Lilo D. Pozzo

A synthetic fibrin cross-linking polymer for modulating clot properties and inducing hemostasis

By cross-linking fibrin, intravenously injected factor XIII–inspired hemostatic polymers stabilize clots and reduce bleeding in a rat model of trauma and fluid resuscitation. Bioinspired polymers fortify blood clots Major blood loss is a leading cause of death after trauma, and currently available intravenously delivered clotting agents are expensive, require special storage, have limited shelf life, and carry risk of immunogenicity. In this issue, Chan et al. describe an off-the-shelf, synthetic, bioinspired polymer called PolySTAT that stops the bleeding after trauma and restores hemostasis. In healthy animals, the polymer flows throughout the body, harmless and passive. Upon encountering a blood clot at a site of vascular injury, PolySTAT cross-links the fledgling fibrin matrix much like the transglutaminase factor XIII, strengthening the clot and fortifying it against degrading enzymes that are overactive in trauma. In a rat model of femoral artery injury and fluid resuscitation, PolySTAT or control therapies were injected immediately after the onset of bleeding. All animals treated with PolySTAT survived, whereas only 4 of the remaining 20 animals in various control groups survived. Additionally, none of the PolySTAT-treated animals experienced rebleeding, suggesting that the clots formed were strong, despite increased blood pressure from the infusions. This polymer-based solution to clotting could be a welcomed addition in critical care medicine, because its specificity minimizes risk of distant thrombosis (for example, in the heart or lungs) and has scalable manufacturing properties as well as favorable biodistribution and pharmacokinetics. Further preclinical studies will be needed in larger animal models to evaluate not only clotting function but also the extent of organ injury in surviving animals. Clotting factor replacement is the standard management of acute bleeding in congenital and acquired bleeding disorders. We present a synthetic approach to hemostasis using an engineered hemostatic polymer (PolySTAT) that circulates innocuously in the blood, identifies sites of vascular injury, and promotes clot formation to stop bleeding. PolySTAT induces hemostasis by cross-linking the fibrin matrix within clots, mimicking the function of the transglutaminase factor XIII. Furthermore, synthetic PolySTAT binds specifically to fibrin monomers and is uniformly integrated into fibrin fibers during fibrin polymerization, resulting in a fortified, hybrid polymer network with enhanced resistance to enzymatic degradation. In vivo hemostatic activity was confirmed in a rat model of trauma and fluid resuscitation in which intravenous administration of PolySTAT improved survival by reducing blood loss and resuscitation fluid requirements. PolySTAT-induced fibrin cross-linking is a novel approach to hemostasis using synthetic polymers for noninvasive modulation of clot architecture with potentially wide-ranging therapeutic applications.

Sono-photoacoustic imaging of gold nanoemulsions: Part I. Exposure thresholds

Integrating high contrast bubbles from ultrasound imaging with plasmonic absorbers from photoacoustic imaging is investigated. Nanoemulsion beads coated with gold nanopsheres (NEB-GNS) are excited with simultaneous light (transient heat at the GNSs) and ultrasound (rarefactional pressure) resulting in a phase transition achievable under different scenarios, enhancing laser-induced acoustic signals and enabling specific detection of nanoprobes at lower concentration. An automated platform allowed dual parameter scans of both pressure and laser fluence while recording broadband acoustic signals. Two types of NEB-GNS and individual GNS were investigated and showed the great potential of this technique to enhance photoacoustic/acoustic signals. The NEB-GNS size distribution influences vaporization thresholds which can be reached at both permissible ultrasound and light exposures at deep penetration and at low concentrations of targets. This technique, called sono-photoacoustics, has great potential for targeted molecular imaging and therapy using compact nanoprobes with potentially high-penetrability into tissue.

Laser-induced cavitation in nanoemulsion with gold nanospheres for blood clot disruption: in vitro results

Optically activated cavitation in a nanoemulsion contrast agent is proposed for therapeutic applications. With a 56°C boiling point perfluorohexane core and highly absorptive gold nanospheres at the oil-water interface, cavitation nuclei in the core can be efficiently induced with a laser fluence below medical safety limits (70 mJ/cm2 at 1064 nm). This agent is also sensitive to ultrasound (US) exposure and can induce inertial cavitation at a pressure within the medical diagnostic range. Images from a high-speed camera demonstrate bubble formation in these nanoemulsions. The potential of using this contrast agent for blood clot disruption is demonstrated in an in vitro study. The possibility of simultaneous laser and US excitation to reduce the cavitation threshold for therapeutic applications is also discussed.

Correlating Structure and Photocurrent for Composite Semiconducting Nanoparticles with Contrast Variation Small-Angle Neutron Scattering and Photoconductive Atomic Force Microscopy

Aqueous dispersions of semiconducting nanoparticles have shown promise as a robust and scalable platform for the production of efficient polymer/fullerene active layers in organic photovoltaic applications. Semiconducting nanoparticles are a composite of both an n-type and p-type semiconductor contained within a single nanoparticle. In order to realize efficient organic solar cells from these materials, there is a need to understand how the size and internal distribution of materials within each nanoparticle contributes to photocurrent generation in a nanoparticle-derived device. Therefore, characterizing the internal distribution of conjugated polymer and fullerene within the dispersion is the first step to improving performance. To date, study of polymer/fullerene structure within these nanoparticles has been limited to microscopy techniques of deposited nanoparticles. In this work, we use contrast variation with small-angle neutron scattering to determine the internal distribution of poly(3-hexylthiophene) and [6,6]phenyl-C61-butyric acid methyl ester inside the composite nanoparticles as a function of formulation while in dispersion. On the basis of these measurements, we connect the formulation of these nanoparticles with their internal structure. Using electrostatic deposited monolayers of these nanoparticles, we characterize intrinsic charge generation using photoconductive atomic force microscopy and correlate this with structures determined from small-angle neutron scattering measurements. These techniques combined show that the best performing composite nanoparticles are those that have a uniform distribution of conjugated polymer and fullerene throughout the nanoparticle volume such that electrons and holes are easily transported out of the particle.

Designing Two-Dimensional Protein Arrays through Fusion of Multimers and Interface Mutations

We have combined fusion of oligomers with cyclic symmetry and alanine substitutions to eliminate clashes and produce proteins that self-assemble into 2-D arrays upon addition of calcium ions. Using TEM, AFM, small-angle X-ray scattering, and fluorescence microscopy, we show that the designed lattices which are 5 nm high with p3 space group symmetry and 7.25 nm periodicity self-assemble into structures that can exceed 100 μm in characteristic length. The versatile strategy, experimental approach, and hexagonal arrays described herein should prove valuable for the engineering of functional nanostructured materials in 2-D.

Solvatochromism and Conformational Changes in Fully Dissolved Poly(3-alkylthiophene)s

Absorption spectroscopy is commonly utilized to probe optical properties that can be related, among other things, to the conformation of single, isolated conjugated polymer chains in solution. It is frequently suggested that changes in peak positions of optical spectra result from variations in the stiffness of polymer chains in solution because this modifies the conjugation length. In this work we utilize ultraviolet-visible (UV-vis) spectroscopy, small angle neutron scattering (SANS), and all atom molecular dynamic (AA-MD) simulations to closely probe the relationship between the conformation of single-chains of poly(3-alkylthiophene)s (P3ATs) and their optical properties. SANS results show variations in the radius of gyration and Kuhn length as a function of alkyl chain length, and structure, as well as the solvent environment. Furthermore, both SANS and MD simulations show that dissolved P3HT chains are more rigid in solvents where self-assembly and crystallization are possible. Shifts in P3AT optical properties were also observed for different solvent environments. However, these changes were not correlated to the changes in polymer conformation. Furthermore, changes in optical properties could not be perfectly described by generalized solvent-solute interactions. AA-MD simulations provide new insights into specific polymer-solvent interactions not accounted for in generalized solvatochromic theory. This work highlights the need for experiments and molecular simulations that further inform the specific role of solvent molecules on local polymer conformation and on optical properties.

A conductive liquid crystal via facile doping of an n-type benzodifurandione derivative

Two n-type molecular organic semiconductors (TI-BDF1 and TI-BDF2) consisting of thiophene-substituted indolin-2-one (TI) and benzodifurandione (BDF) with different branched side-chains have been synthesized to study the effect of molecular structure on molecular order, liquid crystal (LC) properties, and charge-transport. By tuning the branching point of the side-chains, TI-BDF2 shows a preferable edge-on π-face orientation and a high degree of liquid crystallinity, resulting in 4 orders of magnitude higher electron mobility than that of TI-BDF1. Subsequent n-doping of TI-BDF2 thin film with a thermally stable phosphonium salt affords a high electrical conductivity of 0.4 S cm−1.

Polypyrrole Coated Perfluorocarbon Nanoemulsions as a Sono-Photoacoustic Contrast Agent

A new contrast agent for combined photoacoustic and ultrasound imaging is presented. It has a liquid perfluorocarbon (PFC) core of about 250 nm diameter coated by a 30 nm thin polypyrrole (PPy) doped polymer shell emulsion that represents a broadband absorber covering the visible and near-infrared ranges (peak optical extinction at 1050 nm). When exposed to a sufficiently high intensity optical or acoustic pulse, the droplets vaporize to form microbubbles providing a strong increase in imaging sensitivity and specificity. The threshold for contrast agent activation can further drastically be reduced by up to 2 orders of magnitude if simultaneously exposing them with optical and acoustic pulses. The selection of PFC core liquids with low boiling points (i.e., perfluorohexane (56 °C), perfluoropentane (29 °C), and perfluorobutane (-2 °C)) facilitates activation and reduces the activation threshold of PPy-coated emulsion contrast agents to levels well within clinical safety limits (as low as 0.2 MPa at 1 mJ/cm2). Finally, the potential use of these nanoemulsions as a contrast agent is demonstrated in a series of phantom imaging studies.

Clusters and inverse emulsions from nanoparticle surfactants in organic solvents.

A method is presented for the synthesis of self-assembling nanoparticle surfactants in nonpolar organic solvents. The method relies on the control of long-range steric repulsion imparted by grafted polystyrene and short-range attraction from short-chain thiol molecules with an alcohol or carboxylic functionality. Similar to water-based nanoparticle surfactants, these oil-dispersed materials are found to cluster in dispersion and also to stabilize oil-water interfaces to form water-in-oil emulsions. The clustering process is characterized with dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), UV-vis spectroscopy, and transmission electron microscopy (TEM). Thermogravimetric analysis (TGA) is used to quantify the surface concentration of grafted polymer, which is found to be a parameter of critical importance for the formation of stable clusters. The clustering kinetics and dispersion stability are both affected by the polymer molecular weight, surface concentration, and chemical structure of the thiol molecules that induce particle attraction. Nanometer-sized water-in-oil emulsions are formed by sonication in the presence of nanoparticle surfactants. A large broadening of the optical absorption spectrum in the NIR region is observed because of changes in the collective surface plasmon resonance of the gold particle shell.

Macroscopically aligned nanowire arrays of π-conjugated polymers via shear-enhanced crystallization

The nanoscale structure and macroscopic morphology of π-conjugated polymers are very important for their electronic application. While ordered single crystals of small molecules have been obtained via solution deposition, macroscopically aligned films of π-conjugated polymers deposited directly from solution have always required surface modification or complex pre-deposition processing of the solution. Here, ordered nanowires were obtained via shear-enhanced crystallization of π-conjugated polymers at the air–liquid–solid interface using simple deposition of the polymer solution onto an inclined substrate. The formation of macroscopically aligned nanowire arrays was found to be due to the synergy between intrinsic (π-conjugated backbone) and external (crystallization conditions) effects. The oriented nanowires showed remarkable improvement in the charge carrier mobility compared to spin-coated films as characterized in organic field-effect transistors (OFETs). Considering the simplicity and large-scale applicability, shear-enhanced crystallization of π-conjugated polymers provides a promising strategy to achieve high-performance polymer semiconductor films for electronics applications.