Alignment-Rheology Relationship of Biosourced Rod-Like Colloids and Polymers under Flow.

Fluids composed of biosourced rod-like colloids (RC) and rod-like polymers (RP) have been extensively studied due to various promising applications relying on their flow-induced orientation (e.g., fiber spinning). However, the relationship between RC and RP alignment and the resulting rheological properties is unclear due to experimental challenges. We investigate the alignment-rheology relationship for a variety of biosourced RC and RP, including cellulose-based particles, filamentous viruses, and xanthan gum, by simultaneous measurements of the shear viscosity and fluid anisotropy under rheometric shear flows. For each system, the RC and RP contribution to the fluid viscosity, captured by the specific viscosity ηsp, follows a universal trend with the extent of the RC and RP alignment independent of concentration. We further exploit this unique rheological-structural link to retrieve a dimensionless parameter (ß) directly proportional to ηsp at zero shear rate (η0,sp), a parameter often difficult to access from experimental rheometry for RC and RP with relatively long contour lengths. Our results highlight the unique link between the flow-induced structural and rheological changes occurring in RC and RP fluids. We envision that our findings will be relevant in building and testing microstructural constitutive models to predict the flow-induced structural and rheological evolution of fluids containing RC and RP.

Microfluidic Assisted Nanoprecipitation of PLGA Nanoparticles for Curcumin Delivery to Leukemia Jurkat Cells.

The ability to control particle size and size distribution of nanoparticles for drug delivery is essential because it impacts on the biodistribution and cellular uptake of nanoparticles. We present a novel microfluidic assisted nanoprecipitation strategy that enables synthesis of surfactant-free curcumin encapsulated poly(lactide- co-glycolide) nanoparticles (Cur-PLGA NP) with adjustable particle diameters (30-70 nm) and narrow particle size distribution (polydispersity index less than 0.2). Our Cur-PLGA NP exhibit excellent colloidal stability and inhibit degradation of curcumin. We further demonstrate the potential of our Cur-PLGA NP as a nanotoxic delivery system for curcumin. Cellular viability assay validates a dose-dependent cytotoxicity of Cur-PLGA NP in leukemia Jurkat cells. In contrast, Cur-PLGA NP does not alter the viability of fibroblast NIH3T3 cells, which suggests that the cytotoxicity of Cur-PLGA NP is specific to cell types. Furthermore, there is no detectable effect by PLGA NP to both leukemia Jurkat cells and fibroblast NIH3T3 cells, highlighting the nontoxic nature of our delivery system. Confocal cell uptake studies indicate that PLGA NP do not alter the cell uptake of curcumin. Our microfluidic assisted approach offers a controlled and effective nanobiomaterials synthesis of drug delivery system for curcumin, which can be extended to different capsule materials for a variety of biomedical applications.

"From the Edge to the Center": Viscoelastic Migration of Particles and Cells in a Strongly Shear-Thinning Liquid Flowing in a Microchannel.

Controlling the fate of particles and cells in microfluidic devices is critical in many biomedical applications, such as particle and cell alignment and separation. Recently, viscoelastic polymer solutions have been successfully used to promote transversal migration of particles and cells toward fixed positions in straight microchannels. When inertia is negligible, numerical simulations have shown that strongly shear-thinning polymer solutions (fluids with a shear viscosity that decreases with increasing flow rates) promote transversal migration of particles and cells toward the corners or toward the centerline in a straight microchannel with a square cross section, as a function of particle size, cell deformability, and channel height. However, no experimental evidence of such shifting in the positions for particles or cells suspended in strongly shear-thinning liquids has been presented so far. In this work, we demonstrate that particle positions over the channel cross section can be shifted "from the edge to the center" in a strongly shear-thinning liquid. We investigate the viscoelasticity-induced migration of both rigid particles and living cells (Jurkat cells and NIH 3T3 fibroblasts) in an aqueous 0.8 wt % hyaluronic acid solution. The combined effect of fluid elasticity, shear-thinning, geometric confinement, and cell deformability on the distribution of the particle/cell positions over the channel cross section is presented and discussed. In the same shear-thinning liquid, separation of 10 and 20 µm particles is also achieved in a straight microchannel with an abrupt expansion. Our results envisage further applications in viscoelasticity-based microfluidics, such as deformability-based cell separation and viscoelastic spacing of particles/cells.

Flow-induced structured phase in nonionic micellar solutions.

In this work, we consider the flow of a nonionic micellar solution (precursor) through an array of microposts, with focus on its microstructural and rheological evolution. The precursor contains polyoxyethylene(20) sorbitan monooleate (Tween-80) and cosurfactant monolaurin (ML). An irreversible flow-induced structured phase (NI-FISP) emerges after the nonionic precursor flows through the hexagonal micropost arrays, when subjected to strain rates ~10(4) s(-1) and strain ~10(3). NI-FISP consists of close-looped micellar bundles and multiconnected micellar networks as evidenced by transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM). We also conduct small-angle neutron scattering (SANS) measurements in both precursor and NI-FISP to illustrate the structural transition. We propose a potential mechanism for the NI-FISP formation that relies on the micropost arrays and the flow kinematics in the microdevice to induce entropic fluctuations in the micellar solution. Finally, we show that the rheological variation from a viscous precursor solution to a viscoelastic micellar structured phase is associated with the structural evolution from the precursor to NI-FISP.

Microrheological Approach for Probing the Entanglement Properties of Polyelectrolyte Solutions.

The entanglement dynamics and viscoelasticity of polyelectrolyte solutions remain active research topics. Previous studies have reported conflicting experimental results when compared to Dobrynin's scaling predictions derived from the Doi-Edwards (DE) tube model for entangled polymers. Herein, by combining classical bulk shear rheometry with diffusing wave spectroscopy (DWS) microrheometry, we investigate how the key viscoelastic parameters (the specific viscosity ηsp, the plateau modulus Ge, and the ratio of the reptation time to the Rouse time of an entanglement strand τrep/τe) depend on the polymer concentration for semidilute entangled (SE) solutions containing poly(sodium styrenesulfonate) with high molecular weight. Our experimental measurements yield Ge ∝ c1.51±0.04, in good agreement with the scaling of Ge ∝ c1.5 predicted by Dobrynin's model for salt-free polyelectrolyte SE solutions, suggesting that the electrostatic interaction influences the viscoelastic properties of polyelectrolyte SE solutions. On the other hand, the deviation in the scaling exponent for ηsp ∝ c2.56±0.04 and τrep/τe ∝ c1.82±0.28 is observed between our DWS experiments and Dobrynin's model prediction (∝ c1.5), likely due to the fact that Dobrynin's scaling model does not account for mechanisms such as the contour length fluctuation, the constraint release, and the retardation of solvent dynamics, which are known to occur for SE solutions of neutral polymers. Our results demonstrate that DWS serves as a powerful rheological tool to study the entanglement dynamics of polyelectrolyte solutions. The scaling relationships obtained in this study provide new insights to the long-standing debate on the entanglement dynamics of polyelectrolyte solutions.

High-throughput fabrication of high aspect ratio Ag/Al nanopillars for optical detection of biomarkers.

Nanomaterial-based optical techniques for biomarker detection have garnered tremendous attention from the nanofabrication community due to their high precision and enhanced limit of detection (LoD) features. These nanomaterials are highly responsive to local refractive index (RI) fluctuations, and their RI unit sensitivity can be tuned by varying the chemical composition, geometry, and dimensions of the utilized nanostructures. To improve the sensitivity and LoD values of these nanomaterials, it is common to increase both dimensions and aspect ratios of the fabricated nanostructures. However, limited by the complexity, prolonged duration, and elevated costs of the available nanofabrication techniques, mass production of these nanostructures remains challenging. To address not only high accuracy, but also speed and production effectiveness in these nanostructures' fabrication, our work reports, for the first time, a fast, high-throughput, and cost-effective nanofabrication protocol for routine manufacturing of polymer-based nanostructures with high sensitivity and calculated LoD in the pM range by utilizing anodized aluminum oxide (AAO) membranes as templates. Specifically, our developed platform consists of arrays of nearly uniform polystyrene nanopillars with an average diameter of ∼185 nm and aspect ratio of ∼11. We demonstrate that these nanostructures can be produced at a high speed and a notably low price, and that they can be efficiently applied for biosensing purposes after being coated with aluminum-doped silver (Ag/Al) thin films. Our platform successfully detected very low concentrations of human C-reactive protein (hCRP) and SARS-CoV-2 spike protein biomarkers in human plasma samples with LoDs of 11 and 5 pM, respectively. These results open new opportunities for day-to-day fabrication of high aspect ratio arrays of nanopillars that can be used as a base for nanoplasmonic sensors with competitive LoD values. This, in turn, contributes to the development of point-of-care devices and further improvement of the existing nanofabrication techniques, thereby enriching the fields of pharmacology, clinical analysis, and diagnostics.

Structure-property relationship of a soft colloidal glass in simple and mixed flows.

HYPOTHESIS: Under specific conditions, rod-like cellulose nanocrystals (CNC) can assemble into structurally ordered soft glasses (SGs) with anisotropy that can be controlled by applying shear. However, to achieve full structural control of SGs in real industrial processes, their response to mixed shear and extensional kinematics needs to be determined. We hypothesise that by knowing the shear rheology of the CNC-based soft glass and adopting a suitable constitutive model, it is possible to predict the structure-property relationship of the SG under mixed flows. EXPERIMENTS: We use an aqueous suspension with 2 wt% CNC at 25 mM NaCl to form a structurally ordered SG composed of a CNC network containing nematic domains. We combine rheometry and microfluidic experiments with numerical simulations to study the flow properties of the SG in shear, extension, and mixed flow conditions. Extensional flow is investigated in the Optimised Shape Cross-slot Extensional Rheometer (OSCER), where the SG is exposed to shear-free planar elongation. Mixed flow kinematics are investigated in a benchmark microfluidic cylinder device (MCD) where the SG flows past a confined cylinder in a microchannel. FINDINGS: The SG in the MCD displays a velocity overshoot (negative wake) and a pronounced CNC alignment downstream of the cylinder. Simulations using the thixotropic elasto-visco-plastic (TEVP) model yield near quantitative agreement of the velocity profiles in simple and mixed flows and capture the structural fingerprint of the material. Our results provide a comprehensive link between the structural behaviour of a CNC-based SG and its mechanistic properties, laying foundations for the development of functional, built-to-order soft materials.

Size-selective immunofluorescence of Mycobacterium tuberculosis cells by capillary- and viscous forces.

Rapid, low cost screening of tuberculosis requires an effective enrichment method of Mycobacterium tuberculosis (MTB) cells. Currently, microfiltration and centrifugation steps are frequently used for sample preparation, which are cumbersome and time-consuming. In this study, the size-selective capturing mechanism of a microtip-sensor is presented to directly enrich MTB cells from a sample mixture. When a microtip is withdrawn from a spherical suspension in the radial direction, the cells that are concentrated by AC electroosmosis are selectively enriched to the tip due to capillary- and viscous forces. The size-selectivity is characterized by using polystyrene microspheres, which is then applied to size-selective capture of MTB from a sample mixture. Our approach yields a detection limit of 800 cells mL(-1), one of the highest-sensitivity immunosensors to date.

Formation of supramolecular hydrogel microspheres via microfluidics.

Supramolecular hydrogel microspheres are hydrogel particles formed by the self-assembly of hydrogelators in water, through non-covalent interactions. In this paper, we provide a novel strategy to prepare supramolecular hydrogel microspheres with diameters ranging from 15 to 105 microns by using microfluidics. Since the gelation temperature is ca. 64 degrees C, the aqueous solution containing the hydrogelator was initially set at 70 degrees C so the liquid mixture can be pumped into the microfluidic device. The hydrogelator solution then pinches off into uniform micron size droplets at the narrow orifice of the microfluidic device. While traveling downstream in the microchannel, the self-assembly process occurs inside the droplets and the droplets solidify into microsphere gels when the temperature drops to ca. 64 degrees C and below. Optical and scanning electron microscopy (SEM) demonstrate that compact, entangled, round, cage-like aggregates of hydrogelator were formed within the supramolecular hydrogel microsphere, in contrast to loose and less compact aggregates within bulk hydrogel. Thermal analysis (DSC) indicates that supramolecular hydrogel microspheres are more thermally stable and can immobilize more water molecules, owing to the compact entangled three-dimensional network structures. This observation is of particular importance for potential drug delivery and biomaterials applications.

Air Plasma-Enhanced Covalent Functionalization of Poly(methyl methacrylate): High-Throughput Protein Immobilization for Miniaturized Bioassays.

Miniaturized systems, such as integrated microarray and microfluidic devices, are constantly being developed to satisfy the growing demand for sensitive and high-throughput biochemical screening platforms. Owing to its recyclability, and robust mechanical and optical properties, poly(methyl methacrylate) (PMMA) has become the most sought after material for the large-scale fabrication of these platforms. However, the chemical inertness of PMMA entails the use of complex chemical surface treatments for covalent immobilization of proteins. In addition to being hazardous and incompatible for large-scale operations, conventional biofunctionalization strategies pose high risks of compromising the biomolecular conformations, as well as the stability of PMMA. By exploiting radio frequency (RF) air plasma and standard 1-ethyl-3-(3-(dimethylamino)propyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) chemistry in tandem, we demonstrate a simple yet scalable PMMA functionalization strategy for covalent immobilization (chemisorption) of proteins, such as green fluorescent protein (GFP), while preserving the structural integrities of the proteins and PMMA. The surface density of chemisorbed GFP is shown to be highly dependent on the air plasma energy, initial GFP concentration, and buffer pH, where a maximum GFP surface density of 4 × 10-7 mol/m2 is obtained, when chemisorbed on EDC-NHS-activated PMMA exposed to 27 kJ of air plasma, at pH 7.4. Furthermore, antibody-binding studies validate the preserved biofunctionality of the chemisorbed GFP molecules. Finally, the coupled air plasma and EDC-NHS PMMA biofunctionalization strategy is used to fabricate microfluidic antibody assay devices to detect clinically significant concentrations of Chlamydia trachomatis specific antibodies. By coupling our scalable and tailored air plasma-enhanced PMMA biofunctionalization strategy with microfluidics, we elucidate the potential of fabricating sensitive, reproducible, and sustainable high-throughput protein screening systems, without the need for harsh chemicals and complex instrumentation.

Engineering lipid tubules using nano-sized building blocks: the combinatorial self-assembly of vesicles.

Nano-sized lipid vesicles with tailored properties have been used as building blocks to generate lipid tubules between two glass surfaces. The tubules formed not only have defined orientation, width, and length, but they can also grow to be as long as 13 mm under ambient conditions, without externally supplied flow, temperature control, or catalyzing agents. Tubule membrane and its internal aqueous content can be manipulated by controlling the combination of different vesicle's lipid composition and aqueous entrapment. This self-assembly process opens up new pathways for generating complicated and flexible architectures for use in biocompatible molecular and supramolecular engineering. We demonstrated these possibilities by generating tubules encapsulated with siRNA, tubules with multiple branches, and polymerized fluorescent tubules in a single-throughput self-assembly process.

Sensing and Sensibility: Single-Islet-based Quality Control Assay of Cryopreserved Pancreatic Islets with Functionalized Hydrogel Microcapsules.

Despite decades of research and clinical studies of islet transplantations, finding simple yet reliable islet quality assays that correlate accurately with in vivo potency is still a major challenge, especially for real-time and single-islet-based quality assessment. Herein, proof-of-concept studies of a cryopreserved microcapsule-based quality control assays are presented for single islets. Individual rat pancreatic islets and fluorescent oxygen-sensitive dye (FOSD) are encapsulated in alginate hydrogel microcapsules via a microfluidic device. To test the susceptibility of the microcapsules and the FOSD to cryopreservation, the islet microcapsules containing FOSD are cryopreserved and the islet functionalities (adenosine triphosphate, static insulin release measurement, and oxygen consumption rate) are assessed after freezing and thawing steps. The cryopreserved islet capsules with FOSD remain functional after encapsulation and freezing/thawing procedures, validating a simple yet reliable individual-islet-based quality control method for the entire islet processing procedure prior to transplantation. This work also demonstrates that the functionality of cryopreserved islets can be improved by introducing trehalose into the routinely used cryoprotectant dimethyl sulfoxide. The functionalized alginate hydrogel microcapsules with embedded FOSD and optimized cryopreservation protocol presented in this work serve as a versatile islet quality assay and offer tremendous promise for tackling existing challenges in islet transplantation procedures.

Flow-induced immobilization of glucose oxidase in nonionic micellar nanogels for glucose sensing.

A simple microfluidic platform was utilized to immobilize glucose oxidase (GOx) in a nonionic micellar scaffold. The immobilization of GOx was verified by using a combination of cryogenic electron microscopy (cryo-EM), scanning electron microscopy (SEM), and ultraviolet spectroscopy (UV) techniques. Chronoamperometric measurements were conducted on nanogel-GOx scaffolds under different glucose concentrations, exhibiting linear amperometric responses. Without impacting the lifetime and denaturation of GOx, the nonionic nanogel provides a favorable microenvironment for GOx in biological media. This flow-induced immobilization method in a nonionic nanogel host matrix opens up new pathways for designing a simple, fast, biocompatible, and cost-effective process to immobilize biomolecules that are averse to ionic environments.

Microfluidic one-step fabrication of radiopaque alginate microgels with in situ synthesized barium sulfate nanoparticles.

In this work, we report a new strategy to fabricate monodispersed radiopaque alginate (Ba-alginate) microgels by a one-step microfluidic method. Alginate droplets containing sulfate ions are first formed by a flow focusing microfluidic setup. These alginate droplets are subsequently solidified by barium ions in a collection bath. During the solidification process, excessive barium ions in the collection bath also react with sulfate ions in the alginate droplet, resulting in barium sulfate (BaSO(4)) nanoparticles in situ synthesized (acting as radiopaque imaging agents) within the Ba-alginate microgels. Scanning electron microscopy (SEM) combined with energy-dispersive X-ray spectroscopy (EDX) illustrate that 800 nm BaSO(4) nanoparticles are uniformly distributed inside the 30 µm Ba-alginate microgels, with 62 wt% of elemental barium (Ba). In addition, X-ray diffraction (XRD) measurements indicate that the BaSO(4) nanoparticles consist of 10 nm in situ synthesized BaSO(4) crystallites. The alginate microgels act as a soft and porous template to prevent the precipitation and aggregation of BaSO(4) nanoparticles. The Ba-alginate microgels are also visible under X-ray radiation. The facile route to fabricate alginate microgels as radiopaque embolic materials is of particular importance for endovascular embolization and localized diagnostic imaging applications. Similar approaches can also be adopted for synthesizing other inorganic nanoparticles in microgels.