Direct Fabrication of Functional Shapes on 3D Surfaces Using Electrospinning.

In this work, we demonstrate the ability to simultaneously pattern fibers and fabricate functional 2D and 3D shapes (e.g., letters, mask-like structures with nose bridges and ear loops, aprons, hoods) using a single step electrospinning process. Using 2D and 3D mesh templates, electrospun fibers were preferentially attracted to the metal protrusions relative to the voids so that the pattern of the electrospun mat mimicked the woven mesh macroscopically. On a microscopic scale, the electrostatic lensing effect decreased fiber diameter and narrowed the fiber size distribution, e.g., the coefficient of variation of the fiber diameter for sample collected on a 0.6 mm mesh was 14% compared to 55% for the sample collected on foil). Functionally, the mesh did not affect the wettability of the fiber mats. Notably, the fiber patterning increased the rigidity of the fiber mat. There was a 2-fold increase in flexural rigidity using the 0.6 mm mesh compared to the sample collected on foil. Overall, we anticipate this approach will be a versatile tool for design and fabrication of 2D and 3D patterns with potential applications in personalized wound care and surgical meshes.

Metabolic Heterogeneity of Cerebral Cortical and Cerebellar Astrocytes.

Astrocytes play critical roles in regulating neuronal synaptogenesis, maintaining blood-brain barrier integrity, and recycling neurotransmitters. Increasing numbers of studies have suggested astrocyte heterogeneity in morphology, gene profile, and function. However, metabolic phenotype of astrocytes in different brain regions have not been explored. In this paper, we investigated the metabolic signature of cortical and cerebellar astrocytes using primary astrocyte cultures. We observed that cortical astrocytes were larger than cerebellar astrocytes, whereas cerebellar astrocytes had more and longer processes than cortical astrocytes. Using a Seahorse extracellular flux analyzer, we demonstrated that cortical astrocytes had higher mitochondrial respiration and glycolysis than cerebellar astrocytes. Cerebellar astrocytes have lower spare capacity of mitochondrial respiration and glycolysis as compared with cortical astrocytes. Consistently, cortical astrocytes have higher mitochondrial oxidation and glycolysis-derived ATP content than cerebellar astrocytes. In addition, cerebellar astrocytes have a fuel preference for glutamine and fatty acid, whereas cortical astrocytes were more dependent on glucose to meet energy demands. Our study indicated that cortical and cerebellar astrocytes display distinct metabolic phenotypes. Future studies on astrocyte metabolic heterogeneity and brain function in aging and neurodegeneration may lead to better understanding of the role of astrocyte in brain aging and neurodegenerative disorders.

Recurrent Transient Ischemic Attack Induces Neural Cytoskeleton Modification and Gliosis in an Experimental Model.

Transient ischemic attack (TIA) presents a high risk for subsequent stroke, Alzheimer's disease (AD), and related dementia (ADRD). However, the neuropathophysiology of TIA has been rarely studied. By evaluating recurrent TIA-induced neuropathological changes, our study aimed to explore the potential mechanisms underlying the contribution of TIA to ADRD. In the current study, we established a recurrent TIA model by three times 10-min middle cerebral artery occlusion within a week in rat. Neither permanent neurological deficit nor apoptosis was observed following recurrent TIA. No increase of AD-related biomarkers was indicated after TIA, including increase of tau hyperphosphorylation and ß-site APP cleaving enzyme 1 (BACE1). Neuronal cytoskeleton modification and neuroinflammation was found at 1, 3, and 7 days after recurrent TIA, evidenced by the reduction of microtubule-associated protein 2 (MAP2), elevation of neurofilament-light chain (NFL), and increase of glial fibrillary acidic protein (GFAP)-positive astrocytes and ionized calcium binding adaptor molecule 1 (Iba1)-positive microglia at the TIA-affected cerebral cortex and basal ganglion. Similar NFL, GFAP and Iba1 alteration was found in the white matter of corpus callosum. In summary, the current study demonstrated that recurrent TIA may trigger neuronal cytoskeleton change, astrogliosis, and microgliosis without induction of cell death at the acute and subacute stage. Our study indicates that TIA-induced neuronal cytoskeleton modification and neuroinflammation may be involved in the vascular contribution to cognitive impairment and dementia.

Cascade Processes with Micellar Reaction Media: Recent Advances and Future Directions.

Reducing the use of solvents is an important aim of green chemistry. Using micelles self-assembled from amphiphilic molecules dispersed in water (considered a green solvent) has facilitated reactions of organic compounds. When performing reactions in micelles, the hydrophobic effect can considerably accelerate apparent reaction rates, as well as enhance selectivity. Here, we review micellar reaction media and their potential role in sustainable chemical production. The focus of this review is applications of engineered amphiphilic systems for reactions (surface-active ionic liquids, designer surfactants, and block copolymers) as reaction media. Micelles are a versatile platform for performing a large array of organic chemistries using water as the bulk solvent. Building on this foundation, synthetic sequences combining several reaction steps in one pot have been developed. Telescoping multiple reactions can reduce solvent waste by limiting the volume of solvents, as well as eliminating purification processes. Thus, in particular, we review recent advances in "one-pot" multistep reactions achieved using micellar reaction media with potential applications in medicinal chemistry and agrochemistry. Photocatalyzed reactions in micellar reaction media are also discussed. In addition to the use of micelles, we emphasize the process (steps to isolate the product and reuse the catalyst).

Polymer Nanoparticles Enhance Irreversible Electroporation In Vitro.

Expanding the volume of an irreversible electroporation treatment typically necessitates an increase in pulse voltage, number, duration, or repetition. This study investigates the addition of polyethylenimine nanoparticles (PEI-NP) to pulsed electric field treatments, determining their combined effect on ablation size and voltages. U118 cells in an in vitro 3D cell culture model were treated with one of three pulse parameters (with and without PEI-NPs) which are representative of irreversible electroporation (IRE), high frequency irreversible electroporation (H-FIRE), or nanosecond pulsed electric fields (nsPEF). The size of the ablations were compared and mapped onto an electric field model to describe the electric field required to induce cell death. Analysis was conducted to determine the role of PEI-NPs in altering media conductivity, the potential for PEI-NP degradation following pulsed electric field treatment, and PEI-NP uptake. Results show there was a statistically significant increase in ablation diameter for IRE and H-FIRE pulses with PEI-NPs. There was no increase in ablation size for nsPEF with PEI-NPs. This all occurs with no change in cell media conductivity, no observable degradation of PEI-NPs, and moderate particle uptake. These results demonstrate the synergy of a combined cationic polymer nanoparticle and pulsed electric field treatment for the ablation of cancer cells. These results set the foundation for polymer nanoparticles engineered specifically for irreversible electroporation.

Amphiphilic Polymer Nanoreactors for Multiple Step, One-Pot Reactions and Spontaneous Product Separation.

Performing multiple reaction steps in "one pot" to avoid the need to isolate intermediates is a promising approach for reducing solvent waste associated with liquid phase chemical processing. In this work, we incorporated gold nanoparticle catalysts into polymer nanoreactors via amphiphilic block copolymer directed self-assembly. With the polymer nanoreactors dispersed in water as the bulk solvent, we demonstrated the ability to facilitate two reaction steps in one pot with spontaneous precipitation of the product from the reaction mixture. Specifically, we achieved imide synthesis from 4-nitrophenol and benzaldehyde as a model reaction. The reaction occured in water at ambient conditions; the desired 4-benzylideneaminophenol product spontaneously precipitated from the reaction mixture while the nanoreactors remained stable in dispersion. A 65% isolated yield was achieved. In contrast, PEGylated gold nanoparticles and citrate stabilized gold nanoparticles precipitated with the reaction product, which would complicate both the isolation of the product as well as reuse of the catalyst. Thus, amphiphilic nanoreactors dispersed in water are a promising approach for reducing solvent waste associated with liquid phase chemical processing by using water as the bulk solvent, eliminating the need to isolate intermediates, achieving spontaneous product separation to facilitate the recycling of the reaction mixture, and simplifying the isolation of the desired product.

Identifying Chemical Reactions and Their Associated Attributes in Patents.

Chemical patents are an essential source of information about novel chemicals and chemical reactions. However, with the increasing volume of such patents, mining information about these chemicals and chemical reactions has become a time-intensive and laborious endeavor. In this study, we present a system to extract chemical reaction events from patents automatically. Our approach consists of two steps: 1) named entity recognition (NER)-the automatic identification of chemical reaction parameters from the corresponding text, and 2) event extraction (EE)-the automatic classifying and linking of entities based on their relationships to each other. For our NER system, we evaluate bidirectional long short-term memory (BiLSTM)-based and bidirectional encoder representations from transformer (BERT)-based methods. For our EE system, we evaluate BERT-based, convolutional neural network (CNN)-based, and rule-based methods. We evaluate our NER and EE components independently and as an end-to-end system, reporting the precision, recall, and F 1 score. Our results show that the BiLSTM-based method performed best at identifying the entities, and the CNN-based method performed best at extracting events.

Polymeric Nanoparticle Delivery of Combination Therapy with Synergistic Effects in Ovarian Cancer.

Treatment of ovarian cancer is challenging due to late stage diagnosis, acquired drug resistance mechanisms, and systemic toxicity of chemotherapeutic agents. Combination chemotherapy has the potential to enhance treatment efficacy by activation of multiple downstream pathways to overcome drug resistance and reducing required dosages. Sequence of delivery and the dosing schedule can further enhance treatment efficacy. Formulation of drug combinations into nanoparticles can further enhance treatment efficacy. Due to their versatility, polymer-based nanoparticles are an especially promising tool for clinical translation of combination therapies with tunable dosing schedules. We review polymer nanoparticle (e.g., micelles, dendrimers, and lipid nanoparticles) carriers of drug combinations formulated to treat ovarian cancer. In particular, the focus on this review is combinations of platinum and taxane agents (commonly used first line treatments for ovarian cancer) combined with other small molecule therapeutic agents. In vitro and in vivo drug potency are discussed with a focus on quantifiable synergistic effects. The effect of drug sequence and dosing schedule is examined. Computational approaches as a tool to predict synergistic drug combinations and dosing schedules as a tool for future nanoparticle design are also briefly discussed.

Rheological characterization of poly-dimethyl siloxane formulations with tunable viscoelastic properties.

Studies from the past two decades have demonstrated convincingly that cells are able to sense the mechanical properties of their surroundings. Cells make major decisions in response to this mechanosensation, including decisions regarding cell migration, proliferation, survival, and differentiation. The vast majority of these studies have focused on the cellular mechanoresponse to changing substrate stiffness (or elastic modulus) and have been conducted on purely elastic substrates. In contrast, most soft tissues in the human body exhibit viscoelastic behavior; that is, they generate responsive force proportional to both the magnitude and rate of strain. While several recent studies have demonstrated that viscous effects of an underlying substrate affect cellular mechanoresponse, there is not a straightforward experimental method to probe this, particularly for investigators with little background in biomaterial fabrication. In the current work, we demonstrate that polymers comprised of differing polydimethylsiloxane (PDMS) formulations can be generated that allow for control over both the strain-dependent storage modulus and the strain rate-dependent loss modulus. These substrates requires no background in biomaterial fabrication to fabricate, are shelf-stable, and exhibit repeatable mechanical properties. Here we demonstrate that these substrates are biocompatible and exhibit similar protein adsorption characteristics regardless of mechanical properties. Finally, we develop a set of empirical equations that predicts the storage and loss modulus for a given blend of PDMS formulations, allowing users to tailor substrate mechanical properties to their specific needs.

Self-Assembly of pH-Labile Polymer Nanoparticles for Paclitaxel Prodrug Delivery: Formulation, Characterization, and Evaluation.

The efficacy of paclitaxel (PTX) is limited due to its poor solubility, poor bioavailability, and acquired drug resistance mechanisms. Designing paclitaxel prodrugs can improve its anticancer activity and enable formulation of nanoparticles. Overall, the aim of this work is to improve the potency of paclitaxel with prodrug synthesis, nanoparticle formation, and synergistic formulation with lapatinib. Specifically, we improve potency of paclitaxel by conjugating it to α-tocopherol (vitamin E) to produce a hydrophobic prodrug (Pro); this increase in potency is indicated by the 8-fold decrease in half maximal inhibitory concentration (IC50) concentration in ovarian cancer cell line, OVCA-432, used as a model system. The efficacy of the paclitaxel prodrug was further enhanced by encapsulation into pH-labile nanoparticles using Flash NanoPrecipitation (FNP), a rapid, polymer directed self-assembly method. There was an 1100-fold decrease in IC50 concentration upon formulating the prodrug into nanoparticles. Notably, the prodrug formulations were 5-fold more potent than paclitaxel nanoparticles. Finally, the cytotoxic effects were further enhanced by co-encapsulating the prodrug with lapatinib (LAP). Formulating the drug combination resulted in synergistic interactions as indicated by the combination index (CI) of 0.51. Overall, these results demonstrate this prodrug combined with nanoparticle formulation and combination therapy is a promising approach for enhancing paclitaxel potency.

Accelerated Reaction Rates within Self-Assembled Polymer Nanoreactors with Tunable Hydrophobic Microenvironments.

Performing reactions in the presence of self-assembled hierarchical structures of amphiphilic macromolecules can accelerate reactions while using water as the bulk solvent due to the hydrophobic effect. We leveraged non-covalent interactions to self-assemble filled-polymer micelle nanoreactors (NR) incorporating gold nanoparticle catalysts into various amphiphilic polymer nanostructures with comparable hydrodynamic nanoreactor size and gold concentration in the nanoreactor dispersion. We systematically studied the effect of the hydrophobic co-precipitant on self-assembly and catalytic performance. We observed that co-precipitants that interact with gold are beneficial for improving incorporation efficiency of the gold nanoparticles into the nanocomposite nanoreactor during self-assembly but decrease catalytic performance. Hierarchical assemblies with co-precipitants that leverage noncovalent interactions could enhance catalytic performance. For the co-precipitants that do not interact strongly with gold, the catalytic performance was strongly affected by the hydrophobic microenvironment of the co-precipitant. Specifically, the apparent reaction rate per surface area using castor oil (CO) was over 8-fold greater than polystyrene (750 g/mol, PS 750); the turnover frequency was higher than previously reported self-assembled polymer systems. The increase in apparent catalytic performance could be attributed to differences in reactant solubility rather than differences in mass transfer or intrinsic kinetics; higher reactant solubility enhances apparent reaction rates. Full conversion of 4-nitrophenol was achieved within three minutes for at least 10 sequential reactions demonstrating that the nanoreactors could be used for multiple reactions.

Color Space Transformation-Based Algorithm for Evaluation of Thermochromic Behavior of Cholesteric Liquid Crystals Using Polarized Light Microscopy.

Cholesteryl ester liquid crystals exhibit thermochromic properties related to the existence of a twisted nematic phase. When used in applications such as thermal mapping, a color change is often monitored by video cameras. Thus, quantitative methods to evaluate thermochromic behavior (e.g., blue-start, red-start, red-end, color play and bandwidth) from video analysis are desirable. However, obtaining quantitative color measurements from digital images remains a significant technical challenge, especially for highly reflective samples such as liquid crystals (for which ultraviolet-visible (UV-vis) reflectance spectroscopy is typically used). We developed a method to determine thermochromic properties from videos of liquid crystal cooling under polarized light microscopy. We relate observed color transitions to quantifiable changes in the cumulative color difference in the International Commission on Illumination (CIE) L*a*b* color space and validate this method with UV-vis reflectance spectroscopy. The measured thermochromic behavior and associated measurement uncertainties (coefficient of variations) were comparable to UV-vis reflectance measurements.

Thermochromic Fibers via Electrospinning.

Cholesteryl ester liquid crystals exhibit thermochromic properties related to the existence of a twisted nematic phase. We formulate ternary mixtures of cholesteryl benzoate (CB), cholesteryl pelargonate (CP), and cholesteryl oleyl carbonate (COC) to achieve thermochromic behavior. We aim to achieve thermochromic fibers by incorporating the liquid crystal formulations into electrospun fibers. Two methods of incorporating the liquid crystal (LC) are compared: (1) blend electrospinning and (2) coaxial electrospinning using the same solvent system for the liquid crystal. For blend electrospinning, intermolecular interactions seem to be important in facilitating fiber formation since addition of LC can suppress bead formation. Coaxial electrospinning produces fibers with higher nominal fiber production rates (g/hr) and with higher nominal LC content in the fiber (wt. LC/wt. polymer assuming all of the solvent evaporates) but larger fiber size distributions as quantified by the coefficient of variation in fiber diameter than blend electrospinning with a single nozzle. Importantly, our proof-of-concept experiments demonstrate that coaxially electrospinning with LC and solvent in the core preserves the thermochromic properties of the LC so that thermochromic fibers are achieved.

Rapid Self-Assembly of Polymer Nanoparticles for Synergistic Codelivery of Paclitaxel and Lapatinib via Flash NanoPrecipitation.

Taxol, a formulation of paclitaxel (PTX), is one of the most widely used anticancer drugs, particularly for treating recurring ovarian carcinomas following surgery. Clinically, PTX is used in combination with other drugs such as lapatinib (LAP) to increase treatment efficacy. Delivering drug combinations with nanoparticles has the potential to improve chemotherapy outcomes. In this study, we use Flash NanoPrecipitation, a rapid, scalable process to encapsulate weakly hydrophobic drugs (logP < 6) PTX and LAP into polymer nanoparticles with a coordination complex of tannic acid and iron formed during the mixing process. We determine the formulation parameters required to achieve uniform nanoparticles and evaluate the drug release in vitro. The size of the resulting nanoparticles was stable at pH 7.4, facilitating sustained drug release via first-order Fickian diffusion. Encapsulating either PTX or LAP into nanoparticles increases drug potency (as indicated by the decrease in IC-50 concentration); we observe a 1500-fold increase in PTX potency and a six-fold increase in LAP potency. When PTX and LAP are co-loaded in the same nanoparticle, they have a synergistic effect that is greater than treating with two single-drug-loaded nanoparticles as the combination index is 0.23 compared to 0.40, respectively.

Rapid, Single-Step Protein Encapsulation via Flash NanoPrecipitation.

Flash NanoPrecipitation (FNP) is a rapid method for encapsulating hydrophobic materials in polymer nanoparticles with high loading capacity. Encapsulating biologics such as proteins remains a challenge due to their low hydrophobicity (logP < 6) and current methods require multiple processing steps. In this work, we report rapid, single-step protein encapsulation via FNP using bovine serum albumin (BSA) as a model protein. Nanoparticle formation involves complexation and precipitation of protein with tannic acid and stabilization with a cationic polyelectrolyte. Nanoparticle self-assembly is driven by hydrogen bonding and electrostatic interactions. Using this approach, high encapsulation efficiency (up to ~80%) of protein can be achieved. The resulting nanoparticles are stable at physiological pH and ionic strength. Overall, FNP is a rapid, efficient platform for encapsulating proteins for various applications.

Single-Step Self-Assembly and Physical Crosslinking of PEGylated Chitosan Nanoparticles by Tannic Acid.

Chitosan-based nanoparticles are promising materials for potential biomedical applications. We used Flash NanoPrecipitation as a rapid, scalable, single-step method to achieve self-assembly of crosslinked chitosan nanoparticles. Self-assembly was driven by electrostatic interactions, hydrogen bonding, and hydrophobic interactions; tannic acid served to precipitate chitosan to seed nanoparticle formation and crosslink the chitosan to stabilize the resulting particles. The size of the nanoparticles can be tuned by varying formulation parameters including the total solids concentration and block copolymer to core mass ratio. We demonstrated that hydrophobic moieties can be incorporated into the nanoparticle using a lipophilic fluorescent dye as a model system.

Shear Force Fiber Spinning: Process Parameter and Polymer Solution Property Considerations.

For application of polymer nanofibers (e.g., sensors, and scaffolds to study cell behavior) it is important to control the spatial orientation of the fibers. We compare the ability to align and pattern fibers using shear force fiber spinning, i.e. contacting a drop of polymer solution with a rotating collector to mechanically draw a fiber, with electrospinning onto a rotating drum. Using polystyrene as a model system, we observe that the fiber spacing using shear force fiber spinning was more uniform than electrospinning with the rotating drum with relative standard deviations of 18% and 39%, respectively. Importantly, the approaches are complementary as the fiber spacing achieved using electrospinning with the rotating drum was ~10 microns while fiber spacing achieved using shear force fiber spinning was ~250 microns. To expand to additional polymer systems, we use polymer entanglement and capillary number. Solution properties that favor large capillary numbers (>50) prevent droplet breakup to facilitate fiber formation. Draw-down ratio was useful for determining appropriate process conditions (flow rate, rotational speed of the collector) to achieve continuous formation of fibers. These rules of thumb for considering the polymer solution properties and process parameters are expected to expand use of this platform for creating hierarchical structures of multiple fiber layers for cell scaffolds and additional applications.

Rapid Self-Assembly of Metal/Polymer Nanocomposite Particles as Nanoreactors and Their Kinetic Characterization.

Self-assembled metal nanoparticle-polymer nanocomposite particles as nanoreactors are a promising approach for performing liquid phase reactions using water as a bulk solvent. In this work, we demonstrate rapid, scalable self-assembly of metal nanoparticle catalyst-polymer nanocomposite particles via Flash NanoPrecipitation. The catalyst loading and size of the nanocomposite particles can be tuned independently. Using nanocomposite particles as nanoreactors and the reduction of 4-nitrophenol as a model reaction, we study the fundamental interplay of reaction and diffusion. The induction time is affected by the sequence of reagent addition, time between additions, and reagent concentration. Combined, our experiments indicate the induction time is most influenced by diffusion of sodium borohydride. Following the induction time, scaling analysis and effective diffusivity measured using NMR indicate that the observed reaction rate are reaction- rather than diffusion-limited. Furthermore, the intrinsic kinetics are comparable to ligand-free gold nanoparticles. This result indicates that the polymer microenvironment does not de-activate or block the catalyst active sites.

Cystic Fibrosis Sputum Rheology Correlates With Both Acute and Longitudinal Changes in Lung Function.

BACKGROUND: Cystic fibrosis (CF) airway secretions are abnormal, contributing to decreased clearance and a cycle of infection and inflammation. CF sputum properties may predict disease progression. We hypothesized that sputum viscoelasticity and clearance abnormalities would inversely correlate with pulmonary function during exacerbation and that sputum properties would return to baseline after therapy. METHODS: We collected sputa longitudinally from 13 subjects with CF with moderate to severe lung disease during both clinical stability and exacerbation. Dynamic rheology was analyzed, using a cone-and-plate rheometer. Mucociliary clearability was measured on mucus-depleted frog palate, cough clearability in a simulated cough machine, and sputum hydration as percent solids was measured following lyophilization. RESULTS: Elastic modulus G' and viscous modulus G'' increased during exacerbation and returned to baseline levels with recovery (P < .05 for both). Solid content did not change. Sputum mucociliary clearability decreased during exacerbations (P < .01) but not cough clearance. FEV1 % predicted was inversely correlated with G' and G'' (P < .01 for both). The regression slope of the natural log-transformed G' and G'' vs FEV1 % predicted was statistically homogeneous among subjects (estimated common slope m = -3.84, P < .001 and m = -8.53, P < .0001, respectively). CONCLUSIONS: Among these subjects with CF, there is a striking identity of the slope defining the relationship between ln G' or ln G'' and FEV1. There are dramatic increases in dynamic viscosity and elasticity during a pulmonary exacerbation with return to baseline at recovery. This suggests that sputum viscoelastic properties are tightly associated with lung function and disease status.

Controlling and Predicting Nanoparticle Formation by Block Copolymer Directed Rapid Precipitations.

Nanoparticles have shown promise in several biomedical applications, including drug delivery and medical imaging; however, quantitative prediction of nanoparticle formation processes that scale from laboratory to commercial production has been lacking. Flash NanoPrecipitation (FNP) is a scalable technique to form highly loaded, block copolymer protected nanoparticles. Here, the FNP process is shown to strictly obey diffusion-limited aggregation assembly kinetics, and the parameters that control the nanoparticle size and the polymer brush density on the nanoparticle surface are shown. The particle size, ranging from 40 to 200 nm, is insensitive to the molecular weight and chemical composition of the hydrophobic encapsulated material, which is shown to be a consequence of the diffusion-limited growth kinetics. In a simple model derived from these kinetics, a single constant describes the 46 unique nanoparticle formulations produced here. The polymer brush densities on the nanoparticle surface are weakly dependent on the process parameters and are among the densest reported in the literature. Though modest differences in brush densities are observed, there is no measurable difference in the amount of protein adsorbed within this range. This work highlights the material-independent and universal nature of the Flash NanoPrecipitation process, allowing for the rapid translation of formulations to different stabilizing polymers and therapeutic loads.