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.

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.

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.

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.

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.

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.

Room-Temperature Synthesis of Germanium Oxide Nanofilaments and Their Potential Use as Luminescent Self-Cleaning Surfaces.

Germanium oxide nanofilaments (GNFs) have been synthesized under ambient conditions from the gas phase using germanium tetrachloride as a precursor. Non-crystalline GNFs synthesized by this procedure are 1-10 µm in length and 80-110 nm in diameter applying Droplet Assisted Growth and Shaping (DAGS) Chemistry. The relative humidity has been adjusted at various values in order to demonstrate the crucial role of humidity in the gas phase for the nanofilament synthesis. The novel GNFs show a strong luminescence emission in the ultra-violet and light blue region. In addition, a self-cleaning and superhydrophobic properties could be introduced in the luminescent GNF nanofilaments by simple treatment with silane molecules.

Directed In Situ Shaping of Complex Nano- and Microstructures during Chemical Synthesis.

Chemical composition and shape determine the basic properties of any object. Commonly, chemical synthesis and shaping follow each other in a sequence, although their combination into a single process would be an elegant simplification. Here, a pathway of simultaneous synthesis and shaping as applied to polysiloxanes on the micro- and nanoscale is presented. Complex structures such as stars, chalices, helices, volcanoes, rods, or combinations thereof are obtained. Varying the shape-controlling reaction parameters including temperature, water saturation, and the type of substrate allows to direct the reaction toward specific structures. A general mechanism of growth is suggested and analytical evidence and thermodynamic calculations to support it are provided. An aqueous droplet in either gaseous atmosphere or in a liquid organic solvent serves as a spatially confined polymerization volume. By substituting the starting materials, germanium-based nanostructures are also obtained. This transferability marks this approach as a major step toward a generally applicable method of chemical synthesis including in situ shaping.

Transcriptome analysis of injured human meniscus reveals a distinct phenotype of meniscus degeneration with aging.

OBJECTIVE: Meniscus tears are associated with a heightened risk of osteoarthritis. This study aimed to advance our understanding of the metabolic state of injured human meniscus at the time of arthroscopic partial meniscectomy through transcriptome-wide analysis of gene expression in relation to the patient's age and degree of cartilage chondrosis. METHODS: The degree of chondrosis of knee cartilage was recorded at the time of meniscectomy in symptomatic patients without radiographic osteoarthritis. RNA preparations from resected menisci (n = 12) were subjected to transcriptome-wide microarray and QuantiGene Plex analyses. Variations in the relative changes in gene expression with age and chondrosis were analyzed, and integrated biologic processes were investigated computationally. RESULTS: We identified a set of genes in torn menisci that were differentially expressed with age and chondrosis. There were 866 genes that were differentially regulated (≥1.5-fold difference and P < 0.05) with age and 49 with chondrosis. In older patients, genes associated with cartilage and skeletal development and extracellular matrix synthesis were repressed, while those involved in immune response, inflammation, cell cycle, and cellular proliferation were stimulated. With chondrosis, genes representing cell catabolism (cAMP catabolic process) and tissue and endothelial cell development were repressed, and those involved in T cell differentiation and apoptosis were elevated. CONCLUSION: Differences in age-related gene expression suggest that in older adults, meniscal cells might dedifferentiate and initiate a proliferative phenotype. Conversely, meniscal cells in younger patients appear to respond to injury, but they maintain the differentiated phenotype. Definitive molecular signatures identified in damaged meniscus could be segregated largely with age and, to a lesser extent, with chondrosis.

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.

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.

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.

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.

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.

Site-1 protease is essential to growth plate maintenance and is a critical regulator of chondrocyte hypertrophic differentiation in postnatal mice.

Site-1 protease (S1P) is a proprotein convertase with essential functions in lipid homeostasis and unfolded protein response pathways. We previously studied a mouse model of cartilage-specific knock-out of S1P in chondroprogenitor cells. These mice exhibited a defective cartilage matrix devoid of type II collagen protein (Col II) and displayed chondrodysplasia with no endochondral bone formation even though the molecular program for endochondral bone development appeared intact. To gain insights into S1P function, we generated and studied a mouse model in which S1P is ablated in postnatal chondrocytes. Postnatal ablation of S1P results in chondrodysplasia. However, unlike early embryonic ablations, the growth plates of these mice exhibit a lack of Ihh, PTHrP-R, and Col10 expression indicating a loss of chondrocyte hypertrophic differentiation and thus disruption of the molecular program required for endochondral bone development. S1P ablation results in rapid growth plate disruption due to intracellular Col II entrapment concomitant with loss of chondrocyte hypertrophy suggesting that these two processes are related. Entrapment of Col II in the chondrocytes of the prospective secondary ossification center precludes its development. Trabecular bone formation is dramatically diminished in the primary spongiosa and is eventually lost. The primary growth plate is eradicated by apoptosis but is gradually replaced by a fully functional new growth plate from progenitor stem cells capable of supporting new bone growth. Our study thus demonstrates that S1P has fundamental roles in the preservation of postnatal growth plate through chondrocyte differentiation and Col II deposition and functions to couple growth plate maturation to trabecular bone development in growing mice.

Antiangiogenic and anticancer molecules in cartilage.

Cartilage is one of the very few naturally occurring avascular tissues where lack of angiogenesis is the guiding principle for its structure and function. This has attracted investigators who have sought to understand the biochemical basis for its avascular nature, hypothesising that it could be used in designing therapies for treating cancer and related malignancies in humans through antiangiogenic applications. Cartilage encompasses primarily a specialised extracellular matrix synthesised by chondrocytes that is both complex and unique as a result of the myriad molecules of which it is composed. Of these components, a few such as thrombospondin-1, chondromodulin-1, the type XVIII-derived endostatin, SPARC (secreted protein acidic and rich in cysteine) and the type II collagen-derived N-terminal propeptide (PIIBNP) have demonstrated antiangiogenic or antitumour properties in vitro and in vivo preclinical trials that involve several complicated mechanisms that are not completely understood. Thrombospondin-1, endostatin and the shark-cartilage-derived Neovastat preparation have also been investigated in human clinical trials to treat several different kinds of cancers, where, despite the tremendous success seen in preclinical trials, these molecules are yet to show success as anticancer agents. This review summarises the current state-of-the-art antiangiogenic characterisation of these molecules, highlights their most promising aspects and evaluates the future of these molecules in antiangiogenic applications.

Control of surface tension at liquid-liquid interfaces using nanoparticles and nanoparticle-protein complexes.

Subtle changes in the monolayer structure of nanoparticles (NPs) influence the interfacial behavior of both NPs and NP-protein conjugates. In this study, we use a series of monolayer-protected gold NPs to explore the role of particle hydrophobicity on their dynamic behavior at the toluene-water interface. Using dynamic surface tension measurements, we observed a linear decrease in the meso-equilibrium surface tension (γ) and faster dynamics as the hydrophobicity of the ligands increases. Further modulation of γ is observed for the corresponding NP-protein complexes at the charge-neutralization point.

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.

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.

Recent advances in biomarkers in osteoarthritis.

PURPOSE OF REVIEW: Osteoarthritis is a joint disease characterized by a nonsymptomatic, preradiographical phase that if distinguished would allow earlier osteoarthritis diagnosis. Biochemical biomarkers offer a potential nonradiographical alternative to detect early, nonsymptomatic osteoarthritis. RECENT FINDINGS: Biomarker development for osteoarthritis diagnosis is still in the forefront of the research repertoire in osteoarthritis. A number of previously identified biomarkers derived from cartilage breakdown or enzymes that cause cartilage degeneration still have prominence and are now better characterized with increasing use in identifying disease severity, progression, and testing treatment options. Combinations of cartilage-derived and bone-derived biomarkers have been used to subgroup osteoarthritis patients that could impact treatment and address the importance of bone turnover in cartilage integrity. Increasingly, inflammation markers have been used to profile osteoarthritis progression attesting to the inflammatory nature of osteoarthritis. The application of proteomic technologies has generated several new, nonconventional biomarkers that could allow better profiling of osteoarthritis. SUMMARY: Biomarker combinations have the ability to subgroup the heterogenous osteoarthritis population to allow a better scrutiny of diagnosis and treatment options. The application of different technological platforms to osteoarthritis would allow a better understanding of its pathology and could provide for appropriate candidates for earlier detection of osteoarthritis.