Sculpting liquid metal stabilized interfaces: a gateway to liquid electronics.

Liquid electronics have potential applications in soft robotics, printed electronics, and healable electronics. The intrinsic shortcomings of solid-state electronics can be offset by liquid conductors. Alloys of gallium have emerged as transformative materials for liquid electronics owing to their intrinsic fluidity, conductivity, and low toxicity. However, sculpting liquid metal or its composites into a 3D architecture is a challenging task. To tackle this issue, herein, we explored the interfacial chemistry of metal ions and tannic acid (TA) complexation at a liquid-liquid interface. First, we established that an MIII-TA network at the liquid-liquid interface could structure liquid in liquid by jamming the interfacial film. The surface coverage of the droplet largely depends on the concentration of metal ions, oxidation state of metal ions and pH of the surrounding environment. Further extending the approach, we demonstrated that TA-functionalized gallium nanoparticles (Ga NPs) can also sculpt liquid droplets in the presence of transition metal ions. Finally, a mold-based free-standing 3D architecture is obtained using the interfacial reaction and interfacial crowding of a metal-phenolate network. Conductivity measurement reveals that these liquid constructs can be used for low-voltage electronic applications, thus opening the door for liquid electronics.

Identification of a myofibroblast differentiation program during neonatal lung development.

Alveologenesis is the final stage of lung development in which the internal surface area of the lung is increased to facilitate efficient gas exchange in the mature organism. The first phase of alveologenesis involves the formation of septal ridges (secondary septae) and the second phase involves thinning of the alveolar septa. Within secondary septa, mesenchymal cells include a transient population of alveolar myofibroblasts (MyoFBs) and a stable but poorly described population of lipid-rich cells that have been referred to as lipofibroblasts or matrix fibroblasts (MatFBs). Using a unique Fgf18CreER lineage trace mouse line, cell sorting, single-cell RNA sequencing and primary cell culture, we have identified multiple subtypes of mesenchymal cells in the neonatal lung, including an immature progenitor cell that gives rise to mature MyoFB. We also show that the endogenous and targeted ROSA26 locus serves as a sensitive reporter for MyoFB maturation. These studies identify a MyoFB differentiation program that is distinct from other mesenchymal cell types and increases the known repertoire of mesenchymal cell types in the neonatal lung.

Supramolecular Modulation of Fluid Flow in a Self-Powered Enzyme Micropump.

Regulating macroscopic fluid flow by catalytic harnessing of chemical energy could potentially provide a solution for powerless microfluidic devices. Earlier reports have shown that surface-anchored enzymes can actuate the surrounding fluid in the presence of the respective substrate in a concentration-dependent manner. It is also crucial to have control over the flow speed of a self-powered enzyme micropump in various applications where controlled dosing and mixing are required. However, modulating the flow speed independent of the fuel concentration remains a significant challenge. In a quest to regulate the fluid flow in such a system, a supramolecular approach has been adopted, where reversible regulation of enzyme activity was achieved by a two-faced synthetic receptor bearing sulfonamide and adamantane groups. The bovine carbonic anhydrase (BCA) enzyme containing a single binding site favorable to the sulfonamide group was used as a model enzyme, and the enzyme activity was inhibited in the presence of the two-faced inhibitor. The same effect was reflected when the immobilized enzyme was used as an engine to actuate the fluid flow. The flow velocity was reduced up to 53% in the presence of 100 µM inhibitor. Later, upon addition of a supramolecular "host" CB[7], the inhibitor was sequestered from the enzyme due to the higher binding affinity of CB[7] with the adamantane functionality of the inhibitor. As a result, the flow velocity was restored to ∼72%, thus providing successful supramolecular control over a self-powered enzyme micropump.

Early Resveratrol Treatment Mitigates Joint Degeneration and Dampens Pain in a Mouse Model of Pseudoachondroplasia (PSACH).

Pseudoachondroplasia (PSACH), a severe dwarfing condition associated with early-onset joint degeneration and lifelong joint pain, is caused by mutations in cartilage oligomeric matrix protein (COMP). The mechanisms underlying the mutant-COMP pathology have been defined using the MT-COMP mouse model of PSACH that has the common D469del mutation. Mutant-COMP protein does not fold properly, and it is retained in the rough endoplasmic reticulum (rER) of chondrocytes rather than being exported to the extracellular matrix (ECM), driving ER stress that stimulates oxidative stress and inflammation, driving a self-perpetuating cycle. CHOP (ER stress signaling protein) and TNFα inflammation drive high levels of mTORC1 signaling, shutting down autophagy and blocking ER clearance, resulting in premature loss of chondrocytes that negatively impacts linear growth and causes early joint degeneration in MT-COMP mice and PSACH. Previously, we have shown that resveratrol treatment from birth to 20 weeks prevents joint degeneration and decreases the pathological processes in articular chondrocytes. Resveratrol's therapeutic mechanism of action in the mutant-COMP pathology was shown to act by primarily stimulating autophagy and reducing inflammation. Importantly, we demonstrated that MT-COMP mice experience pain consistent with PSACH joint pain. Here, we show, in the MT-COMP mouse, that resveratrol treatment must begin within 4 weeks to preserve joint health and reduce pain. Resveratrol treatment started at 6 or 8 weeks (to 20 weeks) was not effective in preventing joint degeneration. Collectively, our findings in MT-COMP mice show that there is a postnatal resveratrol treatment window wherein the inevitable mutant-COMP joint degeneration and pain can be prevented.

Autonomous macroscopic signal deciphering the geometric self-sorting of pillar[n]arenes.

In this communication, we have deciphered the geometric self-sorting of pillar[n]arenes by analyzing the fluid flow pattern obtained during the self-assembly of complementary pillar[n]arenes on the surface. The concept was further extended to demonstrate flow manipulation inside a microchannel where multiple sites were available for self-sorting, and the resultant flow velocity was tuned by the feeding ratio of the complementary pairs.

Targeted deletion of Fgf9 in tendon disrupts mineralization of the developing enthesis.

The enthesis is a transitional tissue between tendon and bone that matures postnatally. The development and maturation of the enthesis involve cellular processes likened to an arrested growth plate. In this study, we explored the role of fibroblast growth factor 9 (Fgf9), a known regulator of chondrogenesis and vascularization during bone development, on the structure and function of the postnatal enthesis. First, we confirmed spatial expression of Fgf9 in the tendon and enthesis using in situ hybridization. We then used Cre-lox recombinase to conditionally knockout Fgf9 in mouse tendon and enthesis (Scx-Cre) and characterized enthesis morphology as well as mechanical properties in Fgf9ScxCre and wild-type (WT) entheses. Fgf9ScxCre mice had smaller calcaneal and humeral apophyses, thinner cortical bone at the attachment, increased cellularity, and reduced failure load in mature entheses compared to WT littermates. During postnatal development, we found reduced chondrocyte hypertrophy and disrupted type X collagen (Col X) in Fgf9ScxCre entheses. These findings support that tendon-derived Fgf9 is important for functional development of the enthesis, including its postnatal mineralization. Our findings suggest the potential role of FGF signaling during enthesis development.

Discrimination of enantiomers and constitutional isomers by self-generated macroscopic fluid flow.

The amplification of weak molecular signals to visible output could provide a gateway to the macroscopic world. In this context, supramolecular interfaces were designed by depositing macrocyclic "host" molecules in a multilayer film that can be utilized to discriminate isomers by their fluid flow response upon "host-guest" molecular recognition.

Identification of a myofibroblast differentiation program during neonatal lung development.

Alveologenesis is the final stage of lung development in which the internal surface area of the lung is increased to facilitate efficient gas exchange in the mature organism. The first phase of alveologenesis involves the formation of septal ridges (secondary septae) and the second phase involves thinning of the alveolar septa. Within secondary septa, mesenchymal cells include a transient population of alveolar myofibroblasts (MyoFB) and a stable but poorly described population of lipid rich cells that have been referred to as lipofibroblasts or matrix fibroblasts (MatFB). Using a unique Fgf18CreER lineage trace mouse line, cell sorting, single cell RNA sequencing, and primary cell culture, we have identified multiple subtypes of mesenchymal cells in the neonatal lung, including an immature progenitor cell that gives rise to mature MyoFB. We also show that the endogenous and targeted ROSA26 locus serves as a sensitive reporter for MyoFB maturation. These studies identify a myofibroblast differentiation program that is distinct form other mesenchymal cells types and increases the known repertoire of mesenchymal cell types in the neonatal lung.

A visible-light activated ROS generator multilayer film for antibacterial coatings.

The current scenario of antibiotic-resistant bacteria and pandemics caused by viruses makes research in the area of antibacterial and antiviral materials and surfaces more urgent than ever. In this regard, salicylideneimine based tetracoordinate boron-containing organic compounds are emerging as a new class of photosensitizers for singlet oxygen generation. However, the inherent inability of small organic molecules to be processed limits their potential use in functional coatings. Here we show the synthesis of a novel polymer functionalized with diiodosalicylideneimine-boron difluoride (PEI-BF2) and its utility for surface coating inside glass vials via layer-by-layer (LbL) assembly. The multilayer thin films are characterized using AFM and UV-Vis spectroscopy and the resultant coatings display excellent stability. The multilayer coating could be activated using visible light, and owing to the photocatalytic activity of the incorporated PEI-BF2, the surface coating is able to generate singlet oxygen efficiently upon light irradiation. Further, the multilayer coated surfaces exhibit remarkable antimicrobial activity towards both Gram-positive and Gram-negative bacteria under a variety of conditions. Thus, owing to the simple synthesis and the convenient methodology adopted for the preparation of multilayer coatings, the material reported here could pave the way for the development of sunlight activated large area self-sterile surfaces.

Polymer multilayer films regulate macroscopic fluid flow and power microfluidic devices via supramolecular interactions.

Self-powered supramolecular micropumps could potentially provide a solution for powerless microfluidic devices where the fluid flow can be manipulated via modulating non-covalent interactions. An attempt has been made to fabricate thin-film-based micropumps by depositing a ß-cyclodextrin ('host') functionalized polymer on a glass slide via layer-by-layer assembly. These supramolecular micropumps turned on the fluid flow upon addition of 'guest' molecules to the multilayer films. The flow velocity was tuned using the concentration of the guest molecules as well as the number of host layers inside the multilayer films. Numerical modelling reveals that the solutal buoyancy, which originates from host-guest complexation, is primarily responsible for the fluid flow. In view of its potential application in self-powered devices, the thin-film-based micropump was integrated into a microfluidic device to show molecular and colloidal transport over long distances.

Anisotropic Compartmentalization of the Liquid-Liquid Interface using Dynamic Imine Chemistry.

The liquid-liquid interface offers a fascinating avenue for generating hierarchical compartments. Herein, the dynamic imine chemistry is employed at the oil-water interface to investigate the effect of dynamic covalent bonds for modulating the droplet shape. The imine bond formation between oil-soluble aromatic aldehydes and water-soluble polyethyleneimine greatly stabilized the oil-water interface by substantially lowering the interfacial tension. The successful jamming of imine-mediated assemblies was observed when a compressive force was applied to the droplet. Thus, the anisotropic compartmentalization of the liquid-liquid interface was created, and it was later altered by changing the pH of the surrounding environment. Finally, a proof-of-concept demonstration of a pH-triggered cargo release across the interfacial membrane confirmed the feasibility of stimuli-responsive behavior of dynamic imine assemblies.

Inhibitory effect of nucleotides on acetylcholine esterase activity and its microflow-based actuation in blood plasma.

The inhibitory effect of nucleotides on the catalytic activity of acetylcholine esterase (AChE) was rationalized and a similar inhibition trend was observed when analyzing the macroscopic fluid flow generated by surface immobilized AChE. Additionally, the demonstration of enzymatic micropumping by showing adenine-nucleotide responsive AChE actuated fluid flow from blood plasma paved the way for designing future lab-on-a-chip devices in complex biological environments with potential clinical applications.

Fluid actuation and buoyancy driven oscillation by enzyme-immobilized microfluidic microcapsules.

Mimicking microorganism's locomotion and actuation under fluid is difficult to realize. To better comprehend the motility in non-living matter, self-propelled synthetic systems are being developed as a fast-growing area of research. Inspired by the self-powered enzyme micropumps where the enzyme catalysis was harnessed to create motion, herein, enzyme-immobilized microfluidic microcapsules (MCs) were used as a microscale engine to maneuver the fluid flow. The fluid actuation was tuned by various parameters such as substrate concentration, reaction rate, diameter of MCs and the population of the MCs inside the flow chamber. The same MCs, when suspended in a solution, showed buoyancy driven motility by creating oxygen bubbles via an enzymatic reaction and the velocity of the MCs was directly dependent on the number of nucleated oxygen bubbles generated on the MC surface.

Modulation of liquid structure and controlling molecular diffusion using supramolecular constructs.

The non-equilibrium liquid structure was achieved by interfacial jamming of pillar[5]arene carboxylic acid (P[5]AA) mediated by hydrogen bonding interactions. The assembly was reversibly modulated via jamming to unjamming transition thus dynamically shaping the liquid droplets. Interestingly, these supramolecular constructs showed pH-switchable gated diffusion of encapsulants, hence showcasing a next generation smart release system.

Joint Degeneration in a Mouse Model of Pseudoachondroplasia: ER Stress, Inflammation, and Block of Autophagy.

Pseudoachondroplasia (PSACH), a short limb skeletal dysplasia associated with premature joint degeneration, is caused by misfolding mutations in cartilage oligomeric matrix protein (COMP). Here, we define mutant-COMP-induced stress mechanisms that occur in articular chondrocytes of MT-COMP mice, a murine model of PSACH. The accumulation of mutant-COMP in the ER occurred early in MT-COMP articular chondrocytes and stimulated inflammation (TNFα) at 4 weeks, and articular chondrocyte death increased at 8 weeks while ER stress through CHOP was elevated by 12 weeks. Importantly, blockage of autophagy (pS6), the major mechanism that clears the ER, sustained cellular stress in MT-COMP articular chondrocytes. Degeneration of MT-COMP articular cartilage was similar to that observed in PSACH and was associated with increased MMPs, a family of degradative enzymes. Moreover, chronic cellular stresses stimulated senescence. Senescence-associated secretory phenotype (SASP) may play a role in generating and propagating a pro-degradative environment in the MT-COMP murine joint. The loss of CHOP or resveratrol treatment from birth preserved joint health in MT-COMP mice. Taken together, these results indicate that ER stress/CHOP signaling and autophagy blockage are central to mutant-COMP joint degeneration, and MT-COMP mice joint health can be preserved by decreasing articular chondrocyte stress. Future joint sparing therapeutics for PSACH may include resveratrol.

Valveless flow reversal by a pH responsive supramolecular micropump.

A valveless micropump was designed via dynamic supramolecular interaction between beta-cyclodextrin (ß-CD) and benzimidazole (BzI). It shows flow reversal in response to the pH change. An L-shaped microchannel was used to demonstrate the flow reversibility over long distances.

Maneuvering Fluid Motion and Flow-Induced Detection of Toxins by Enzyme Multilayer Films.

In view to develop an autonomous lab-on-a-chip device for detection of toxins without using any spectroscopic or electrochemical equipment, self-powered enzyme micropumps were fabricated via layer-by-layer assembly of enzymes and polyelectrolytes. The thin film-based enzyme micropumps turned on fluid flow in the presence of respective substrates in a concentration-dependent manner, and the rate of the enzymatic reaction was the key for maneuvering the fluid flow. Furthermore, the newly engineered enzyme-based micropumps were able to detect toxic metals and organophosphorus pesticides by modulating the fluid flow speed as the rate of the enzymatic reaction was altered by the presence of inhibitors. Thus, by regulating fluid flow in a micropump, low concentrations of analytes (e.g., target biomarkers and inhibitors) in biological fluids can be quantitatively identified for testing in a resource-constrained environment.

Pillar[5]arene microcapsules turn on fluid flow in the presence of paraquat.

We report the fabrication of pillar[5]arene (P[5]A) stabilized MCs via the self-assembly and crosslinking of P[5]A nanoaggregates at the liquid-liquid interface. These MC microengines turn on fluid flow in the presence of paraquat (PQ) due to "host-guest" molecular recognition. The microengines could be useful in designing non-mechanical micropumps, powerless microfluidics, and diagnostic devices.

Site-1 protease ablation in the osterix-lineage in mice results in bone marrow neutrophilia and hematopoietic stem cell alterations.

Site-1 protease (S1P) ablation in the osterix-lineage in mice drastically reduces bone development and downregulates bone marrow-derived skeletal stem cells. Here we show that these mice also suffer from spina bifida occulta with a characteristic lack of bone fusion in the posterior neural arches. Molecular analysis of bone marrow-derived non-red blood cell cells, via single-cell RNA-Seq and protein mass spectrometry, demonstrate that these mice have a much-altered bone marrow with a significant increase in neutrophils and Ly6C-expressing leukocytes. The molecular composition of bone marrow neutrophils is also different as they express more and additional members of the stefin A (Stfa) family of proteins. In vitro, recombinant Stfa1 and Stfa2 proteins have the ability to drastically inhibit osteogenic differentiation of bone marrow stromal cells, with no effect on adipogenic differentiation. FACS analysis of hematopoietic stem cells show that despite a decrease in hematopoietic stem cells, S1P ablation results in an increased production of granulocyte-macrophage progenitors, the precursors to neutrophils. These observations indicate that S1P has a role in the lineage specification of hematopoietic stem cells and/or their progenitors for development of a normal hematopoietic niche. Our study designates a fundamental requirement of S1P for maintaining a balanced regenerative capacity of the bone marrow niche.

One-Step Fabrication of Enzyme-Immobilized Reusable Polymerized Microcapsules from Microfluidic Droplets.

Enzyme immobilization is an essential prerequisite for biocatalysis. In this context, emulsion provides an excellent template for assembling enzymes at the oil-water interface. A microfluidic approach has been adopted to produce oil-in-water-type emulsions stabilized by gold nanoparticle-catalase conjugates. In situ ring-opening polymerization of the oil phase produces solid core enzyme-immobilized microcapsules (MCs). These resultant MCs exhibited a K m value of 42 mM and shows 1.1-fold higher activity compared to free enzymes. Finally, the robust MCs showed excellent recyclability, which can meet the demand of industrial biotechnological applications.