On-chip fabrication and in-flow 3D-printing of microgel constructs: from chip to scaffold materials in one integral process.

Bioprinting has evolved into a thriving technology for the fabrication of cell-laden scaffolds. Bioinks are the most critical component for bioprinting. Recently, microgels have been introduced as a very promising bioink, enabling cell protection and the control of the cellular microenvironment. However, the fabrication of the bioinks involves the microfluidic production of the microgels, with a subsequent multistep process to obtain the bioink, which so far has limited its application potential. Here we introduce a direct coupling of microfluidics and 3D-printing for the continuous microfluidic production of microgels with direct in-flow printing into stable scaffolds. The 3D-channel design of the microfluidic chip provides access to different hydrodynamic microdroplet formation regimes to cover a broad range of droplet and microgel diameters. After exiting a microtubing the produced microgels are hydrodynamically jammed into thin microgel filaments for direct 3D-printing into two- and three-dimensional scaffolds. The methodology enables the continuous on-chip encapsulation of cells into monodisperse microdroplets with subsequent in-flow cross-linking to produce cell-laden microgels. The method is demonstrated for different cross-linking methods and cell lines. This advancement will enable a direct coupling of microfluidics and 3D-bioprinting for scaffold fabrication.

Bistability, Remanence, Read/Write-Memory, and Logic Gate Function via a Stimuli-Responsive Polymer.

Stimuli-responsive materials change their state in response to external triggers. Switching between different states enables information to be written, stored, and read, if the transition between the states exhibits hysteresis. Thermally responsive polymers exhibit an intrinsic hysteresis for the volume phase transition between the swollen and de-swollen solution state. Here, it is shown that this hysteresis can be used to realize bistability, remanence, and reversible write/read information storage. This is demonstrated for the simplest and most widely used thermoresponsive polymer, poly(N-isopropylacrylamide) (PNIPAM), as well as for PNIPAM block copolymers, which widens the hysteresis window. The hysteresis is shown to be related to cluster domain assembly/disassembly during the phase transition. Information can be written thermally using a laser, or using heated or cooled pen tips on a thin-film backscattering display. The bistable state can additionally be switched by pH, enabling an AND logic gate function. Furthermore, an unusual memory state is discovered, where information is visible in the hysteresis window and invisible at higher temperatures, allowing encoded information to be hidden. Since hysteresis is a very common intrinsic phenomenon for responsive materials, this principle to encode and store information is potentially applicable to a broad range of responsive materials.

Simultaneous SAXS/WAXS/UV-Vis Study of the Nucleation and Growth of Nanoparticles: A Test of Classical Nucleation Theory.

Despite the increasing interest in the applications of functional nanoparticles, a comprehensive understanding of the formation mechanism starting from the precursor reaction with subsequent nucleation and growth is still a challenge. We for the first time investigated the kinetics of gold nanoparticle formation systematically by means of a lab-based in situ small-angle X-ray scattering (SAXS)/wide-angle X-ray scattering (WAXS)/UV-vis absorption spectroscopy experiment using a stopped-flow apparatus. We thus could systematically investigate the influence of all major factors such as precursor concentration, temperature, the presence of stabilizing ligands and cosolvents on the temporal evolution of particle size, size distribution, and optical properties from the early prenucleation state to the late growth phase. We for first time formulated and numerically solved a closed nucleation and growth model including the precursor reaction. We observe that the results can be well described within the framework of classical nucleation and growth theory, including also results of previous studies by other research groups. From the analysis, we can quantitatively derive values for the rate constants of precursor reaction and growth together with their activation free enthalpies. We find the growth process to be surface-reaction limited with negligible influence of Ostwald ripening yielding narrow disperse gold nanoparticles.

Thermal ink-jet spray freeze-drying for preparation of excipient-free salbutamol sulphate for inhalation.

The use of thermal ink-jet spray freeze-drying (TIJ-SFD) to engineer inhalable, excipient-free salbutamol sulphate (SS) particles was assessed. A modified Hewlett-Packard printer was used to atomise aqueous SS solutions into liquid nitrogen. The frozen droplets were freeze-dried. It was found that TIJ-SFD could process SS solutions up to 15%w/v; the porous particles produced had a physical diameter of ca. 35 µm. Next generation impactor (NGI) analysis indicated that the particles had a smaller aerodynamic size (MMAD ranging from 6 to 8.7 µm). Particles prepared from the lowest concentration SS solution were too fragile to withstand aerosolisation, but the 5%w/v solution yielded particles having the best combination of strength and aerodynamic properties. Comparison with a commercial SS formulation (Cyclocap®) showed that the SFD preparation had an almost equivalent FPF (6.4 µm) when analysed with a twin-stage impinger (TSI; 24.0 ± 1.2% and 26.4 ± 2.2%, respectively) and good performance when analysed with NGI (FPF (4.46 µm):16.5 ± 2.0 and 27.7 ± 1.7, respectively). TIJ-SFD appears to be an excellent method to prepare inhalable particles. It is scalable yet allows assessment of the viability of the pulmonary route early in the development since it can be used with very small volumes (<0.5 mL) of solution.

Preparation of monodisperse block copolymer vesicles via a thermotropic cylinder-vesicle transition.

In aqueous solution, poly(2-vinylpyridine-b-ethylene oxide) spontaneously forms bilayer vesicles, the size of which can be tailored by extrusion through polycarbonate membranes. However, their size can be even more precisely influenced by subjecting them to a specific cooling/warming process proceeding through a cylinder-vesicle shape transition. The thermotropic alterations of the polymer aggregates and the topological pathways of the cylinder-vesicle transition were followed by dynamic light scattering (DLS) and cryo-electron microscopy (cryo-TEM). Upon cooling the vesicles to 4degreesC, there is a transition of the vesicles to basketlike aggregates and their further disintegration to wormlike micelles. Rewarming of the dispersion results in the reformation of vesicles via intermediate discoid and octopus-like structures. The variation of incubation times at 4 and 25degreesC, heating rate, polymer concentration, and ionic strength allows tailored preparation of unilamellar and almost monodisperse vesicles with diameters between 60 and 500 nm. Furthermore, fluorescently labeled dextrans, which were used as model drugs of differing molar mass, could be easily and stably encapsulated during the thermotropic formation of vesicles from wormlike micelles.

Characteristics of picoliter droplet dried residues as standards for direct analysis techniques.

The characteristics of dried residues of picodroplets of single-, two-, and three-element aqueous solutions, which qualify these as reference materials in the direct analysis of single particles, single cells, and other microscopic objects using, e.g., laser ablation inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-TOF-MS) and micro-X-ray fluorescence (MXRF), were evaluated. Different single-, two-, and three-element solutions (0.01-1 g/L) were prepared in picoliter volume (around 130 pL) with a thermal inkjet printing technique. An achievable dosing precision of 4-15% was calculated by total reflection X-ray fluorescence (TXRF) determination of the transferred elemental mass of an array of 100 droplets. The size of the dried residues was determined by optical microscopy to be 5-20 microm in diameter depending on the concentration and the surface material. The elemental distribution of the dried residues was determined with synchrotron micro-X-ray fluorescence (SR-MXRF) analyses. The MXRF results show high uniformity for element deposition of every single droplet with an RSTD of 4-6% depending on the concentration of spotted solution. The shape and height profile of dried residues from picoliter droplets were studied using atomic force microscopy (AFM). It was found that these dry to give symmetrical spherical segments with maximum heights of 1.7 microm. The potential of this technique for direct LA-ICP-TOF-MS analysis is shown.

Molecular exchange through membranes of poly(2-vinylpyridine-block-ethylene oxide) vesicles.

The molecular exchange of tracer molecules through the membranes of dispersed vesicles of the block copolymer poly(2-vinylpyridine-block-ethylene oxide) was studied by using NMR spectroscopy combined with pulsed field gradients. The hydrodynamic radius of the tracer molecules was varied systematically to obtain a permeability profile of the vesicle membrane. In addition, the effect of system parameters, such as temperature, pH value, vesicle size, and thickness of the vesicle membrane, was studied. In the case of rapid exchange with average residence times significantly smaller than 10 s, the permeation is observed under equilibrium conditions and the data are analyzed by using a simple analytical approach. For slow exchange processes with average residence times above 10 s, the permeation is monitored in a time-resolved measurement under nonequilibrium conditions. Generally, the transmembrane exchange rate of the tracer clearly depends on its hydrodynamic radius. The characteristics of this dependence indicate the presence of two different mechanisms of membrane penetration, one dominating for smaller and one for larger tracer molecules, respectively. The exchange rate also shows a significant dependence on the bilayer thickness and on the vesicle diameter. By contrast, no variation of the membrane permeability with the temperature or the pH value could be detected as long as the vesicles remain stable.