Strategies for the Biofunctionalization of Straining Flow Spinning Regenerated Bombyx mori Fibers.

High-performance regenerated silkworm (Bombyx mori) silk fibers can be produced efficiently through the straining flow spinning (SFS) technique. In addition to an enhanced biocompatibility that results from the removal of contaminants during the processing of the material, regenerated silk fibers may be functionalized conveniently by using a range of different strategies. In this work, the possibility of implementing various functionalization techniques is explored, including the production of fluorescent fibers that may be tracked when implanted, the combination of the fibers with enzymes to yield fibers with catalytic properties, and the functionalization of the fibers with cell-adhesion motifs to modulate the adherence of different cell lineages to the material. When considered globally, all these techniques are a strong indication not only of the high versatility offered by the functionalization of regenerated fibers in terms of the different chemistries that can be employed, but also on the wide range of applications that can be covered with these functionalized fibers.

Scaling Laws of an Exploding Liquid Column under an Intense Ultrashort X-Ray Pulse.

A general formulation of the partial destruction of a liquid object in vacuum after the sudden deposition of a very large amount of energy is proposed. That energy instantaneously raises the pressure of a portion of the liquid to extreme values and changes its state, which causes its explosive expansion into vacuum and against the rest of the liquid object. When the deformable object is a liquid capillary column, the model reduces to a universal equation for the evolution of the expanding gap between the two sides of the exploding liquid column. The theoretical analysis contemplates two asymptotic stages for small and large times from the initiation of the blast, whose asymptotic solutions are fitted to available experimental data. A universal approximate analytical solution is obtained. A complete dimensional analysis of the problem and an optimal collapse of experimental data reveal that the proposed solution is in remarkable agreement with experiments of a jet exploding after being irradiated by an ultrashort and intense x-ray pulse from an x-ray free electron laser.

Controlled cavity collapse: scaling laws of drop formation.

The formation of transient cavities at liquid interfaces occurs in an immense variety of natural processes, among which the bursting of surface bubbles and the impact of a drop on a liquid pool are salient. The collapse of a surface liquid cavity is a well documented natural process that leads to the ejection of a thin and fast jet. Droplets generated through this process can be one order of magnitude smaller than the cavity's aperture, and they are consequently of interest in drop on demand inkjet applications. In this work, the controlled formation and collapse of a liquid cavity is analyzed, and the conditions for minimizing the resulting size and number of ejected drops are determined. The experimental and numerical models are simple and consist of a liquid reservoir, a nozzle plate with the discharge orifice, and a moving piston actuated by single half-sine-shaped pull-mode pulses. The size of the jetted droplet is described by a physical model resulting in a scaling law that is numerically and experimentally validated.

Revision of Bubble Bursting: Universal Scaling Laws of Top Jet Drop Size and Speed.

The collapse of a bubble of radius R_{o} at the surface of a liquid generating a liquid jet and a subsequent first drop of radius R is universally scaled using the Ohnesorge number Oh=µ/(ρσR_{o})^{1/2} and a critical value Oh^{*} below which no droplet is ejected; ρ, σ, and µ are the liquid density, surface tension, and viscosity, respectively. First, a flow field analysis at ejection yields the scaling of R with the jet velocity V as R/l_{µ}∼(V/V_{µ})^{-5/3}, where l_{µ}=µ^{2}/(ρσ) and V_{µ}=σ/µ. This resolves the scaling problem of curvature reversal, a prelude to jet formation. In addition, the energy necessary for the ejection of a jet with a volume and averaged velocity proportional to R_{o}R^{2} and V, respectively, comes from the energy excess from the total available surface energy, proportional to σR_{o}^{2}, minus the one dissipated by viscosity, proportional to µ(σR_{o}^{3}/ρ)^{1/2}. Using the scaling variable φ=(Oh^{*}-Oh)Oh^{-2}, it yields V/V_{µ}=k_{v}φ^{-3/4} and R/l_{µ}=k_{d}φ^{5/4}, which collapse published data since 1954 and resolve the scaling of R and V with k_{v}=16, k_{d}=0.6, and Oh^{*}=0.043 when gravity effects are negligible.

Production of High Performance Bioinspired Silk Fibers by Straining Flow Spinning.

In the last years, there has been an increasing interest in bioinspired approaches for different applications, including the spinning of high performance silk fibers. Bioinspired spinning is based on the natural spinning system of spiders and worms and requires combining changes in the chemical environment of the proteins with the application of mechanical stresses. Here we present the novel straining flow spinning (SFS) process and prove its ability to produce high performance fibers under mild, environmentally friendly conditions, from aqueous protein dopes. SFS is shown to be an extremely versatile technique which allows controlling a large number of processing parameters. This ample set of parameters allows fine-tuning the microstructure and mechanical behavior of the fibers, which opens the possibility of adapting the fibers to their intended uses.

Effect of a Surrounding Liquid Environment on the Electrical Disruption of Pendant Droplets.

The effect of a surrounding, dielectric, liquid environment on the dynamics of a suddenly electrified liquid drop is investigated both numerically and experimentally. The onset of stability of the droplet is naturally dictated by a threshold value of the applied electric field. While below that threshold the droplet retains its integrity, reaching to a new equilibrium state through damped oscillations (subcritical regime), above it electrical disruption takes place (supercritical regime). In contrast to the oscillation regime, the dynamics of the electric droplet disruption in the supercritical regime reveals a variety of modes. Depending on the operating parameters and fluid properties, a drop in the supercritical regime may result in the well-known tip streaming mode (with and without whipping instability), in droplet splitting (splitting mode), or in the development of a steep shoulder at the elongating front of the droplet that expands radially in a sort of "splashing" (splashing mode). In both splitting and splashing modes, the sizes of the progeny droplets, generated after the breakup of the mother droplet, are comparable to that of the mother droplet. Furthermore, the development in the emission process of the shoulder leading to the splashing mode is described as a parametrical bifurcation, and the parameter governing that bifurcation has been identified. Physical analysis confirms the unexpected experimental finding that the viscosity of the dynamically active environment is absent in the governing parameter. However, the appearance of the splitting mode is determined by the viscosity of the outer environment, when that viscosity overcomes a certain large value. These facts point to the highly nonlinear character of the drop fission process as a function of the droplet volume, inner and outer liquid viscosities, and applied electric field. These observations may have direct implications in systems where precise control of the droplet size is critical, such as in analytical chemistry and "drop-on-demand" processes driven by electric fields.

The high-throughput atomization of polymer solutions for fiber synthesis in a single step aided with corona ionizers.

Polymer microfibers are ubiquitous structures across virtually all technological fields. Their applications include, for instance, filter media, tissue regeneration, wound healing and dressing, and reinforcement materials. The most effective methods for fabrication of fibrous micro and nanomaterials rely on electric fields to spin a liquid jet into an ultrafine thread that rapidly dries up forming a fiber. Continuous spinning and collection leads to formation of fiber mats. Here we report a robust yet simple approach for the massive production of liquid threads, which upon acquiring electrical charges in-flight are collected downstream in the form of fibers. The entire process takes place on-line in a single step. The liquid threads are produced through the fragmentation of a polymer solution bulk due to a turbulent interaction of a gas-liquid interface in the interior of an engineered device, a so-called Flow Blurring atomizer. The particularity of this approach consists precisely in such vigorous interaction, at the micrometer scale, which triggers a bubbly motion in the interior of the device, that is a "micro-mixing". Subsequently, the threads are passed through ionized air currents, at ambient conditions, and then stretched to sub-micrometer dimensions by electric fields. Polyvinylpyrrolidone (PVP) as well as carbon nanotubes (CNTs) or graphene oxide sheets (GOSs)-containing PVP fibers, with diameters in the range 100-900 nm, were synthesized via this approach. In the cases studied herein the method was operated at liquid flow rates (i.e. production rates) of 0.2 mL/min but it could be readily increased up to a few tens of mL/min. The method requires further improvement and optimization, nevertheless it is a promising alternative for mass production of polymer fibers.

Integrating a micro-mixing mechanism and on-line thermal processing for the large-scale ejection of polymeric liquid threads for producing ultrafine fibers.

Micro/nanofibers are structures that nowadays have a wide range of cutting-edge applications including energy generation and storage devices, smart textiles, cell growth, and tissue engineering. These fibrous materials are mostly produced from polymer solutions spun, under laminar flow conditions, into nanofibers by external forces. However, the turbulent interaction of gas-liquid interfaces offers an innovative approach for the high-throughput production of nanofibers. Here, we present Flow Blurring (FB), a solely pneumatic approach for the massive production of liquid threads of polymer solutions, which relies on a micro-mixing mechanism that triggers a turbulent motion capable of fragmenting a viscous flow. The as-ejected threads are subsequently processed thermally, on-line in a single-step, thus producing micro/nanofibers that form mats. The method operates with relatively large liquid flow rates, equivalent of a high production rate, and is thus suitable for industrial production of engineered nanomaterials. In this work, we used solutions of poly(vinyl alcohol) (PVA) to study its ejection and fragmentation dynamics through computational fluid dynamics (CFD) simulations. In addition, the physics underlying the regulation of the liquid flow rate in FB atomizers are proposed. Fibers with typical diameters in the range 400-800 nm were produced by online heating of the liquid threads. Liquid ejection experiments were performed under different operating conditions thus verifying the capability of the method for synthesizing submicrometer-sized fibers with high uniformity and production rates suitable for scaling up.

An experimental study of liquid micro-jets produced with a gas dynamic virtual nozzle under the influence of an electric field.

The results of an experimental study of micro-jets produced with a gas dynamic virtual nozzle (GDVN) under the influence of an electric field are provided and discussed for the first time. The experimental study is performed with a 50% volume mixture of water and ethanol, and nitrogen focusing gas. The liquid sample and gas Reynolds numbers range from 0.09-5.4 and 0-190, respectively. The external electrode was positioned 400-500 µm downstream of the nozzle tip and an effect of electric potential between the electrode and the sample liquid from 0-7 kV was investigated. The jetting parametric space is examined as a function of operating gas and liquid flow rates, outlet chamber pressure, and an external electric field. The experimentally observed jet diameter, length and velocity ranged from 1-25 µm, 50-500 µm and 0.5-10 m/s, respectively. The jetting shape snapshots were processed automatically using purposely developed computer vision software. The velocity of the jet was calculated from the measured jet diameter and the sample flow rate. It is found that micro-jets accelerate in the direction of the applied electric field in the downstream direction at a constant acceleration as opposed to the standard GDVNs. New jetting modes were observed, where either the focusing gas or the electric forces dominate, encouraging further theoretical and numerical studies towards optimized system design. The study shows the potential to unlock a new generation of low background sample delivery for serial diffraction measurements of weakly scattering objects.

Massive production of fibroin nano-fibrous biomaterial by turbulent co-flow.

Among the different polymers (proteins, polysaccharides, etc.) that make up natural fibers, fibroin is a protein produced by silk spinning animals, which have developed an optimized system for the conversion of a highly concentrated solution of this protein into high-performance solid fibers. This protein undergoes a self-assembly process in the silk glands that result from chemical gradients and by the application of mechanical stresses during the last step of the process. In the quest for a process that could mimic natural spinning at massive scales, we have discovered that turbulence offers a novel and promising solution: a turbulent liquid jet can be formed by a chemically green and simple coagulating liquid (a diluted solution of acetic acid in etanol) co-flowing with a concentrated solution of fibroin in water by the use of a Flow Blurring nebulizer. In this system, (a) the co-flowing coagulant liquid extracts water from the original protein solution and, simultaneously, (b) the self-assembled proteins are subjected to mechanical actions, including splitting and stretching. Given the non-negligible produced content with the size and appearance of natural silk, the stochastic distribution of those effects in our process should contain the range of natural ones found in animals. The resulting easily functionalizable and tunable one-step material is 100% biocompatible, and our method a perfect candidate to large-scale, low-cost, green and sustainable processing of fibroin for fibres and textiles.

Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers.

The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX.

On the Ejection of Filaments of Polymer Solutions Triggered by a Micrometer-Scale Mixing Mechanism.

Polymer filaments constitute precursor materials of so-called fiber mats, ubiquitous structures across cutting-edge technological fields. Thus, approaches that contribute to large-scale production of fibers are desired from an industrial perspective. Here, we use a robust liquid atomization device operated at relatively high flow rates, ~20 mL/min, as facilitating technology for production of multiple polymer filaments. The method relies on a turbulent, energetically efficient micro-mixing mechanism taking place in the interior of the device. The micro-mixing is triggered by radial implosion of a gas current into a liquid feeding tube, thus resulting in breakup of the liquid surface. We used poly(ethylene oxide) solutions of varying concentrations as test liquids to study their fragmentation and ejection dynamics employing ultra-high speed imaging equipment. Taking an energy cascade approach, a scaling law for filament diameter was proposed based on gas pressure, liquid flow rate and viscosity. We find that a filament dimensionless diameter, Df*, scales as a non-dimensional liquid flow rate Q* to the 1/5. The study aims to elucidate the underlying physics of liquid ejection for further applications in material production.

Regenerated Silk Fibers Obtained by Straining Flow Spinning for Guiding Axonal Elongation in Primary Cortical Neurons.

The recovery of injured nervous tissue, one of the main goals for regenerative therapeutic approaches, is often hindered by the limited axonal regeneration ability of the central nervous system (CNS). In this regard, the identification of scaffolds that support the reconstruction of functional neuronal tissues and guide the alignment of regenerating neurons is a major challenge in tissue engineering. Ideally, the usage of such scaffolds would promote and guide the axonal growth, a crucial phase for the restoration of neuronal connections and, consequently, the nerve function. Among the materials proposed as scaffolds for CNS regeneration, silk has been used to exploit its outstanding features as a biomaterial to promote axonal regeneration. In this study, we explore, for the first time, the possibility of using high-performance regenerated silk fibers obtained by straining flow spinning (SFS) to serve as scaffolds for inducing and guiding the axonal growth. It is shown that SFS fibers promote the spontaneous organization of dissociated cortical primary cells into highly interconnected cellular spheroid-like tissue formations. Neuronal projections (i.e., axons) from these cellular spheroids span hundreds of microns along the SFS fibers that act as guides and allow the connection of distant spheroids. In addition, it is also shown that SFS fibers serve as scaffolds for neuronal migration covering short and long distances. As a consequence, the usage of high-performance SFS fibers appears as a promising basis for the development of novel therapies, leading to directed axonal regeneration.

Revision of capillary cone-jet physics: electrospray and flow focusing.

Capillary cone jets are natural microfluidic structures arising in steady capillary tip streaming, whose paradigms are electrospray and flow focusing phenomena. In this work, we make a profound revision of the basic underlying physics of generic cone jets from thousands of experimental measurements, most of them reported in the literature. First, the boundaries of the stability region of steady jetting are calculated. We describe these limitations by instability mechanisms associated with the local flow structure in the tip and the issuing jet and with the global behavior of the meniscus. Second, to undertake a general physical treatment of cone jets in steady regime, we analyze the energy balance taking place in the tips of both flow focusing and electrospray. This analysis yields a fundamental result: if the electrospray data are expressed in terms of an effective pressure drop, both phenomena satisfy the same scaling law for the droplet size, which exhibits nearly complete similarity in the parameter window where quasimonodisperse sprays are produced. That effective pressure drop is a function of the liquid properties exclusively, i.e., it does not depend on the operational parameters (flow rate and applied voltage). Moreover, the stability limits of the operational regimes are analyzed in detail, finding fundamental coincidences between flow focusing and electrospray as well. These results provide most useful general description and predictive scaling laws for nearly monodisperse microspraying or nanospraying based on steady cone jets, of immediate applicability in analytical chemistry, chemical engineering, biochemistry, pharmaceutical and food technologies, painting, and many other technological fields.

Flow Blurring-Enabled Production of Polymer Filaments from Poly(ethylene oxide) Solutions.

Flow blurring (FB) atomizers are relatively simple yet robust devices used for the generation of sprays from solutions of a wide range of viscosities. In this work, we have demonstrated that FB devices may also be applied for massive production of liquid filaments from polymeric solutions. They can later be transformed into solid filaments and fibers, leading to the production of so-called fiber mats. The liquid precursors consisted of poly(ethylene oxide) (PEO) solutions of varying molecular weights (105 [100k] to 4 × 106 g/mol [4M]) and concentrations. The FB device was operated in the gas pressure range of 3-6 bar. Except for solutions of PEO 100k, all solutions exhibited a shear thinning behavior. For massive filament production, a threshold polymer concentration (c t) was identified for each molecular weight. Below such concentration, the atomization resulted in droplets (the classical FB functioning mode). Such a threshold value decreased as the PEO molecular weight increased, and it coincides with the polymer coil overlap concentration, c*. The viscoelastic nature of the solutions was also observed to increase with the molecular weight. A 3.2 dependency of the zero-shear rate viscosity on a so-called Bueche parameter was found for filament production, whereas a nearly linear dependency was found for droplet production. In general, the mean diameter of the filaments decreased as they traveled downstream from the atomization point. Furthermore, at a given distance from the atomizer outlet and gas pressure, the mean filament diameter slightly shifted toward larger sizes with increasing PEO molecular weight. The tendency agrees well with the calculated filaments' Deborah number, which increases with PEO molecular weight. The approach presented herein describes a high-throughput and efficient method for the massive production of viscous filaments. These may be transformed into fibers by an on-line drying step.

Emergence of supercontraction in regenerated silkworm (Bombyx mori) silk fibers.

The conditions required for the emergence of supercontraction in regenerated silkworm (Bombyx mori) silk fibers are assessed through an experimental approach that combines the spinning of regenerated fibers with controlled properties and their characterization by 13C solid-state nuclear magnetic resonance (NMR). Both supercontracting and non-supercontracting regenerated fibers are produced using the straining flow spinning (SFS) technique from 13C labeled cocoons. The short-range microstructure of the fibers is assessed through 13C CP/MAS in air and 13C DD/MAS in water, and the main microstructural features are identified and quantified. The mechanical properties of the regenerated fibers and their microstructures are compared with those of natural silkworm silk. The combined analysis highlights two possible key elements as responsible for the emergence of supercontraction: (1) the existence of an upper and a lower limit of the amorphous phase compatible with supercontraction, and (2) the existence of two ordered phases, ß-sheet A and B, which correspond to different packing arrangements of the protein chains.

Evaluation of serial crystallographic structure determination within megahertz pulse trains.

The new European X-ray Free-Electron Laser (European XFEL) is the first X-ray free-electron laser capable of delivering intense X-ray pulses with a megahertz interpulse spacing in a wavelength range suitable for atomic resolution structure determination. An outstanding but crucial question is whether the use of a pulse repetition rate nearly four orders of magnitude higher than previously possible results in unwanted structural changes due to either radiation damage or systematic effects on data quality. Here, separate structures from the first and subsequent pulses in the European XFEL pulse train were determined, showing that there is essentially no difference between structures determined from different pulses under currently available operating conditions at the European XFEL.

Unconditional jetting.

Capillary jetting of a fluid dispersed into another immiscible phase is usually limited by a critical capillary number, a function of the Reynolds number and the fluid property ratios. Critical conditions are set when the minimum spreading velocity of small perturbations v_{-};{*} along the jet (marginal stability velocity) is zero. Here we identify and describe parametric regions of high technological relevance, where v_{-};{*}>0 and the jet flow is always supercritical independently of the dispersed liquid flow rate; within these relatively broad regions, the jet does not undergo the usual dripping-jetting transition, so that either the jet can be made arbitrarily thin (yielding droplets of any imaginably small size), or the issuing flow rate can be made arbitrarily small. In this work, we provide illustrative analytical studies of asymptotic cases for both negligible and dominant inertia forces. In this latter case, requiring a nonzero jet surface velocity, axisymmetric perturbation waves "surf" downstream for all given wave numbers, while the liquid bulk can remain static. In the former case (implying small Reynolds flow) we found that the jet profile small slope is limited by a critical value; different published experiments support our predictions.