Tropoelastin modulates systemic and local tissue responses to enhance wound healing.

Wound healing is facilitated by biomaterials-based grafts and substantially impacted by orchestrated inflammatory responses that are essential to the normal repair process. Tropoelastin (TE) based materials are known to shorten the period for wound repair but the mechanism of anti-inflammatory performance is not known. To explore this, we compared the performance of the gold standard Integra Dermal Regeneration Template (Integra), polyglycerol sebacate (PGS), and TE blended with PGS, in a murine full-thickness cutaneous wound healing study. Systemically, blending with TE favorably increased the F4/80+ macrophage population by day 7 in the spleen and contemporaneously induced elevated plasma levels of anti-inflammatory IL-10. In contrast, the PGS graft without TE prompted prolonged inflammation, as evidenced by splenomegaly and greater splenic granulocyte and monocyte fractions at day 14. Locally, the inclusion of TE in the graft led to increased anti-inflammatory M2 macrophages and CD4+T cells at the wound site, and a rise in Foxp3+ regulatory T cells in the wound bed by day 7. We conclude that the TE-incorporated skin graft delivers a pro-healing environment by modulating systemic and local tissue responses. STATEMENT OF SIGNIFICANCE: Tropoelastin (TE) has shown significant benefits in promoting the repair and regeneration of damaged human tissues. In this study, we show that TE promotes an anti-inflammatory environment that facilitates cutaneous wound healing. In a mouse model, we find that inserting a TE-containing material into a full-thickness wound results in defined, pro-healing local and systemic tissue responses. These findings advance our understanding of TE's restorative value in tissue engineering and regenerative medicine, and pave the way for clinical applications.

Decohesion of a polyolefin overlay from a substrate high density polyethylene layer by impact induced stress waves.

High-density polythene (HDPE) is difficult to separate from food packaging waste for recycling because the packaging occasionally has multilayer plastic labels attached. Solvents are employed in the current separation techniques to remove undesirable layers from HDPE substrates. The possibility of separating HDPE via the impact-delamination phenomenon was explored both theoretically and experimentally. Using the cohesive zone model (CZM), the decohesion of layers in a model two-layer laminate made of HDPE and LDPE layers was studied theoretically. According to this study, stress waves emerge and severely damage the adhesion between the layers as a cutting blade strikes the laminate at speeds greater than 40 m/s. The damage can be enhanced by increasing the strike velocity and the apex radius of the blade. These findings show that a novel plastic delaminator that can cut and delaminate the laminates simultaneously can be designed. The proposed machine will feature two sets of blades with varying edge apex radii. One set of blades can be designed to cause the most adhesion damage while the other blades cut the laminate. This unique combination of cutting and delamination operations has several benefits, including less solvent waste and downstream processes, greater environmental friendliness, and faster HDPE separation. Laminates from HDPE milk bottles were cut using a high-speed cutter-blender with six blades to test the predicted results. The cut HDPE flakes were separated pneumatically. According to FTIR analysis and SEM, only a trace of adhesive was present on the cut and separated HDPE flakes.

Rapid Regeneration of a Neoartery with Elastic Lamellae.

Native arteries contain a distinctive intima-media composed of organized elastin and an adventitia containing mature collagen fibrils. In contrast, implanted biodegradable small-diameter vascular grafts do not present spatially regenerated, organized elastin. The elastin-containing structures within the intima-media region encompass the elastic lamellae (EL) and internal elastic lamina (IEL) and are crucial for normal arterial function. Here, the development of a novel electrospun small-diameter vascular graft that facilitates de novo formation of a structurally appropriate elastin-containing intima-media region following implantation is described. The graft comprises a non-porous microstructure characterized by tropoelastin fibers that are embedded in a PGS matrix. After implantation in mouse abdominal aorta, the graft develops distinct cell and extracellular matrix profiles that approximate the native adventitia and intima-media by 8 weeks. Within the newly formed intima-media region there are circumferentially aligned smooth muscle cell layers that alternate with multiple EL similar to that found in the arterial wall. By 8 months, the developed adventitia region contains mature collagen fibrils and the neoartery presents a distinct IEL with thickness comparable to that in mouse abdominal aorta. It is proposed that this new class of material can generate the critically required, organized elastin needed for arterial regeneration.

Aqueous Polymeric Hollow Particles as an Opacifier by Emulsion Polymerization Using Macro-RAFT Amphiphiles.

A robust polymerization technique that enables the surfactant-free aqueous synthesis of a high solid content latex containing polymeric hollow particles is presented. Uniquely designed amphiphilic macro-reversible addition fragmentation chain transfer (RAFT) copolymers were used as sole stabilizers for monomer emulsification as well as for free-radical emulsion polymerization. The polymerization was found to be under RAFT control, generating various morphologies from spherical particles, wormlike structures to polymer vesicles. The final particles were dominantly polymeric vesicles which had a substantially uniform and continuous polymer layer around a single aqueous filled void. They produced hollow particles once dried and were successfully used as opacifiers to impart opacity into polymer paint films. This method is simple, can be performed in a controllable and reproducible manner, and may be performed using diverse procedures.

Steric Stabilization of γ-Fe2O3 Superparamagnetic Nanoparticles in a Hydrophobic Ionic Liquid and the Magnetorheological Behavior of the Ferrofluid.

Hydrophobic ionic liquid ferrofluids (ILFFs) are studied for use in electrospray thrusters for microsatellite propulsion under nonatmospheric and in high-temperature environments. We synthesized a hydrophobic ILFF by dispersing sterically stabilized γ-Fe2O3 nanoparticles (NPs) in the ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide. A diblock copolymer, C4-RAFT-AA10-DEAm60, was synthesized to facilitate multipoint bidentate anchoring to the NP through the acrylic acid block. The DEAm60 layer was incorporated to generate steric repulsion between particles to protect against the aggregation of magnetized particles arising from dipole-dipole attraction. The effect of shearing and variation in the magnetic field strength on the steric repulsion was examined using the DLVO theory. The effect of varying the magnetic field strength and particle concentration on the viscoelastic properties of the ferrofluid was evaluated using rheometry. The viscosity of the ferrofluid increased with the magnetic field strength, indicating that the magnetized particles assembled into a structure. The level of straining required to break down the structure formed by the magnetized particles increased with the magnetic field strength and particle concentration. The absence of particle interlocking during shearing was indicated by the smooth viscosity versus shear rate traces. The DLVO analysis showed that increasing the magnetic attraction between the particles causes the DEAm60 brush layers on the particles to overlap more, resulting in an increase in the steric repulsion. As overlapping increases, osmotic repulsion is caused before progressing to a strong elastic repulsion. The effect of the polymer solubility and particle interaction due to hydrodynamic forces on the steric repulsion was also analyzed.

Biodistribution and Clearance of Stable Superparamagnetic Maghemite Iron Oxide Nanoparticles in Mice Following Intraperitoneal Administration.

Nanomedicine is an emerging field with great potential in disease theranostics. We generated sterically stabilized superparamagnetic iron oxide nanoparticles (s-SPIONs) with average core diameters of 10 and 25 nm and determined the in vivo biodistribution and clearance profiles. Healthy nude mice underwent an intraperitoneal injection of these s-SPIONs at a dose of 90 mg Fe/kg body weight. Tissue iron biodistribution was monitored by atomic absorption spectroscopy and Prussian blue staining. Histopathological examination was performed to assess tissue toxicity. The 10 nm s-SPIONs resulted in higher tissue-iron levels, whereas the 25 nm s-SPIONs peaked earlier and cleared faster. Increased iron levels were detected in all organs and body fluids tested except for the brain, with notable increases in the liver, spleen, and the omentum. The tissue-iron returned to control or near control levels within 7 days post-injection, except in the omentum, which had the largest and most variable accumulation of s-SPIONs. No obvious tissue changes were noted although an influx of macrophages was observed in several tissues suggesting their involvement in s-SPION sequestration and clearance. These results demonstrate that the s-SPIONs do not degrade or aggregate in vivo and intraperitoneal administration is well tolerated, with a broad and transient biodistribution. In an ovarian tumor model, s-SPIONs were shown to accumulate in the tumors, highlighting their potential use as a chemotherapy delivery agent.

Tunable and noncytotoxic PET/SPECT-MRI multimodality imaging probes using colloidally stable ligand-free superparamagnetic iron oxide nanoparticles.

Physiologically stable multimodality imaging probes for positron emission tomography/single-photon emission computed tomography (PET/SPECT)-magnetic resonance imaging (MRI) were synthesized using the superparamagnetic maghemite iron oxide (γ-Fe2O3) nanoparticles (SPIONs). The SPIONs were sterically stabilized with a finely tuned mixture of diblock copolymers with either methoxypolyethylene glycol (MPEG) or primary amine NH2 end groups. The radioisotope for PET or SPECT imaging was incorporated with the SPIONs at high temperature. 57Co2+ ions with a long half-life of 270.9 days were used as a model for the radiotracer to study the kinetics of radiolabeling, characterization, and the stability of the radiolabeled SPIONs. Radioactive 67Ga3+ and Cu2+-labeled SPIONs were also produced successfully using the optimized conditions from the 57Co2+-labeling process. No free radioisotopes were detected in the aqueous phase for the radiolabeled SPIONs 1 week after dispersion in phosphate-buffered saline (PBS). All labeled SPIONs were not only well dispersed and stable under physiological conditions but also noncytotoxic in vitro. The ability to design and produce physiologically stable radiolabeled magnetic nanoparticles with a finely controlled number of functionalizable end groups on the SPIONs enables the generation of a desirable and biologically compatible multimodality PET/SPECT-MRI agent on a single T2 contrast MRI probe.

Fluorescent Labeling and Biodistribution of Latex Nanoparticles Formed by Surfactant-Free RAFT Emulsion Polymerization.

The authors report the preparation of a novel range of functional polyacrylamide stabilized polystyrene nanoparticles, obtained by surfactant-free reversible addition-fragmentation chain transfer (RAFT) emulsion polymerization, their fluorescent tagging, cellular uptake, and biodistribution. The authors show the versatility of the RAFT emulsion process for the design of functional nanoparticles of well-defined size that can be used as drug delivery vectors. Functionalization with a fluorescent tag offers a useful visualization tool for tracing, localization, and clearance studies of these carriers in biological models. The studies are carried out by labeling the sterically stabilized latex particles chemically with rhodamine B. The fluorescent particles are incubated in a healthy human renal proximal tubular cell line model, and intravenously injected into a mouse model. Cellular localization and biodistribution of these particles on the biological models are explored.

Durable Superhydrophobic Surfaces via Spontaneous Wrinkling of Teflon AF.

We report the fabrication of both single-scale and hierarchical superhydrophobic surfaces, created by exploiting the spontaneous wrinkling of a rigid Teflon AF film on two types of shrinkable plastic substrates. Sub-100 nm to micrometric wrinkles were reproducibly generated by this simple process, with remarkable control over the size and hierarchy. Hierarchical Teflon AF wrinkled surfaces showed extremely high water repellence (contact angle 172°) and very low contact angle hysteresis (2°), resulting in droplets rolling off the surface at tilt angles lower than 5°. The wrinkling process intimately binds the Teflon AF layer with its substrate, making these surfaces mechanically robust, as revealed by macroscale and nanoscale wear tests: hardness values were close to that of commercial optical lenses and aluminum films, resistance to scratch was comparable to commercial hydrophobic coatings, and damage by extensive sonication did not significantly affect water repellence. By this fabrication method the size of the wrinkles can be reproducibly tuned from the nanoscale to the microscale, across the whole surface in one step; the fabrication procedure is extremely rapid, requiring only 2 min of thermal annealing to produce the desired topography, and uses inexpensive materials. The very low roll-off angles achieved in the hierarchical surfaces offer a potentially up-scalable alternative as self-cleaning and drag-reducing coatings.

Preparation of Inert Polystyrene Latex Particles as MicroRNA Delivery Vectors by Surfactant-Free RAFT Emulsion Polymerization.

We present the preparation of 11 nm polyacrylamide-stabilized polystyrene latex particles for conjugation to a microRNA model by surfactant-free RAFT emulsion polymerization. Our synthetic strategy involved the preparation of amphiphilic polyacrylamide-block-polystyrene copolymers, which were able to self-assemble into polymeric micelles and "grow" into polystyrene latex particles. The surface of these sterically stabilized particles was postmodified with a disulfide-bearing linker for the attachment of the microRNA model, which can be released from the latex particles under reducing conditions. These nanoparticles offer the advantage of ease of preparation via a scaleable process, and the versatility of their synthesis makes them adaptable to a range of applications.

The interaction of sterically stabilized magnetic nanoparticles with fresh human red blood cells.

Sterically stabilized superparamagnetic iron oxide nanoparticles (SPIONs) were incubated with fresh human erythrocytes (red blood cells [RBCs]) to explore their potential application as magnetic resonance imaging contrast agents. The chemical shift and linewidth of (133)Cs(+) resonances from inside and outside the RBCs in (133)Cs nuclear magnetic resonance spectra were monitored as a function of time. Thus, we investigated whether SPIONs of two different core sizes and with three different types of polymeric stabilizers entered metabolically active RBCs, consuming glucose at 37°C. The SPIONs broadened the extracellular (133)Cs(+) nuclear magnetic resonance, and brought about a small change in its chemical shift to a higher frequency; while the intracellular resonance remained unchanged in both amplitude and chemical shift. This situation pertained over incubation times of up to 90 minutes. If the SPIONs had entered the RBCs, the intracellular resonance would have become broader and possibly even shifted. Therefore, we concluded that our SPIONs did not enter the RBCs. In addition, the T 2 relaxivity of the small and large particles was 368 and 953 mM(-1) s(-1), respectively (three and nine times that of the most effective commercially available samples). This suggests that these new SPIONs will provide a superior performance to any others reported thus far as magnetic resonance imaging contrast agents.

Premature detonation of an NH4NO3 emulsion in reactive ground.

When NH4NO3 emulsions are used in blast holes containing pyrite, they can exothermally react with pyrite, causing the emulsion to intensively heat and detonate prematurely. Such premature detonations can inflict fatal and very costly damages. The mechanism of heating of the emulsions is not well understood though such an understanding is essential for designing safe blasting. In this study the heating of an emulsion in model blast holes was simulated by solving the heat equation. The physical factors contributing to the heating phenomenon were studied using microscopic and calorimetric methods. Microscopic studies revealed the continuous formation of a large number of gas bubbles as the reaction progressed at the emulsion-pyrite interface, which made the reacting emulsion porous. Calculations show that the increase in porosity causes the thermal conductivity of a reacting region of an emulsion column in a blast hole to decrease exponentially. This large reduction in the thermal conductivity retards heat dissipation from the reacting region causing its temperature to rise. The rise in temperature accelerates the exothermic reaction producing more heat. Simulations predict a migration of the hottest spot of the emulsion column, which could dangerously heat the primers and boosters located in the blast hole.

Self-assembling array of magnetoelectrostatic jets from the surface of a superparamagnetic ionic liquid.

Electrospray is a versatile technology used, for example, to ionize biomolecules for mass spectrometry, create nanofibers and nanowires, and propel spacecraft in orbit. Traditionally, electrospray is achieved via microfabricated capillary needle electrodes that are used to create the fluid jets. Here we report on multiple parallel jetting instabilities realized through the application of simultaneous electric and magnetic fields to the surface of a superparamagnetic electrically conducting ionic liquid with no needle electrodes. The ionic liquid ferrofluid is synthesized by suspending magnetic nanoparticles in a room-temperature molten salt carrier liquid. Two ILFFs are reported: one based on ethylammonium nitrate (EAN) and the other based on EMIM-NTf2. The ILFFs display an electrical conductivity of 0.63 S/m and a relative magnetic permeability as high as 10. When coincident electric and magnetic fields are applied to these liquids, the result is a self-assembling array of emitters that are composed entirely of the colloidal fluid. An analysis of the magnetic surface stress induced on the ILFF shows that the electric field required for transition to spray can be reduced by as much as 4.5 × 10(7) V/m compared to purely electrostatic spray. Ferrofluid mode studies in nonuniform magnetic fields show that it is feasible to realize arrays with up to 16 emitters/mm(2).

Ultrasmall superparamagnetic iron oxide nanoparticle prelabelling of human neural precursor cells.

Stem cells prelabelled with iron oxide nanoparticles can be visualised using magnetic resonance imaging (MRI). This technique allows for noninvasive long-term monitoring of migration, integration and stem cell fate following transplantation into living animals. In order to determine biocompatibility, the present study investigated the biological impact of introducing ultrasmall superparamagnetic iron oxide nanoparticles (USPIOs) into primary human fetal neural precursor cells (hNPCs) in vitro. USPIOs with a mean diameter of 10-15 nm maghemite iron oxide core were sterically stabilised by 95% methoxy-poly(ethylene glycol) (MPEG) and either 5% cationic (NH2) end-functionalised, or 5% Rhodamine B end-functionalised, polyacrylamide. The stabilising polymer diblocks were synthesised by reversible addition-fragmentation chain transfer (RAFT) polymerisation. Upon loading, cellular viability, total iron capacity, differentiation, average distance of migration and changes in intracellular calcium ion concentration were measured to determine optimal loading conditions. Taken together we demonstrate that prelabelling of hNPCs with USPIOs has no significant detrimental effect on cell biology and that USPIOs, when utilised at an optimised dosage, are an effective means of noninvasively tracking prelabelled hNPCs.

Phase behavior of amphiphilic diblock co-oligomers with nonionic and ionic hydrophilic groups.

The synthesis of a series of co-oligomer amphiphiles by RAFT and their self-assembly behavior in water is described. These novel amphiphiles, comprised of styrene, butyl acrylate, and alkyl hydrophobes together with ionic acrylic acid and nonionic hydroxyethylacrylate hydrophilic moieties and with a total degree of polymerization from 5 to 17, represent a new class of small-molecule surfactants that can be formed from the immense potential library of all polymerizable monomers. Examples of micellar solutions and discrete cubic, hexagonal, lamellar, and inverted hexagonal lyotropic phases, as well as vesicle dispersions and coexisting lamellar phases, are reported and characterized by small-angle scattering. The variation of self-assembly structure with co-oligomer composition, concentration, and solution conditions is interpreted by analogy with the surfactant packing parameter used for conventional small-molecule amphiphiles.

The composition and end-group functionality of sterically stabilized nanoparticles enhances the effectiveness of co-administered cytotoxins.

Diffusion of active cytotoxic agents throughout an entire solid tumour is a particular challenge to successful drug delivery. Here we show the simple and robust generation of non-toxic, 10-15 nm superparamagnetic iron oxide nanoparticles (SPIONs) that have been sterically stabilized by either 100% anionic or 100% cationic or 100% neutral end-functionalized steric stabilizers or by novel combinations of cationic and neutral end-functionalized polymer. When these nanoparticles were co-administered with various anti-cancer drugs, a significant increase in the diffusion and effectiveness of the cytotoxin in a 3-dimensional model of a solid tumour was shown for specific combinations of surface functionality and cytotoxin. The critical determinant of enhanced cytotoxin diffusion and effectiveness was the end functionality of the steric stabilizers and not the core composition (either iron oxide, silica or gold). We provide evidence that SPIONs stabilized with heterogeneous steric stabilizers enhance nuclear uptake of doxorubicin across multiple cell layers.

Stable and water-tolerant ionic liquid ferrofluids.

Ionic liquid ferrofluids have been prepared containing both bare and sterically stabilized 8-12 nm diameter superparamagnetic iron oxide nanoparticles, which remain stable for several months in both protic ethylammonium and aprotic imidazolium room-temperature ionic liquids. These ferrofluids exhibit spiking in static magnetic fields similar to conventional aqueous and nonaqueous ferrofluids. Ferrofluid stability was verified by following the flocculation and settling behavior of dilute nanoparticle dispersions. Although bare nanoparticles showed excellent stability in some ILs, they were unstable in others, and exhibited limited water tolerance. Stability was achieved by incorporating a thin polymeric steric stabilization layer designed to be compatible with the IL. This confers the added benefit of imbuing the ILF with a high tolerance to water.

Optimized steric stabilization of aqueous ferrofluids and magnetic nanoparticles.

The preparation and properties of an aqueous ferrofluid consisting of a concentrated (>65 wt %) dispersion of sterically stabilized superparamagnetic, iron oxide (maghemite) nanoparticles stable for several months at high ionic strength and over a broad pH range is described. The 6-8 nm diameter nanoparticles are individually coated with a short poly(acrylic acid)-b-poly(acrylamide) copolymer, designed to form the thinnest possible steric stabilizing layer while remaining strongly attached to the iron oxide surface over a wide range of nanoparticle concentrations. Thermogravimetric analysis yields an iron oxide content of 76 wt % in the dried particles, consistent with a dry polymer coating of approximately 1 nm in thickness, while the poly(acrylamide) chain length indicated by electrospray mass spectrometry is consistent with the 4-5 nm increase in the hydrodynamic radius observed by light scattering when the poly(acrylamide) stabilizing chains are solvated. Saturation magnetization experiments indicate nonmagnetic surface layers resulting from the strong chemical attachment of the poly(acrylic acid) block to the particle surface, also observed by Fourier transform infrared spectroscopy.

Controlling the locus of bubble nucleation by dissolved gases in heterogeneous liquid-liquid systems.

We have examined the nucleation of chemically generated nitrogen gas bubbles in microheterogeneous systems, using optical microscopy on a model system consisting of a single liquid-liquid interface. Results clearly show that bubble nucleation occurs in both the aqueous and oil phases, despite the nitrogen production reaction being a purely aqueous phase process. A theoretical model is developed which describes the time evolution of the nitrogen concentration profile, and this reveals that bubbles in the oil are a result of homogeneous nucleation of dissolved N(2) transported across the interface into a (supersaturated) diffusion layer. We further show that bubble nucleation in the oil can be inhibited or eliminated by adding water-soluble surfactants, which facilitates aqueous phase bubble nucleation and then acts as highly effective nitrogen sinks, severely reducing the flux of dissolved gas across the water-oil interface.