Regenerated keratin proteins as potential biomaterial for drug delivery

This work aims to preliminarily evaluate the reliability of regenerated keratins (RKs) in the design of microparticulate drug delivery systems by studying their processability and cytotoxicity. RKs were extracted by sulfitolysis from wool waste. A 4.5% w/w RK solution was spray‐dried, and microparticles were sterilized by steam vapor under pressure. Scanning electron microscope, sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, and Fourier transform infrared spectroscopy were used to characterize RKs and microparticles thereof. The in vitro cytotoxicity was determined by assessing the release of lactate dehydrogenase in the human monocytic cell line Tamm–Horsfall glycoprotein‐1. RK‐based microparticles with a narrow and unimodal particle size distribution (~6 µm) were obtained. They had a raisin‐like structure with a smooth surface. Both microparticle morphology and RK molecular weight were well‐preserved after sterilization. The curve fitting of the amide I bands showed that RK in the microparticles was prevalently present in the disordered/α‐helix secondary structures which made the protein soluble in water. To promote crystallization in the β‐sheet secondary structure and, therefore, water insolubility, RK‐based microparticles were immersed in an aqueous solution of acetic acid at pH 3.5 overnight. RK did not induce any appreciable cellular cytotoxicity at any of the concentrations (from 1 up to 1000 µg sterile microparticles in 1 ml cell culture medium) or time‐points (24–72 h) tested. These preliminary data suggest the feasibility of producing RK biocompatible microparticles using waste wool as starting material. Copyright © 2013 John Wiley & Sons, Ltd.     

Mesoporous Alumina from Colloidal Biotemplating of Al Clusters

A simple and green synthesis route was disclosed for the achievement of mesoporous alumina microparticles employing polysaccharide nanoparticles (α‐chitin nanorods) as templates. Pore textures can be tuned by the cationic alumina precursor. Compared to small cations, the use of Al₁₃ and Al₃₀ oxo‐hydroxo clusters leads to better defined and elongated mesopores. Electron microscopy and spectroscopic (¹³C, ²⁷Al NMR, XPS) measurements demonstrated that this is related to the effective coating of α‐chitin nanorods by these pre‐condensed colloids.     

In situ biomimetic formation of nano‐hydroxyapatite crystals on chitosan microspheres

Chitosan (CS) is a biocompatible, noncytotoxic biomaterial used before as base material for composites. On the other hand, nano‐hydroxyapatite (nHA) is one of the main components of human bones, highly used for biomedical applications. In this work, CS microspheres were produced under a W/O emulsion system. CS microspheres with calcium ions were then exposed to Na₃PO₄ solution. In situ biomimetic nHA crystals were formed on CS microspheres to generate 15.14 ± 3.15‐μm composite microspheres. The microspheres were subsequently seeded with MG63 osteoblasts to observe their cell responses. All microspheres were characterized via scanning electron microscopy (SEM), phase‐contrast photomicroscopy, and X‐ray diffraction (XRD) analysis. The results showed flake‐like shape and islet‐like growth of nHA depositions presented on the surface of the CS microspheres. In vitro tests indicated that the CS/nHA microparticles were not only biocompatible but also enhanced cell adhesion and elongation due to the in situ biomimetic synthesis method.     

Self‐Orienting Hydrogel Micro‐Buckets as Novel Cell Carriers

Hydrogel microparticles are important in materials engineering, but their applications remain limited owing to the difficulties associated with their manipulation. Herein, we report the self‐orientation of crescent‐shaped hydrogel microparticles and elucidate its mechanism. Additionally, the microparticles were used, for the first time, as micro‐buckets to carry living cells. In aqueous solution, the microparticles spontaneously rotated to a preferred orientation with the cavity facing up. We developed a geometric model that explains the self‐orienting behavior of crescent‐shaped particles by minimizing the potential energy of this specific morphology. Finally, we selectively modified the particles’ cavities with RGD peptide and exploited their preferred orientation to load them with living cells. Cells could adhere, proliferate, and be transported and released in vitro. These micro‐buckets hold a great potential for applications in smart materials, cell therapy, and biological engineering.     

pH‐responsive starch microparticles for a tumor‐targeting implant

Much progress has been made toward stimuli‐responsive polysaccharide‐based selective tumor therapy not only because polysaccharides have nontoxic biodegradability and biocompatibility but also because their stimuli‐sensitive characteristics enable the proper transport of payloads into tumors. Here, we attempted to deliver an antitumor drug, doxorubicin (DOX), using starch‐based microparticles coupled with pH‐responsive 3‐(diethylamino)propylamine. The microparticles of starch conjugated with 3‐(diethylamino)propylamine (SDEAP) allowed for the change in hydrophobicity of SDEAPs in a pH‐dependent manner. The results revealed that SDEAPs effectively carried and released DOX and selectively killed tumor cells under acidic condition. Overall, this study suggests that DOX‐loaded SDEAPs can be further explored as a strategy for applications to acidic tumor‐targeting implants owing to the drug‐deliver efficiency and tumor selectivity.     

Preparation of 5‐Fluorouracil‐Poly(L‐lactide) Microparticles Using Solution‐Enhanced Dispersion by Supercritical CO2

Summary: 5‐Fluorouracil‐poly(L ‐lactide) (5‐Fu‐PLLA) microparticles have been prepared by an SEDS process. First, the 5‐Fu is successfully micronized and is then used to produce the 5‐Fu‐PLLA microparticles. The 5‐Fu‐PLLA microparticles synthesized by the SEDS process exhibit a rather spherical shape and a narrow particle size distribution, where it ranges from 615 to 1 990 nm, with a mean particle size of 980 nm. The dichloromethane residue in the 5‐Fu‐PLLA microparticles without any further treatment is 46 ppm. The average drug load and encapsulation efficiency of the 5‐Fu‐PLLA microparticles are 3.05 and 17.8%, respectively. The rate of drug release from the 5‐Fu‐PLLA microparticles shows mainly first‐order kinetics.

Scanning electron spectroscopy image of 5‐Fu‐PLLA microparticles.     

Fibrous Networks with Incorporated Macrocycles: A Chiral Stimuli‐Responsive Supramolecular Supergelator and Its Application to Biocatalysis in Organic Media

A new and versatile, crown ether appended, chiral supergelator has been designed and synthesized based on the bis‐urea motif. The introduction of a stereogenic center improved its gelation ability significantly relative to its achiral analogue. This low‐molecular‐weight gelator forms supramolecular gels in a variety of organic solvents. It is sensitive to multiple chemical stimuli and the sol–gel phase transitions can be reversibly triggered by host–guest interactions. The gel can be used to trap enzymes and release them on demand by chemical stimuli. It stabilizes the microparticles in Pickering emulsions so that enzyme‐catalyzed organic reactions can take place in the polar phase inside the microparticles, the organic reactants diffusing through the biphasic interface from the surrounding organic phase. Because of the higher interface area between the organic and polar phases, enzyme activity is enhanced in comparison with simple biphasic systems.     

Chemically Orthogonal Three‐Patch Microparticles

Compared to two‐dimensional substrates, only a few methodologies exist for the spatially controlled decoration of three‐dimensional objects, such as microparticles. Combining electrohydrodynamic co‐jetting with synthetic polymer chemistry, we were able to create two‐ and three‐patch microparticles displaying chemically orthogonal anchor groups on three distinct surface patches of the same particle. This approach takes advantage of a combination of novel chemically orthogonal polylactide‐based polymers and their processing by electrohydrodynamic co‐jetting to yield unprecedented multifunctional microparticles. Several micropatterned particles were fabricated displaying orthogonal click functionalities. Specifically, we demonstrate novel two‐ and three‐patch particles. Multi‐patch particles are highly sought after for their potential to present multiple distinct ligands in a directional manner. This work clearly establishes a viable route towards orthogonal reaction strategies on multivalent micropatterned particles.     

Electrocatalytic Oxidation of Methanol at Lower Potentials on Glassy Carbon Electrode Modified by Platinum and Platinum Alloys Incorporated in Poly(o‐Aminophenol) Film

The electrocatalytic oxidation of methanol at a glassy carbon electrode modified by a thin film of poly(o‐aminophenol) (PoAP) containing Pt, Pt‐Ru and Pt‐Sn microparticles has been investigated using cyclic voltammetry as analytical technique and 0.10 M perchloric acid as supporting electrolyte. It has been shown that the presence of PoAP film increases considerably the efficiency of deposited Pt microparticles toward the oxidation of methanol. The catalytic activity of Pt particles is further enhanced when Ru or specially Sn is co‐deposited in the polymer film. The effects of various parameters such as the thickness of polymer film, concentration of methanol, medium temperature as well as the long term stability of modified electrodes have also been investigated.     

Transformation of Cell‐Derived Microparticles into Quantum‐Dot‐Labeled Nanovectors for Antitumor siRNA Delivery

Cell‐derived microparticles (MPs) have been recently recognized as critical intercellular information conveyors. However, further understanding of their biological behavior and potential application has been hampered by the limitations of current labeling techniques. Herein, a universal donor‐cell‐assisted membrane biotinylation strategy was proposed for labeling MPs by skillfully utilizing the natural membrane phospholipid exchange of their donor cells. This innovative strategy conveniently led to specific, efficient, reproducible, and biocompatible quantum dot (QD) labeling of MPs, thereby reliably conferring valuable traceability on MPs. By further loading with small interference RNA, QD‐labeled MPs that had inherent cell‐targeting and biomolecule‐conveying ability were successfully employed for combined bioimaging and tumor‐targeted therapy. This study provides the first reliable and biofriendly strategy for transforming biogenic MPs into functionalized nanovectors.     

Highly porous poly(lactide‐co‐glycolide) microparticles for sustained tiotropium release

In this study, porous poly(lactide‐co‐glycolide) (PLGA) microparticles with low mass density and large particle size were developed for chronic obstructive pulmonary disease treatment using anticholinergic drug (tiotropium). The porous PLGA microparticles were prepared by the water‐in‐oil‐in‐water (W₁/O/W₂) multi‐emulsion method using PLGA polymer and ammonium bicarbonate (as a porogen). Herein, soluble starch was incorporated in porous PLGA microparticles for long‐term tiotropium release. In vitro drug release studies determined that the rapid release of tiotropium from porous PLGA microparticles was reduced because of the high viscosity of the incorporated starch. Tiotropium release from porous PLGA microparticles continued up to 3 days. Furthermore, the inhaled microparticles showed longer drug residence in in vivo lung epithelium. Copyright © 2013 John Wiley & Sons, Ltd.     

Compartmentalized Photoreactions within Compositionally Anisotropic Janus Microstructures

We demonstrate spatially controlled photoreactions within bicompartmental microparticles and microfibers. Selective photoreactions are achieved by anisotropic incorporation of photocrosslinkable poly(vinyl cinnamate) in one compartment of either colloids or microfibers. Prior to photoreaction, bicompartmental particles, and fibers were prepared by EHD co‐jetting of two compositionally distinct polymer solutions. Physical and chemical anisotropy was confirmed by confocal laser scanning microscopy, Fourier‐transformed infrared spectroscopy, and scanning electron microscopy. The data indicate adjustment of polymer concentrations of the jetting solutions to be the determining factors for particle and fiber structures. Subsequent exposure of poly(vinyl cinnamate)‐based particles and fibers to UV light at 254 nm resulted in spatially controlled crosslinking. Treatment of the crosslinked bicompartmental colloids with chloroform produced half‐moon shaped objects. These hemishells exhibited a distinct porous morphology with pore sizes in the range of 70 nm. Based on this novel synthetic approach, Janus‐type particles and fibers can be prepared by EHD co‐jetting and can be selectively photocrosslinked without the need for masks or selective laser writing.

     

Reduction in microparticle adsorption using a lateral interconnection method in a PDMS‐based microfluidic device

Microparticle adsorption on microchannel walls occurs frequently due to nonspecific interactions, decreasing operational performance in pressure‐driven microfluidic systems. However, it is essential for delicate manipulation of microparticles or cells to maintain smooth fluid traffic. Here, we report a novel microparticle injection technique, which prevents particle loss, assisted by sample injection along the direction of fluid flow. Sample fluids, including microparticles, mammalian (U937), and green algae (Chlorella vulgaris) cells, were injected directly via a through hole drilled in the lateral direction, resulting in a significant reduction in microparticle attachment. For digital microfluidic application, the proposed regime achieved a twofold enhancement of single‐cell encapsulation compared to the conventional encapsulation rate, based on a Poisson distribution, by reducing the number of empty droplets. This novel interconnection method can be straightforwardly integrated as a microparticle or cell injection component in integrated microfluidic systems.     

Sol Processing of Conjugated Carbon Nitride Powders for Thin‐Film Fabrication

The chemical protonation of graphitic carbon nitride (CN) solids with strong oxidizing acids, for example HNO₃, is demonstrated as an efficient pathway for the sol processing of a stable CN colloidal suspension, which can be translated into thin films by dip/disperse‐coating techniques. The unique features of CN colloids, such as the polymeric matrix and the reversible hydrogen bonding, result in the thin‐film electrodes derived from the sol solution exhibiting a high mechanical stability with improved conductivity for charge transport, and thus show a remarkably enhanced photo‐electrochemical performance. The polymer system can in principle be broadly tuned by hybridization with desired functionalities, thus paving the way for the application of CN for specific tasks, as exemplified here by coupling with carbon nanotubes.     

Direct Surface‐Enhanced Raman Scattering Analysis of DNA Duplexes

The exploration of the genetic information carried by DNA has become a major scientific challenge. Routine DNA analysis, such as PCR, still suffers from important intrinsic limitations. Surface‐enhanced Raman spectroscopy (SERS) has emerged as an outstanding opportunity for the development of DNA analysis, but its application to duplexes (dsDNA) has been largely hampered by reproducibility and/or sensitivity issues. A simple strategy is presented to perform ultrasensitive direct label‐free analysis of unmodified dsDNA with the means of SERS by using positively charged silver colloids. Electrostatic adhesion of DNA promotes nanoparticle aggregation into stable clusters yielding intense and reproducible SERS spectra at nanogram level. As potential applications, we report the quantitative recognition of hybridization events as well as the first examples of SERS recognition of single base mismatches and base methylations (5‐methylated cytosine and N6‐methylated Adenine) in duplexes.     

Composite Hydrogels for Sustained Release of Therapeutic Agents

The goal of this research is to develop a composite hydrogel system for sustained release of therapeutic agents. The hydrogel composites were prepared by embedding drug‐loaded, biodegradable poly (DL‐lactide‐co‐glycolide) (PLGA) microparticles in semicrystalline hydrogels of polyvinyl alcohol (PVA). The gels were physically cross‐linked by the formation of the crystallites. The presence of the crystallites and the composite nature of the structure were confirmed by using differential scanning calorimetry and ATR‐FTIR spectroscopy. The distribution of microparticles in the hydrogel matrix was evaluated by using confocal laser scanning microscopy with coumarin‐6 as a fluorescence marker. The numbers of particles in the hydrogel matrix increased along the scanning depth, indicating uneven distribution. The release behavior of a model therapeutic agent, hydrocortisone, was evaluated, and the hydrogel composite system provided for better control of release than the microparticles and hydrogels alone. The addition of outer layers of PVA to the original single‐layer composite further reduced the initial burst effect from the microparticles and allowed for a linear release profile for greater than 1 month.     

Hollow Cu‐NP Spheres Made from Electroless Cu Deposition with Colloidal Particles as Templates

Summary: Hollow copper nanoparticle (Cu‐NP) spheres with different wall thicknesses were prepared with organic and inorganic colloids as core combining self‐assembly and electroless Cu deposition techniques. The hollow Cu‐NP spheres made from polymeric colloids possessed a thicker wall but were easy to break in the removal of the core by THF, while those from SiO₂ colloids had thinner wall and remained intact in the removal of the core by HF.

TEM image of the hollow Cu‐NP spheres.     

Particle‐in‐a‐Frame Nanostructures with Interior Nanogaps

Designing plasmonic hollow colloids with small interior nanogaps would allow structural properties to be exploited that are normally linked to an ensemble of particles but within a single nanoparticle. Now, a synthetic approach for constructing a new class of frame nanostructures is presented. Fine control over the galvanic replacement reaction of Ag nanoprisms with Au precursors gave unprecedented Au particle‐in‐a‐frame nanostructures with well‐defined sub‐2 nm interior nanogaps. The prepared nanostructures exhibited superior performance in applications, such as plasmonic sensing and surface‐enhanced Raman scattering, over their solid nanostructure and nanoframe counterparts. This highlights the benefit of their interior hot spots, which can highly promote and maximize the electric field confinement within a single nanostructure.     

Particle‐Stabilized Water Droplets that Sprout Millimeter‐Scale Tubes

Millimeter‐scale tubes are observed to sprout from water droplets injected into a bath of toluene containing ethanol and silica colloids. This phenomenon requires that first a membrane is formed by the colloids which self‐assemble at the droplet interface, and second, that the ethanol preferentially partitions into the aqueous phase leading to an internal over‐pressure. Tube growth, eruption, and shuffling droplets are subsequently observed, depending on the concentration of ethanol and colloids selected. This work opens many possibilities in the field of biomimetic droplets for fundamental studies of artificial growth at the microscale and for emulsion‐related applications.     

Ultra‐Stable Plasmonic Colloidal Aggregates for Accurate and Reproducible Quantitative SE(R)RS in Protein‐Rich Biomedia

Au/Ag colloids aggregated with simple salts are amongst the most commonly used substrates in surface‐enhanced (resonance) Raman spectroscopy (SE(R)RS). However, salt‐induced aggregation is a dynamic process, which means that SE(R)RS enhancements vary with time and that measurements therefore need to be taken at a fixed time point, normally within a short time‐window of a few minutes. Here, we present an emulsion templated method which allows formation of densely‐packed quasi‐spherical Au/Ag colloidal aggregates. Since the particles in the product aggregates retain their weakly adsorbed charged ligands and the ionic strength remains low these charged aggregates resist further aggregation while still providing intense SE(R)RS enhancement which remains stable for days. This eliminates a major source of irreproducibility in conventional colloidal SE(R)RS measurements and paves the way for SE(R)RS analysis in complex systems, such as protein‐rich bio‐solutions where conventional aggregated colloids fail.