organic/Inorganic Composites—Stimuli-Responsive Ordered Nanorod Materials and Aligned Thin Films Formed with Macromolecular Templates
The University of Tokyo1Show Abstract
Biomineralization-inspired synthesis of organic/inorganic composites has attracted attention. We have been developing a variety of composite materials based on macromolecular templates. Here we report hydroxyapatite and calcium carbonate nanorod composites that exhibit liquid-crystalline properties and zinc oxide thin-films with controlled aligned structures.
The TEM observations of the nanorod composites show that they have crystalline structures covered with acidic macromolecules.[2.3] These nanorods exhibit colloidal liquid-crystalline states. Hydroxyapatite nanorod materials are aligned under application of an external magnetic field. Magneto-optical response has been achieved for the liquid-crystalline states under crossed polarizers. Liquid crystalline materials are also obtained for calcite nanorods.
We have also applied bioinspired synthesis to the development of functional ZnO thin-film materials with oriented structures. We have succeeded in the biomineralization-inspired synthesis of composite thin films comprising of zinc hydroxide carbonates that are not found in biominerals. The composite thin films are converted to ZnO thin films with ordered structures through thermal treatment.
Bioinspired syntheses are useful approaches to the development of new functional materials by low-energy consumption and environmentally friendly processes.
 Kato, T.; Sakamoto, T.; Nishimura, T. MRS Bull. 2010, 35, 127; Cantaert, B.; Kuo, D.; Matsumura, S.; Nishimura, T.; Sakamoto, T.; Kato, T. ChemPlusChem 2017, 82, 107.
 Nakayama, M.; Kajiyama, S.; Kumamoto, A.; Nishimura, T.; Ikuhara, Y.; Yamato, M; Kato, T. Nature Commun. 2018, 9, 568.
 Nakayma, M.; Kajiyama, S.; Nishimura, T.; Kato, T. Chem. Sci. 2015, 6, 6230.
 Matsumura, S.; Horiguchi, Y.; Nishimura, T.; Sakai, H.; Kato, T. Chem. Eur. J. 2016, 22, 7094.
8:30 AM - BM09.06.02
Utilizing GLC-TEM to Elucidate Magnetosome Biomineralization in Magnetotactic Bacteria
Tolou Shokuhfar1,Emre Firlar1,Meagan Ouy1,Agata Bogdanowicz1,Leigha Covnot1,Boao Song1,Yash Nadkarni1,Reza Shahbazian-Yassar1
University of Illinois at Chicago1Show Abstract
Biomimicking of Fe3O4 magnetosome synthesis ex situ is of interest due to the potential uses in the physical and medical field, and is thus important to understand the biomineralization pathway for the magnetosomes in magnetotactic bacteria. Conventional TEM approaches use fixation of bacteria preventing monitoring the dynamics or using fluid cell TEM holder which does not have enough spatial resolution. Therefore, in this work, graphene liquid cells (GLC) were used to encapsulate magnetotactic bacteria after mixing them with iron rich growth medium, thus maintaining the native environment. For the first time, the formation of these nanoparticles and increased nanoparticle contrast due to advancing biomineralization using GLC-TEM was monitored.
Through electron energy loss spectroscopy (EELS) analysis, the presence of graphene optical gap, water exciton peak and graphene σ+π bond were monitored at 6 eV, 8.5 eV and at 14eV, respectively. Formation of radiation induced H2 bubbles and magnetosomes were observed as well, indicating the presence of liquid during electron imaging. Fe2+ (octahedral), Fe3+ (tetrahedral), Fe3+ (octahedral) and FeOOH reference spectra were used to fit the experimental data. Relative ratio of Fe2+ to Fe3+ was calculated to be 0.35. Magnetite was known to be able to be converted to first maghemite and then to hematite via the electron beam exposure. Considering this ratio will be 0.5 for a perfect Fe3O4 (1x Fe2+, 2x Fe3+), hematite is the strongest candidate to be present in addition to magnetite in magnetosomes contributing to higher Fe3+ content. At different time scales after iron induction, (i) an increase of the magnetosome image contrast was reported by the line profile drawn across the magnetosome and increased TEM contrast showed increased mass-thickness in the image, which indicates progression in biomineralization and incorporation of more Fe3O4 molecules to this particle; and (ii) an increase in the number of magnetosomes were observed.
8:45 AM - BM09.06.03
The Formation of Macromolecular Silica Nanocomposites Through Self-Assembly and Biomineralization
Paula Vena1,Demi de Moor1,Heiner Friedrich1,Joseph Patterson1,Nico Sommerdijk1
TU Eindhoven1Show Abstract
In nature we can find many examples of organic-inorganic nanocomposites with exceptional properties. For example, diatoms are unicellular algae whose cell walls are formed form hierarchical nanostructured silica. Euplectella sp. is a siliceous sponge that can assemble a mechanically resistant glass cage with nano- to macro- hierarchical organization. In both cases the formation process occurs by the intricate interactions between macromolecules and the silica precursors. These beautiful examples provide inspiration for the controlled formation of structurally complex silica based materials under ambient conditions.
In this paper we present a detailed study on the use of synthetic macromolecules to control silica morphology using two strategies: 1) mineralization of silica in pre-assembly macromolecular templates with defined pores and 2) the co-assembly of silica precursors and nanoparticles with macromolecules. Using (cryo)-electron microscopy, (cryo)-electron tomography and liquid phase electron microscopy we investigate the formation processes and characterize the hierarchically porous silica structures.
We discuss the underlying principles of silica mineralization in confinement and design strategies for making hierarchically porous silica based materials. Such porous materials have many potential applications including insulators, sensors, catalysts and drug delivery vehicles. We hope the insights provided by our detailed investigation will help towards the rational design and understanding of mechanisms formation of porous silica based materials.
9:00 AM - BM09.06.04
Mineralization in Bio-Inspired Metal-Coordinate Polymer Hydrogels
Massachusetts Institute of Technology1Show Abstract
Biominerals have been widely studied in part due to their unique mechanical properties, afforded by their inorganic-organic composite structure and well-controlled growth in macromolecular environments. More recently, growing concerns over climate change and environmental sustainability and the emerging relevance of green chemistry make biomineralization an even more attractive process to study. Here, we focus on the earlier stages of mineral nucleation and growth in macromolecular environments, where an organic, hydrogel matrix dominates the bulk properties of the material and the mineral is distributed throughout the matrix as nano- and/or microparticles. The phase, morphology, and size of the particles can be controlled using the choice of the hydrogel, functional moieties on the gel polymer backbone and soluble additives. Depending on the choice of organic matrix and inorganic mineral, the matrix can be dissolved to leave highly uniform particles, or the matrix can be left intact, creating a hydrogel-mineral composite with improved mechanical properties through organic-inorganic interfacial interactions or additional functionality, such as magnetic properties.
9:15 AM - BM09.06.05
Towards Templating 2D Magnetite Platelets via Bio-Inspired Approaches
Bernette Oosterlaken1,2,Giulia Mirabello1,2,Yifei Xu1,2,Joseph Patterson1,2,Heiner Friedrich1,2,Nico Sommerdijk1,2
Eindhoven University of Technology1,Institute for Complex Molecular Systems2Show Abstract
Magnetite, Fe3O4, is a naturally occurring iron oxide. Magnetite has excellent mechanical properties, as well as magnetic properties. Its magnetic properties depend on crystal size and shape. In nature, the formation of magnetite is precisely regulated, as controlled size and shape crystals are specifically tuned depending on the biological function, even at ambient and aqueous conditions.
Inspiration for this work was found in nature, where specialized vesicles with associated transmembrane proteins are directing nucleation and growth of the magnetite crystals. Achieving a similar level of control over crystal shape and size thus far has been challenging in synthetic procedures and the processes behind templated magnetite mineralization are still poorly understood.
Little work has been done on templated magnetite formation so far. In a bio-inspired approach, we are investigating templated magnetite growth, to precisely tune the size and shape of the magnetite crystal into a certain shape. The targeted crystal shape in this project is 2D platelets. 2D platelets of magnetite might have appealing magnetic properties, such as magnetic vortices, which result in in-plane magnetization.
A suitable template to direct magnetite growth into 2D crystals is collagen. Collagen is known to template the formation of calcium phosphate into 2D platelets, but also lepidocrocite (γ-FeOOH) (Xu et al., in preparation, 2018). By growing magnetite into a collagen template, we are creating a novel collagen-based hybrid material with excellent mechanical and magnetic properties. To the best of our knowledge, we are the first to explore the possibilities of growing magnetite in a collagen template.
To template magnetite inside a collagen matrix, magnetite formation outside the collagen template should be inhibited. Acidic (bio-)molecules, like polypeptides, are shown to influence magnetite crystal growth. As such, those polypeptides are used to assist in mineral formation inside the template, similar to what has been exploited for calcium phosphate mineralization in a collagen matrix before.
Combining spectroscopic techniques such as Raman spectroscopy with advanced electron microscopy techniques might provide us with new insights in the mechanisms behind magnetite formation inside the template. CryoTEM already has been shown to be of great value when addressing mineral formation mechanisms. Liquid phase electron microscopy (LP-EM) allows to visualize the processes in-situ and therefore is an appealing complementary technique to CryoTEM. Preliminary experiments in the confined space of the liquid cell that is used for LP-EM measurements show that magnetite indeed has formed inside the liquid cell. After optimization of the experimental conditions, LP-EM allows for in-situ visualization of templated magnetite formation.
10:00 AM - *BM09.06.06
Crystallization in Confinement—A Bio-Inspired Approach
Fiona Meldrum1,Clara Anduix-Canto1,Yun-Wei Wang1,Yi-Yeoun Kim1,Shunbo Li2,Hugo Christenson1
University of Leeds1,Chongqing University2Show Abstract
The organisation and function of biological systems is based on compartmentalisation. Biomineralisation processes, which lead to the generation of mineral-based structures such as bones, teeth and seashells are no exception to this. Biominerals form within the confines of “privileged environments” delineated from the organism, where spatial constraints and chemical conditions can be precisely controlled. Despite this, the influence of confinement on crystallisation processes is poorly understood. This talk describes a series of systematic investigations into the effects of confinement on the formation of a range of important crystal systems including calcium carbonate, calcium sulfate and calcium phosphate. Rods of controlled pore glasses (CPGs) with sponge-like structures were used to access true nanoscale confinement. X-ray absorption and diffraction tomography were used to study the precipitation of a population of calcium sulfate particles within these environments and we show that bassanite (CaSO4.0.5H2O) – whose existence as a precursor to gypsum (CaSO4.2H2O) has been disputed – is stable in the CPGs for periods of at least three weeks. We can also monitor the transformation from bassanite to gypsum and investigate how this leads to fracture of these porous media. At larger length scales, microfluidic devices including a novel “crystal hotel” that comprises a series of “rooms”, are used to grow larger crystals. These offer many features including flow, confinement and the possibility of changing reaction conditions with time that are common to biological systems. These devices were used to image the development of crystals within confined volumes and to study the effects of soluble additives on calcium carbonate precipitation at early times. Our results show that the additives have no effect on crystal morphologies until the crystals reach at least 100 nm in size. Finally, the cylindrical pores of track-etched membranes were used to study the effects of confinement on calcium carbonate and calcium phosphate precipitation. We show that the polymorph of CaCO3 changes from calcite to aragonite as the pore diameter decreases, and that aragonite is the major polymorph in 200 nm pores when low concentrations of magnesium and sulfate ions are also present. Finally, calcium phosphate precipitation in this system generates highly oriented single crystal of hydroxyapatite in small pores, where the degree of orientation is comparable to that seen in bone. Confinement therefore enables effective control over crystallisation processes, and as such promises the ability to optimise the synthesis of crystalline materials.
10:30 AM - *BM09.06.07
The Invertebrate Calcium Carbonate Biomineralization Process is Guided by Protein Hydrogels
New York University1Show Abstract
In the development of the invertebrate calcium carbonate skeletal elements, protein families are involved in the coordination of mineralization events. In many instances this process begins with the formation of amorphous calcium carbonate mineral nanoparticles which are assembled into mesoscale crystals (calcite, aragonite, vaterite) and this assembly process creates many mechanistic features of these crystals that are important for material integrity. We have observed that in the mollusk shell and sea urchin spicules that this mineralization process is guided by the formation of matrix protein hydrogel phases that stabilize, assemble, and organize mineral nanoparticles, create nanotextured crystal surfaces, and form intracrystalline nanoinclusions within the mineral phase that introduce elastic deformability to the mineral phase. These protein hydrogels form in response to protein-protein interactions that are entropically driven by intrinsically disordered and modified globular protein domain sequences, as well as protein-carbohydrate interactions involving glycoproteins. We hope that materials science will develop similar principles to create the next generation of composite materials under ambient conditions.
11:00 AM - BM09.06.08
Coding Cell Micropatterns through Peptide Inkjet Printing for Arbitrary Biomineralized Architectures
Wenyi Li1,Jin Guo1,Shengjie Ling2,Ying Chen1,Chunmei Li1,Fiorenzo Omenetto1,David Kaplan1
Tufts University1,ShanghaiTech University2Show Abstract
Well-designed micropatterns present in native tissues and organs involve changes in extracellular matrix compositions, cell types and mechanical properties to reflect complex biological functions. However, mimicking these micropatterns in vitro remains a challenge and the patterning strategies often showed limited guidance of cell orientation in relatively short culture periods. Silica-based micropatterns are popular in many biomedical fields including in vitro tissue models, due to the biocompatibility and high versatility of silica. Yet, harsh conditions (e.g. extremely high temperature and/or pressures) are often required to create silica patterns, and bonding between substrates is not strong enough. In this work, a de novo design strategy to code functional micropatterns to engineer cell alignment through the integration of aqueous-peptide inkjet printing and site-specific biomineralization is presented. Inkjet printing provides direct writing of macroscopic biosilica selective peptide-R5 patterns, which allow site-specific growth of silica nanoparticles through in situ biomineralization, with micrometer-scale resolution on the surface of a biopolymer (silk) hydrogel to achieve the alignment of human mesenchymal stem cell (hMSCs) and enhanced immobilization of bovine serum albumin (BSA).
To create the micropatterns, peptide-R5 was inkjet printed on the surface of enzymatically crosslinked silk hydrogels, followed by subsequent silicification to induce biosilica deposition. Linewidth and gap distance between each printed line were manipulated by adjusting drop spacing and drop volume during printing. Biomineralization was confirmed by examining silica nanoparticles covering the printed lines but not elsewhere. A 20 µm pattern gap distance and 1 µm linewidth were achieved. Well-defined peptide patterns on the substrate were also evidenced by printing fluorescein isothiocyanate (FITC)-labeled R5 and observed by fluorescence microscopy. hMSCs were cultured on the micropatterned hydrogels and specific alignment along the printed lines was noted, while the response on the unpatterned controls was randomized. Additionally, FTIC-labeled BSA and R5 were printed together and after 6 days of incubation in phosphate buffered solutions (PBS), BSA immobilized and aligned exclusively along the biosilica micropatterns, while the FITC-BSA alone extended over the surface, which suggests improved protein stability and alignment on micropatterns.
A de novo strategy to design functional micropatterns to engineer cell alignment and protein immobilization through inkjet printing and site-specific biomineralization was demonstrated. This cost-effective micropattern design scheme can meet a wide range of needs in the biomedical field with implications for broader material designs.
11:15 AM - BM09.06.09
Bio-Inspired Approaches to Creating Functional Nanocomposite Crystals
Yi-Yeoun Kim1,Alex Kulak1,Fiona Meldrum1
University of Leeds1Show Abstract
The production of crystalline materials with structures and properties resembling those of biominerals is a challenging synthetic goal. Biominerals are invariably composite materials in which organic matrices are associated with the inorganic phase, even single crystal biominerals contain proteins embedded within the crystal lattice. Biominerals therefore provide a unique inspiration for the design and synthesis of new materials.
This talk explores how this biogenic strategy can be used to generate synthetic crystals with novel composite structures and properties and to determine the “design rules” which govern the occlusion of additives within crystals. Using polymer particles rather than proteins as simple crystal growth additives, High levels of particle occlusion is achieved by tuning the particle surface chemistry and the crystal growth conditions within calcite single crystals.
Our strategy is extended to generate nanocomposites in which inorganic nanoparticles are distributed throughout a crystal matrix with true nano-scale mixing. Highly effective incorporation of gold and magnetite nanoparticles was achieved within host carbonate- and sulfate- minerals by controlling the nanoparticle surface chemistry using a physically-adsorbed double hydrophilic diblock copolymers. This methodology can potentially be applied to a huge number of nanoparticle/ host crystal systems, where its experimental simplicity makes it an attractive and general method for generating composite materials.
11:30 AM - *BM09.06.10
Exploring the Abiotic-Biotic Interface—From Fundamentals to Biomimetic Composites
Nottingham Trent Univ1Show Abstract
Events occurring at the solid/aqueous interface (i.e. molecular recognition, adsorption, desorption etc.) underpin a variety of technologies used in the biomedical and biotechnological fields. The use of nanoparticles and multifunctional nanoparticles that combine recognition and targeting with specific properties is widespread for the development of clinical diagnostic tools or therapeutic platforms. Although we are developing understanding of how materials are made in nature there is a long way to go in applying that knowledge in a systematic and predictive fashion to develop new composite materials.
This presentation will highlight (a) aspects of our recent exploration of the effect of surface chemistry on peptide material interactions using both simulation (DFT) and experiment, and (b) show how we can apply this understanding to the development of novel biomimetic composites based on Pt/Au and MOFs.
In all our studies we use a wide range of experimental techniques to characterise our (bio) materials and measure their behaviour at interfaces. The research presented will include new methods being developed specifically to probe such interactions.
BM09.07: Biomimetic Crystallization
Wednesday PM, November 28, 2018
Sheraton, 2nd Floor, Back Bay A
1:30 PM - *BM09.07.01
Creation of Hierarchical Material Structures Using Molecular Specificity
University of California, Los Angeles1Show Abstract
Material formation in nature is precisely controlled in all aspects from crystal nucleation, growth to assembly to deliver superior functions. Specific biomolecule-material interactions have been hypothesized to play important roles in these processes. Proteins, polymers and small molecules have been extensively explored to replicate the degree of control in material formation in vitro and for nonbiogenic materials. However the organic-inorganic interfacial interaction is still far from being understood which hinders the further advancement of biomimetic material formation. In this talk I will share our efforts on decoding the myth of biomolecular specificity to material surface and their roles in controlling crystal nucleation and growth. The selection of facet specific short peptides and their abilities in guiding predictable morphology control of Pt nanocrystals will be first demonstrated. Then detailed experimental and theoretical studies on binding mechanism will be discussed. These studies open up opportunities in understanding the molecular details of inorganic-organic interface interaction, which can one day lead to the development of a library of molecular functions for biomimetic materials design and engineering.
2:00 PM - *BM09.07.02
Leveraging Molecular-Level Control of Peptide Constructs to Direct the Synthesis, Structure and Properties of Chiral Nanoparticle Superstructures
University of Pittsburgh1Show Abstract
Replacing one atom or linkage in an organic molecule or polymer can dramatically affect its structure and properties. Chemists have leveraged the power of synthesis to adjust and fine tune the properties of molecules. Nanoparticles are a class of fundamental structural and functional building blocks for the construction of new materials. The properties of these materials depend intrinsically on the size, shape, and composition of the constituent nanoparticles as well as the precise organization of the nanoparticles within the material. In order to fine tune the properties of the material, we must be able to carefully adjust the organization of its component nanoparticles. Can we use the power of synthetic chemistry to program and carefully adjust the structure and properties of hierarchical nanoparticle-based materials? This talk deals with peptide-based methods for controlling the synthesis and assembly of nanoparticles into well-defined chiral helical architectures. Rigorous molecular models of peptide assemblies will be detailed. It will be demonstrated that the atomic make-up of the peptide constructs can be carefully adjusted and that these subtle yet purposeful modifications lead to non-trivial structural changes to the chiral nanoparticle superstructure assembly and properties.
3:30 PM - *BM09.07.03
Engineering Biology for Design and Assembly of Functional Materials
Steve Kim1,Rajesh Naik1,Joseph Slocik1,Kristi Singh1,Maneesh Gupta1,Kuang Zhifeng1,Patrick Dennis1
Air Force Research Laboratory1Show Abstract
Biological systems offers inspiration and exciting opportunities for creating biomimetic materials, structures and devices. The assembly of individual biomolecular units into well-defined, higher-order functional structures is a hallmark of biological systems, and is exemplified in the self-organization of biological building blocks into supramolecular structures (e.g. peptides, proteins, nucleic acids, viruses). Such biological materials/systems offer inspiration and exciting opportunities for creating biomimetic materials, emulating processes and inspiring the design of devices. Furthermore, the ability to use synthetic biology tools to manipulate the genetic information encoding for biomolecules of interest allows one to design materials with tailored functionalities and properties. I will describe in my talk our research on engineering biology to create designer biomolecules or enabling the assembly of functional materials for various applications.
4:00 PM - BM09.07.04
Investigations into the Mechanism of Biosilicification Under In Vitro Conditions
Sai Maddala1,Ernst van Eck2,Paul Bomans1,Heiner Friedrich1,Nico Sommerdijk1
TU Eindhoven1,Radboud University2Show Abstract
Diatom biosilicas possess intricate organization, whilst requiring only ambient synthesis conditions. Their remarkable morphological properties have been a source of intense interest for chemists and biologists. Understanding the mechanisms involved in the morphological control of biosilica could inspire the production of new functional materials.
Here, we present results from our in vitro investigations into biomineralization of silica. Our investigations focus on mimicking the conditions present in the silica deposition vesicle of the diatoms; high silicic acid concentration (0.10 to 0.34 M), salinity, the presence of polyamines and mildly acidic pH (pH 5.5). Understanding the silica formation in these conditions is key to unravelling the mechanism of silica biomineralization.
Tetramethyl orthosilicate (TMOS) was used as silica precursor, as it readily hydrolyses to give metastable silicic acid solutions. pH was maintained at 5.5 using an autotitrator. Silicic acid consumption was monitored using ammonium molybdate assay. In the absence of polyamine additives, and under high salt conditions (NaCl concentration 0.45 M) free silicic acid was consumed in two stages, first a portion of it undergoes rapid condensation within 5 minutes, and the remainder stays unreacted for the next 27 minutes, followed by complete reduction in concentration by 43 minutes. Previous investigations into silica formation were generally performed in pure water, and this two-stage silicic acid consumption process was not observed. The role of polyamines on silica particle formation was monitored under real-time conditions using Dynamic Light Scattering (DLS). We used polyallylamine hydrochloride (PAH, Mw 15000 g/mol) as polyamine mimic. In the absence of PAH, silica particles were observed as soon as TMOS completely hydrolysed in water. In the presence of PAH, the particle formation wasn’t observed for the first 76 minutes, suggesting that the polyamines could potentially inhibit silica formation. Argon sorption porosimetry of silica gels obtained in the absence of PAH had a surface area of 170 m2/g and a pore size of 7.05 nm. Whereas in the presence of PAH, the surface area was 230 m2/g and pore size of 3.36 nm. Further analysis was performed using Cryo-TEM and solid state 29Si NMR.
Our results indicate that under the conditions found in silica deposition vesicle of diatoms, silicic acid undergoes rapid condensation and that the presence of polyamines retard this process. This suggests that the polyamines could crucially help stabilize and store silicic acid. While the exact mechanism will be the subject of future research, our results help explain an important intermediate step in silica biomineralization.
4:15 PM - BM09.07.05
Building Biomimetic Bone at Higher Level of Organization
Elora Bessot1,2,Clement Sanchez3,Marco Faustini1,2,Nadine Nassif4,1,5
UMR 7574 - Laboratoire de Chimie de la Matière Condensée de Paris1,Sorbonne Université2,Collège de France3,CNRS4,ESPCI5Show Abstract
Bone is a composite material which closely associates a dense and organized collagen organic matrix (mainly type I collagen fibrils) with an apatite mineral network. From nanometers to millimeters and beyond, bone is hierarchically structured to provide maximum strength with a minimum of material. The structure/function relationship being crucial in bone, attention needs to be paid to the long-range collagen/hydroxyapatite (HA) structure in models set in vitro. Recently, the cholesteric geometry was reproduced in the laboratory (Wang and al., Nat Mater 2012).
The aim of this work is to reach higher levels of bone hierarchical organizations enlarging the relevance and applications of bone models. We are working on two different organizations : the trabecular and the cylindrical motif (osteons) of the cortical bone. Both imply the texturization of the liquid-crystalline phase made of a mixture of highly concentrated acidic collagen with the HA precursors. For this purpose, different physical constraints are applied to collagen liquid-crystal phases to control the spatial arrangement of the oriented domains. Thanks to the geometry, the direction of the flow or/and the confinement, the shear forces involved may have an effect on the resulting organization.
The resulting hybrid biomimetic materials and their hierarchical organization require various characterization techniques (e.g. in-situ observations by polarized optical microscopy (birefringence) and investigation by SAXS/WAXS of the directed co-assembly of the organic/inorganic phases, electronic microscopies, mechanical tests etc).
By using an original strategy, we aim to improve our knowledge on processes involved in the bone tissue (morphogenesis) as well as build new biomimetic hierarchically-structured materials that offers remarkable scaffolds to repair larger defects for bone tissue engineering.
4:30 PM - *BM09.07.06
Harnessing the Precision of Biorecognition for the Development and Assembly of Responsive, Functional Inorganic Nanomaterials
Univ of Miami1Show Abstract
Nature has exploited the precision of biorecognition events for the development inorganic materials for critical applications ranging from protection against predation to structural support. These materials are generated under sustainable conditions where the translation of such approaches to material compositions of technological importance could provide pathways to address current needs in applications ranging from energy harvesting and storage to biological sensors and theranostic systems. At present, only minimal understanding is known concerning the direct interaction between biological and bio-inspired molecules (e.g. peptides, DNA, peptoids etc.) with inorganic materials, where the ability to predictably design these biomolecules with affinity for the target system remains unachieved. By having such capabilities, the ability to fabricate functional materials with desired properties on demand could be accessed for immediate use in targeted applications. In addition, due to the great complexity achievable from biosystems, the biomolecules could be designed with secondary functionalities beyond inorganic material affinity, thus generating final structures with multifunctional capabilities. Our research has focused on the design of new bio-inspired systems with the ability to fabricate functional inorganic materials and drive their assembly in three dimensions. This assembly process is accessed based upon the multifunctional capability of the peptides to recognize and bind the inorganic surface, while simultaneously self-organizing in three dimensional space. In one instance, the self-assembly process is driven through biomolecule-biomolecule interactions, while in a second case, the assembly process is achieved through crosslinking of multiple inorganic materials from a single biomolecule. This research demonstrates multiple, disparate pathways from which biorecognition events can been exploited to drive inorganic material assembly, which could be tailored to different systems based upon biomolecular affinity.
BM09.08: Poster Session II: Bioinspired Materials
Thursday AM, November 29, 2018
Hynes, Level 1, Hall B
8:00 PM - BM09.08.01
Dehydration Stability Analysis of DNA-Guided Nanoparticle Superlattices
Hayato Sumi1,Takumi Isogai1,Shoko Kojima1,Shunta Harada1,Toru Ujihara1,Miho Tagawa1
Nagoya University1Show Abstract
Nanometer-scale materials which have unique electric, photonic, phononic and magnetic properties are hard to control and create. The self-assembly of DNA-guided nanoparticles has attracted much attention as a novel technique to design nanostructures flexibly due to the programmability of DNA base sequences. DNA-functionalized nanoparticles (DNA-NPs) can assemble into various types of 3D nanoparticle superlattices by using sequence-selective DNA hybridizations. However, DNA-NP superlattices have a problem in structure stability. Whereas DNA-NP superlattices are stable in a buffer solution, they collapse outside the solution because of the dehydration of DNA strands. Resin- or silica-based encapsulation method is one way of solving the problem in terms of the stabilization. For a wide range of applications, the direct dehydration of DNA-NP superlattices, without any filler, is crucially important. It has been reported that the volume fraction of nanoparticles per DNA-NP superlattice volume is relatively sensitive to the structure stability of the DNA-NP superlattice during dehydration, however the optimum condition of the volume fraction for the structure integrity has not yet been studied. Here, we investigate the optimum condition for the dehydration of DNA-NP superlattices by controlling the volume fraction of nanoparticles per unit cell in solution by using different sized nanoparticles.
Firstly, Au nanoparticles were functionalized with thiolated-DNA strands. These DNA-functionalized nanoparticles were combined with linker DNA strands, which hybridize to complementary sequences. Secondly, the mixture solution was heated up to 65 oC and then slowly cooled back to 25 oC. By changing the combination of DNA base sequences and nanoparticle sizes, we designed 3D superlattices to be assembled into bcc, fcc and CsCl structure, respectively. The crystal structures of DNA-NP superlattices were analyzed by small angle X-ray scattering (SAXS) before and after dehydrations. The shapes and the surface structures of dehydrated samples were observed by scanning electron microscopy (SEM).
By analyzing SAXS patterns, the assemblies of DNA-NPs in solution were identified as bcc, fcc and CsCl superlattices as we designed. We also tried to analyze the assemblies of DNA-NPs after dehydration and for the first time we have succeeded in highly accurate structure analysis of direct dehydrated DNA-NP superlattices with higher volume fractions of nanoparticles per unit cell, which exhibited clear diffraction patterns. It has also been confirmed that direct dehydrated DNA-NP superlattices with higher volume fractions tended to maintain their lattice symmetries regardless of crystal structures. SEM analysis confirmed DNA-NP superlattices with faceted crystal shapes and ordered arrangements of Au nanoparticles on their crystal faces at higher volume fractions, which also indicates the structurally stable dehydration process while retaining their lattice symmetries.
8:00 PM - BM09.08.03
Catalyzed Organic Reactions in Aqueous Media Using Hierarchically-Structured Nanomaterials Assembled from Sequence-Defined Peptoids
Tengyue Jian1,Chun-Long Chen1
Pacific Northwest National Laboratory1Show Abstract
Natural enzymes are highly efficient and selective catalysts that present unique catalytic microenvironments. Development of highly stable enzyme- mimic catalysts using sequence-defined synthetic molecules will benefit the area of biomimetic catalysis and facilitate our understanding of the structure-depended catalytic performance. Here we report the design and synthesis of peptoid-based biomimetic materials with hierarchical structures for catalyzing the direct asymmetric aldol reaction and hydrolysis reaction in aqueous media. First we synthesized and assembled 20 proline-containing membrane catalysts, and demonstrated their catalysis of asymmetric aldol reaction with high conversion yields and good enantio- and diastereoselectivities. We further showed that both the enantio- and diastereo-selectivities of this aldol reaction are highly dependent on the hydrophobic microenvironment built around catalytic sites. For the hydrolysis reaction, we found that imidazole and pyridine-containing 2D membranes catalyzed this reaction in aqueous media with high efficiency. We demonstrated the use of peptoid nanomembranes for degradation of nerve agent simulants: 4-nitrophenyl phosphate and acyl ester (nitrophenylacetate and pyrene ester). In this catalyzed hydrolysis reaction, we found that the addition of Zn2+ and Cu2+ significantly accelerated the hydrolysis rate. We further demonstrated that these catalyzed organic reactions are highly dependent on the morphology and crystallinity of peptoid assemblies.
8:00 PM - BM09.08.04
Ultrastable Supramolecular Hydrogel of Hydrophobic Peptides Prepared by a Hydrolysis Process
Nankai University1Show Abstract
Supramolecular hydrogel based on peptides self-assembly have attracted extensive research interests in recent years, but its application in the field of nanomedicine is still limited by its poor stability in extreme environments. The number of hydrophobic amino acids directly affects the solubility of the peptides and the properties of the hydrogel. Peptides FF (F: Phenylalanine) has strong assembly capability, those hydrogels based on dipeptide of FF are the most widely investigated. Organic solvent is always required to dissolve peptides because of its strong hydrophobic properties. However, there is no hydrogel containing more than three Phenylalanine currently in aqueous solution.
TTherefore, we describes a method for the preparation of hydrogels containing hydrophobic FFFF and FFFFF sequences by ester bond hydrolysis, which resulted in two highly potent peptide hydrogels formed by Nap-GFFFFF-GP-EE and Nap-GFFFF-GP-EE (GP: Glycolic Acid). Then the transparent hydrogels were formed at 37 degrees Celsius for about 48h. The hydrogel had excellent stability in highly acidic (pH=1) and highly basic solutions (pH=13) for 15 days.
In conclusion, a novel method was constructed to promote hydrophobic peptides to dissolve in water and form a supramolecular hydrogel by the ester and the bond hydrolysis method, even if the molecule itself is unable to form a hydrogel and there is low solubility with ultrasound or heat means. This approach should be applicable for exploring supramolecular assemblies formed by other hydrophobic peptides. From the above, useful information for hydrogels of hydrophobic peptides has been presented through our study.
8:00 PM - BM09.08.06
Fine-Tuning Core-Shell Nanoparticle Growth by Exploiting Ions Doping
Jianxiong Zhao1,Xian Chen1,Bing Chen1,Feng Wang1
City University of Hong Kong1Show Abstract
Predicting and tailoring the morphology of core shell nanoparticle is indispensable in obtaining desirable properties for optical and biological applications. However, the incomplete and discontinuous shell growth induced by heterogeneity in structure and chemical bonding may reduce the luminescence intensity and detriment to particle uniformity. Here, we demonstrate a strategy of ions doping for forming continuum NaREF4 (RE= rare earth) shell growth on heterogeneous core.
Compared to those un-doped shell which formed homogeneous nucleation surround the core. We show that doping can greatly reduce the energy barrier of heteronucleation, making shell successfully deposit on core. The core shell structure are distinguished by using high-angle annular dark-field scanning transmission electron microscopy. The shell growth process at different reaction stages are revealed by transmission electron microscopy observation on nanoparticle extracted during the reaction. We show that nascent shell layer formed at low energy crystallographic facets, afterward, shell extended and merged together over entire core surface.
We applied the strategy on core particle with different size and dimensions. All shell growth is stabilized but with some morphology variation for different core shape. The TEM analysis show that shell growth is not only strongly dependent on the nature of the different facet, but also reliant on mechanical flexibility of core. Moreover, the strategy improves both upconversion emission and lifetime of core shell nanoparticle and simultaneously provide novel platforms for building multifunctional self assembled composites.
8:00 PM - BM09.08.07
Liquid Crystalline Disubstituted Polyacetylene-Preparation, Properties and Possible Applications for Biomimetic Materials
Kyoka Komaba1,Masashi Otaki1,Hiromasa Goto1
University of Tsukuba1Show Abstract
Many polymers mimicking ecosystem such as DNA have been synthesized. Synthesis of helical macromolecules has been inspired by such natural polymers. In this report, we synthesized disubstituted polyacetylenes using tantalum based catalyst. Chemical structure and optical properties of the polymer thus synthesized in this study are characterized with infrared absorption spectroscopy (IR), gel permeation chromatography(GPC), UV-vis absorption (UV) spectroscopy, circular dichroism (CD), fluorescence spectroscopy, and polarized optical microscopy analyses. Polyacetylene as a conductive polymer has some drawbacks such as poor stability in the air, no solubility, low processability. Introduction of substituent to the polyene backbone allows improvement of its disadvantages. Introduction of liquid crystal group can improve such drawbacks. We synthesized liquid crystal polyacetylenes for potential applications in biological field.
We succeeded in synthesis of disubstituted polyacetylene using tantalum chloride catalyst (Y = 41%). GPC result indicated that number average molecular weight (Mn) of the polymer is to be > 1,000,000 and degree of dispersion (Mw/Mn) ca.1.4.
The IR spectroscopy measurements confirmed that polyene was successfully produced by opening of the C≡C bond of the monomer in the polymerization. Polymer shows two absorption maxima at around 370 nm and 430 nm in the UV-vis. These absorption bands are derived from p-p* transition of the main chain. The polymer shows no CD signals. This result indicates that the polyacetylene derivative synthesized in this report contains equimolar amount of right-handed and left-handed helical structures, resulting in racemic state. This polymer shows fluorescence signal at around wavelength 500 nm.
Polarizing optical microscopy observations for the drop cast film from toluene solution deposited on a glass substrate was carried out. This polymer shows fan-shaped texture of smectic liquid crystal with lyotropic liquid crystallinity. The layer structure has similarity to biological membrane. This unique structure may be artificial lipid bilayer membrane.
In this work, we achieved synthesis of disubstituted polyacetylenes with large molecular weights, showing lyotropic liquid crystallinity with layer structure. We evaluated that the simple synthesis of liquid crystal poltyacetylene derivatives having no mesogenic group from readily available monomers affords to production of the biomimetic lipid bilayer membrane.
8:00 PM - BM09.08.08
Selenium- and Tellurium-Containing Block Copolymer with Multi-Hierarchical Oxidation Response
Lu Wang1,Huaping Xu1
Tsinghua University1Show Abstract
Nanomaterials with hierarchical responsiveness are of great significance for not only fundamental science but also future biomedical applications due to sophisticated and hierarchical physiological environments. Here, we report a selenium- and tellurium-containing block copolymer that can be stepwise oxidized by both chemical methods and electrochemical methods. Differences in sensitivity to the oxidation of selenium and tellurium were employed. By tuning the concentration of the oxidant and oxidation periods, self-assembly behaviors of the copolymer were tuned by stepwise chemical oxidation. After oxidation, some interesting morphological evolution was observed that the polymer micelles crosslinked with each other without any swelling. In the case of electrochemical oxidation, the voltage during the electrochemical oxidation and oxidation period also affected the level of oxidation. Furthermore, we showed that the degree of electrochemical oxidation varied with a different PEG block length. Considering sophisticated physiological conditions in vivo, this hierarchically responsive system may provide new possibilities as smart delivery vehicles in biological environments.
8:00 PM - BM09.08.09
Sequence-Designed pH-Responsive Pure DNA Hydrogel Produced by Rolling Circle Amplification
Guoyuan Liu1,Leilei Tian1,Yishun Huang1,Wanlin Xu1,2,Haoran Zhao1,Pan Li1,Jing Li1
Southern University of Science and Technology1,Zhengzhou University2Show Abstract
Stimuli-responsive DNA hydrogels, albeit the potential advantages like biocompatibility and ease to functionalize, problems remain in the reconciliation between facile fabrication and stimuli-responsiveness. In this study, by rationally designing the sequence of DNA chains, pH-responsive sites-namely, i-motif forming sequences (IFSs)- were introduced during rolling circle amplification. At pH 5.0, the resultant gelation occurs driven by the formation of intermolecular i-motifs as crosslinkers.
To better evaluate the sequence influence on the responsiveness, three IFSs were designed and named as I1, I2, and I3 whose sequences are CCCCCTCCCCC, CCCTCCCTCCCT, and CCCAATCCCAATCCCAATCCC. Three IFSs form primarily inter-molecule, both inter-and intra-molecule and primarily intra-molecule i-motif structure respectively. The difference in sequence reflects on the pH-responsiveness during sol-gel transition. When adjusting the pH from 5.0 to 8.0, I3 quickly dissolved while I1 became softer but remain intact and I2 became more dispersed in the buffer. We draw a conclusion that the gelation process can be tuned via sequence design. An IFS favoring intramolecular i-motif leads to mechanically weak, thermally unstable yet pH-sensitive hydrogel while for IFS favoring intermolecular i-motif structure, gelation may occur along with quick condensation which promotes non-specific interactions within the gel.
The microstructure of the hydrogel was investigated to dig some evidence of the gelation mechanisms. RCA products at neutral (pH 7.5) and acidic condition (pH 5.0) were observed under scanning electron microscope. Of all three samples, flower-like microstructures (FMs) with diameter of 2-3 μm was observed in a large amount. FMs were believed to be a in situ formed by-product of DNA polymerization and performs a crucial role in gelation process. Form the SEM images, the hydrogel morphology featured with thick DNA matrix embedded with FMs. For sol, the FMs is more visible due to the lack of matrix surrounding. Also, the different properties of specimen can be explained. For I1, dense matrix already formed in neutral pH indicating many crosslinking sites that are not i-motif structures. For I3 the matrix in acidic buffer is relatively looser which is consistent with the weak stability. Worth noting that there is no RCA matrix in I2 after redissolved in pH 8.0 buffer yet the FMs remain unchanged. This proves that FM, as a condensed complex of DNA and magnesium pyrophosphate (MgPPi), has no pH-responsiveness and that the pH-adjusting gelation exclusively relies on introduced i-motif sequences.
To summarize, this work proposed a facile method to produce pH-responsive DNA hydrogel. Also, the influence of three different IFS sequences on gelation mechanism as well as their interaction with flower-like microstructures were investigated.
8:00 PM - BM09.08.10
Hybrid Thin-Film Formation of Zinc Layered Hydroxides with Intercalated Organic Molecules Through a Biomineralization-Inspired Approach
Satoshi Kajiyama1,Takashi Kato1,Fumiya Katase1
Univ of Tokyo1Show Abstract
Biomineralization-inspired crystallization is one of effective approaches for the development of hierarchical structures from nano- to macro- scales under ambient conditions [1-3]. In biomineralization, acidic polymers induce amorphous states of inorganic crystals with the interaction between acidic groups and metal ions. The amorphous states are useful as precursors for inorganic crystals with ordered structures [4,5]. We have achieved the formation of hybrid thin films based on zinc hydroxide carbonate (ZHC) with ordered nanostructures through biomineralization-inspired approach utilizing amorphous precursors. ZnO thin films have been obtained with ordered structures from ZHC ordered hybrid thin films through thermal treatment .
In the present study, we demonstrate that hybrid thin-film formation composed of zinc layered hydroxides with intercalated organic molecules through the biomineralization-inspired approach utilizing amorphous states as precursors. The amorphous precursors for zinc layered hydroxides with intercalated organic molecules were prepared in the presence of poly(acrylic acid). Polymer thin-film matrices were immersed in aqueous solution containing the amorphous precursors in order to develop hybrid thin films. The structures of resultant hybrid thin films composed of layered zinc hydroxides have been examined with polarizing optical microscopy, scanning electron microscopy and X-ray diffraction . These characterizations revealed that the hybrid thin films of layered zinc hydroxides exhibit macroscopically ordered structures. Hybrid thin films comprising of the layered zinc hydroxides are converted to ZnO thin films through thermal treatment. The resultant ZnO thin films exhibit macroscopically ordered structures. It is elucidated that ordered structures of ZnO thin films depend on the molecule structures of intercalated guest molecules as well as the original structures of hybrid thin films of layered zinc hydroxides. These results suggest that the biomineralization-approach is useful for the development of functional ZnO materials with ordered structures.
 Bäuerlein, E.; Behrens, P.; Epple, M. Handbook of Biomineralization, Wiely-VCH, Weinheim, 2007.
 Kato, T.; Sakamoto, T.; Nishimura, T. MRS Bull. 2010, 35, 127
 Arakaki, A.; Shimizu, K.; Oda, M.; Sakamoto, T.; Nishimura, T.; Kato, T. Org. Biomol. Chem. 2015, 13, 974.
 Kajiyama, S.; Nishimura, T.; Sakamoto, T.; Kato, T. Small 2014, 10, 1634.
 Aizenberg, J.; Muller, D. A.; Grazul, J. L.; Hamann, D. R. Science 2003, 299, 1205.
 Matsumura, S.; Horiguchi, Y.; Nishimura, T.; Sakai, H.; Kato, T. Chem. Eur. J. 2016, 22, 7094.
8:00 PM - BM09.08.12
Macromolecular Assembly of DNA into Complex Nanostructures via Hybridization Chain Reaction
Laura Lanier1,Harry Bermudez1
Univ of Massachusetts-Amherst1Show Abstract
Through the use of a macromolecular self-assembly technique called hybridization chain reaction (HCR), we have created complex, well-defined nanostructures. HCR is a supramolecular polymerization of DNA that proceeds as an isothermal cascade of strand displacement reactions. Two DNA monomers are kinetically trapped in hairpins until the addition of an initiator strand opens the hairpin of one monomer through a strand displacement reaction. The unhybridized end then opens the hairpin of the other monomer through a strand displacement reaction. This cascade of strand displacement reactions continues, producing a supramolecular DNA polymer. This project aims to demonstrate the living mechanism of HCR. Further, the living nature of HCR is used to create well-defined nanostructures of DNA by HCR in order to expand the design toolbox of DNA for potential applications in such fields as nanomedicine, sensing, synthetic biology.
We have demonstrated that HCR produces supramolecular polymers of DNA in a controlled manner through a living polymerization mechanism. Through macromolecular assembly by HCR, DNA polymers of narrow dispersity are produced whose molecular weight is controlled by the monomer to initiator stoichiometric ratio, consistent with a living polymerization mechanism. Additionally, HCR polymerization can be continued by the addition of further monomer, demonstrating its living nature by the absence of termination and chain transfer reactions. Identification of the living character of HCR presents new opportunities in macromolecular assembly of structural DNA nanotechnology and molecular biology.
Utilizing the demonstrated living nature, complex, well-defined nanostructures are created via HCR. Bottlebrush structures are created by modifying the monomer to include an additional overhang that initiates a secondary HCR polymerization with aspect ratio controlled by the stoichiometric ratios of the initiator and monomer strands of each HCR sequence. Supramolecular star polymers are created by modifying a four-arm star to initiate HCR from each arm. The growth of each arm is independently controlled, allowing for the creation of asymmetric DNA star polymers. The creation of these complex nanostructures is demonstrated by gel electrophoresis and atomic force microscopy.
8:00 PM - BM09.08.13
Enzymatically Activated Aggregation and Cell-Adhesion of Peptide-Nanoparticle Conjugates for Surface-Enhanced Raman Spectroscopy (SERS) Based Diagnostics and Imaging
Hailin Huang1,2,3,Stephen O'Brien1,3,Duncan Graham4,Rein Ulijn2,5,3
The City College of New York1,CUNY Advance Science Research Center2,The Graduate Center, City University of New York3,University of Strathclyde4,Hunter College5Show Abstract
In cancer research, multi-functionalized gold nanoparticles (GNPs) have advanced to realize simultaneous diagnosis and therapy (i.e. theranostics) due to their excellent optical properties and accessibility of surface modification. An example of GNPs used for cancer theranostics is to functionalize the surface using Raman reporter molecules for surface enhanced Raman spectroscopy (SERS) and incorporate anti-cancer drug molecules on the surface. One critical aspect of using nanomedicines for cancer treatment is to assure that the administered drugs reach and accumulate at the tumor site, rather than retaining at healthy parts of the body or being eliminated by the reticuloendothelial system. Unfortunately, during the past 10 years, the delivery efficiency of nanoparticles has not been improved significantly, and only a median of 0.7% of injected dose of nanoparticles reached the tumor. Increasing the delivery efficiency is therefore one of the greatest challenges for the development of cancer nanomedicines.
In this study, an enzyme responsive peptide functionalized GNP is designed to target metastatic tumor cells with a new dual-action targeting mechanism. Immobilized peptides with which contain an enzyme-cleavable linker which incorporates a cryptic adhesive ligand are immobilized onto GNP and recognized and cleaved by collagenase MMP-9. MMP-9 is overexpressed by malignant tumor cells and plays a central role in metastatic cancer progression by degrading proteins in the extracellular matrix (ECM). The enzymatic product fragments retained on the surface of GNP, LRGDC, trigger nanoparticle self-assembly and enhances SERS signals. Moreover, the RGD motif binds preferentially to αvβ3 integrins which are protein receptors overexpressed on tumoral endothelial cells primarily during angiogenesis. The advantage of this design is that the surface functionalized GNPs remain an inactive state through the blood circulation and become active once they reach the tumor ECM, not only exposing the RGD surface for cell binding, but also enabling tumor diagnosis by SERS. This active targeting mechanism can potentially reduce the binding of RGD to non-tumoral integrins and hence increase the delivery efficiency of the nanoparticles.
1. Zhong, J.; Cobb, S. L.; Cameron, N. R. Biomater. Sci. 2017, 5 (5), 872.
2. Laing, S.; Jamieson, L. E.; Faulds, K.; Graham, D. Nat Rev Chem. 2017, 1, 1.
3. Wilhelm, S.; Tavares, A. J.; Dai, Q. et al. Nat. Rev. Mater. 2016, 1 (5), 16014.
4. Kalafatovic, D.; Nobis, M.; Son, J.; Anderson, K. I.; Ulijn, R. V. Biomaterials. 2016, 98, 192.
5. Roberts, J.; Sahoo, J.; Ulijn, R. V. et al. ACS Nano. 2016, 10 (7), 6667.
6. Sahoo, J.; Graham, D.; Ulijn, R. V. et al. Chem. Comm. 2016, 52 (25), 4698.
8:00 PM - BM09.08.14
Octopus-Inspired Adhesive and Conductive Patch Sensor for Biosignal Monitoring
SeungHoon Choi1,HeonJoon Lee2,Changhyun Pang1,2,3
Sungkyunkwan University Advanced Institute of NanoTechnology1,Sungkyunkwan University2,Samsung Advanced Institute for Health Science and Technology3Show Abstract
The attachment phenomena of various hierarchical architectures found in nature have extensively drawn attention for developing highly biocompatible adhesives for skin or wet inner organs. Scientists have reported bioinspired skin adhesives with various multiscale architectures including patches with mushroom-shaped tips with or without conductive materials, microneedles, and miniaturized octopus-like suction cups. Adhesives with mushroom-shaped tips have demonstrated enhanced attachment by van der Waals interactions on dry skin, and stabilized contact to active skin surface underwater by embedding the patch underneath swimwear. These adhesives, however, cannot maintain adequate adhesion on wet skin or skin under flowing water. Microneedle patches have indicated striking adhesion performances through mechanical interlocking, but they are more appropriate for wound closure or invasive therapies rather than reversible and residue-free dermal attachment. In recent years, hierarchical structures of octopus suckers have been investigated for their unique reversible adhesion in both dry and wet conditions. The octopus sucker can be divided into two parts: 1) the protruded cup-like upper portion (infundibulum) and 2) the lower portion with a dome-like protuberance (acetabulum). For such adhesive capabilities, suction cups of octopi have been mimicked to develop reusable and residual-free skin patches for versatile medical applications. Here, we present an octopus-inspired skin-adhesive with meniscus-controlled unfoldable 3D microsuckers in micropillars, as well as its application to a stretchable patch sensor composed of carbon-based conducting polymer composite (CPC) films. Mimicking the rim and infundibulum of octopus suction cups, the microsuckers are fabricated by controlling the wetting properties of a liquid precursor during simple molding. Moreover, the adhesive shows strong dry/wet adhesion performances in both pull-off and peeling-off directions against a wafer and rough, hairy skin. Finally, the patch sensor incorporated with 3D microsuckers displays sensitive and reliable piezoresistive responses to lateral strain and vertical pressure. With high conformity on human skin and water-resistant nature, our patch sensor demonstrates efficient detection of not only electrocardiogram (ECG), but also the motion of a human finger even in an underwater environment. We believe that this work represents a timely, methodological advance in nature and breakthrough in the fields of wearable and skin-attachable sensor devices for future healthcare applications.
8:00 PM - BM09.08.15
Antimicrobial Modification of K-Wires via Novel Polymer Grafting Technology
Mikhail Bredikhin1,Dmitry Gil2,Christopher Gross3,Igor Luzinov1,Alexey Vertegel1
Clemson University1,Harvard University2,Medical University of South Carolina3Show Abstract
Introduction: Kirschner wires are the external smooth stainless-steel pins that are used today in bone fracture fixation. These wires provide the surface for bacteria to adhere onto and form a biofilm, which makes them considerably less-susceptible to antibiotics. The solution to this problem has been an active area of research.
Surface coatings of implants with bioactive molecules is a modern approach to modify implant’s function. However, in the case of K-wires, coating the pin with an antibiotic would not have much effect in vivo since this coating is poorly adherent and would be easily removed when the wire is drilled by a surgeon. In this work, we propose a novel method of grafting the K-wires with highly adherent polymer. Specifically, this study utilizes cross-linkable random "brush" copolymer of OEGMA, GMA, and LMA, which can be covalently attached to solid surfaces. Such polymeric coating can be loaded with an antibiotic, allowing both the protection of the antimicrobial coating during insertion into the bone and optimal release over time.
Materials&Methods: Gentamicin sulfate (GS), butanone-2 (MEK) and other materials were purchased from Sigma-Aldrich. The polymeric “brush" is synthesized via solution polymerization. Grafting of the polymer onto the surface was performed by dip-coating. The wires were dipped in the MEK solution of the polymer. Following thermal cross-linking of the polymer, the polymer-coated wires were submerged and kept in aqueous GS solution for 24 hours. Finally, the wires