Bone-Targeting Exosome Mimetics Engineered by Bioorthogonal Surface Functionalization for Bone Tissue Engineering.

Extracellular vesicles have received a great interest as safe biocarriers in biomedical engineering. There is a need to develop more efficient delivery strategies to improve localized therapeutic efficacy and minimize off-target adverse effects. Here, exosome mimetics (EMs) are reported for bone targeting involving the introduction of hydroxyapatite-binding moieties through bioorthogonal functionalization. Bone-binding ability of the engineered EMs is verified with hydroxyapatite-coated scaffolds and an ex vivo bone-binding assay. The EM-bound construct provided a biocompatible substrate for cell adhesion, proliferation, and osteogenic differentiation. Particularly, the incorporation of Smoothened agonist (SAG) into EMs greatly increased the osteogenic capacity through the activation of hedgehog signaling. Furthermore, the scaffold integrated with EM/SAG significantly improved in vivo reossification. Lastly, biodistribution studies confirmed the accumulation of systemically administered EMs in bone tissue. This facile engineering strategy could be a versatile tool to promote bone regeneration, offering a promising nanomedicine approach to the sophisticated treatment of bone diseases.

Light-triggered thermal conductivity switching in azobenzene polymers.

Materials that can be switched between low and high thermal conductivity states would advance the control and conversion of thermal energy. Employing in situ time-domain thermoreflectance (TDTR) and in situ synchrotron X-ray scattering, we report a reversible, light-responsive azobenzene polymer that switches between high (0.35 W m-1 K-1) and low thermal conductivity (0.10 W m-1 K-1) states. This threefold change in the thermal conductivity is achieved by modulation of chain alignment resulted from the conformational transition between planar (trans) and nonplanar (cis) azobenzene groups under UV and green light illumination. This conformational transition leads to changes in the π-π stacking geometry and drives the crystal-to-liquid transition, which is fully reversible and occurs on a time scale of tens of seconds at room temperature. This result demonstrates an effective control of the thermophysical properties of polymers by modulating interchain π-π networks by light.

Nanostructured Lipid-based Films for Substrate Mediated Applications in Biotechnology.

Amphiphilic in nature, lipids spontaneously self-assemble into a range of nanostructures in the presence of water. Among lipid self-assembled structures, liposomes and supported lipid bilayers have long held scientific interest for their main applications in drug delivery and plasma membrane models, respectively. In contrast, lipid-based multi-layered membranes on solid supports only recently begun drawing scientists' attention. New studies on lipid films show that the stacking of multiple bilayers on a solid support yields interestingly complex features to these systems. Namely, multiple layers exhibit cooperative structural and dynamic behavior. In addition, the materials enable compartmentalization, templating, and enhanced release of several molecules of interest. Importantly, supported lipid phases exhibit long-range periodic nano-scale order and orientation that is tunable in response to a changing environment. Herein, we summarize current and pertinent understanding of lipid-based film research focusing on how unique structural characteristics enable the emergence of new applications in biotechnology including label-free biosensors, macroscale drug delivery, and substrate-mediated gene delivery. Our very recent contributions to lipid-based films, focusing on the structural characterization at the meso, nano, and molecular-scale, using Small-Angle X-ray Scattering, Atomic Force Microscopy, Photothermal Induced Resonance, and Solid-State NMR will be also highlighted.

Pyrolytic Carbon Nanosheets for Ultrafast and Ultrastable Sodium-Ion Storage.

Na-ion cointercalation in the graphite host structure in a glyme-based electrolyte represents a new possibility for using carbon-based materials (CMs) as anodes for Na-ion storage. However, local microstructures and nanoscale morphological features in CMs affect their electrochemical performances; they require intensive studies to achieve high levels of Na-ion storage performances. Here, pyrolytic carbon nanosheets (PCNs) composed of multitudinous graphitic nanocrystals are prepared from renewable bioresources by heating. In particular, PCN-2800 prepared by heating at 2800 °C has a distinctive sp2 carbon bonding nature, crystalline domain size of ≈44.2 Å, and high electrical conductivity of ≈320 S cm-1 , presenting significantly high rate capability at 600 C (60 A g-1 ) and stable cycling behaviors over 40 000 cycles as an anode for Na-ion storage. The results of this study show the unusual graphitization behaviors of a char-type carbon precursor and exceptionally high rate and cycling performances of the resulting graphitic material, PCN-2800, even surpassing those of supercapacitors.

Nanoscale partitioning of paclitaxel in hybrid lipid-polymer membranes.

Paclitaxel is a powerful drug against restenosis and many forms of cancer. However, its clinical application hinges on the ability to achieve suitable stabilized drug concentrations in an aqueous suspension while hindering drug crystallization. To engineer such formulations, it is imperative to understand paclitaxel's partitioning and crystallization within the carrier matrix. Lipid-polymer hybrid films have been recently shown to accommodate large paclitaxel loads and suppress crystallization. Additionally, such hybrid materials promote synergistic drug release compared to the pure constituents. Here, we leverage the composition sensitive photo-thermal induced resonance (PTIR) technique to study paclitaxel partitioning within hybrid films at the nanoscale. PTIR data reveal that paclitaxel nano-crystals segregate from lipid-only films but are well dispersed in polymer-only films. Remarkably, lipid-polymer hybrid films show enhanced partitioning of paclitaxel at the lipid-polymer phase boundaries, but still stifle crystallization, thus paving the way towards compositional and microstructural engineering of small-drug delivery systems.

Self-organization of Nucleic Acids in Lipid Constructs.

Lipids and nucleic acids (NAs) can hierarchically self-organize into a variety of nanostructures of increasingly complex geometries such as the 1D lamellar, 2D hexagonal, and 3D bicontinuous cubic phases. The diversity and complexity of those lipid-NA assemblies are interesting from a fundamental perspective as well as being relevant to the performance in gene delivery and gene silencing applications. The finding that not only the chemical make of the lipid-NA constructs, but their actual supramolecular organization, affects their gene transfection and silencing efficiencies has inspired physicists, chemists, and engineers to this field of research. At the moment it remains an open question how exactly the different lipid-NA structures interact with cells and organelles in order to output an optimal response. This article reviews our current understanding of the structures of different lipid-NA complexes and the corresponding cellular interaction mechanisms. The recent advances in designing optimal lipid-based NA carriers will be introduced with an emphasis on the structure-function relations.

Role of lipid polymorphism in acoustically sensitive liposomes.

Ultrasound (US) triggered drug release is a promising drug delivery method that allows ex vivo modulation of treatment intensity and duration. This method relies on the synergistic interaction between the rupture of sonosensitive particles and enhanced plasma membrane permeability. Conventional liposomal systems where the drug passively diffuses through the membrane show virtually no response to acoustic energy. One method to activate drug transport is to induce a topological restructuring of the lipid membrane (zero intrinsic curvature, H = 0) by puncturing pores (H < 0) through which the drug can readily leak out from the interior of the liposomes. In this work we demonstrate strategies to lower the energy cost of creating such membrane defects by introducing lipid molecules with molecular shapes prone to self-assemble into non-lamellar (negative intrinsic curvature, H < 0) structures. All formulations investigated comprise the relevant components typically required for delivery applications such as stealth moieties, cholesterol, and phospholipids. Small angle X-ray scattering studies of a number of lipid systems at increasing amounts of phosphatidylethanolamine (PE) phospholipids reveal that membranes without PE respond to ultrasound by thinning ca. 10 Å, which concomitantly lowers the bending rigidity quadratically in addition to increasing the passive drug permeability. However, at the appropriate PE content the lipid systems display a classic lamellar structure (H = 0) that undergoes a topological transformation after ultrasound exposure into lipid tubes of the reversed type (H < 0) packed in a 2D hexagonal array. At the dilute regime, Fluorescence Microscopy of giant unilamellar vesicles (GUVs) comprising DOPE also experience ultrasound induced restructuring that can be modulated by DOPE content. In general, smaller vesicles of diverse shape connect and form into a "pearl-necklace" configuration. We argue that the inclusion of DOPE within the GUV membrane may result in curvature-driven lipid sorting, providing the system with local membrane instabilities that drive vesicle pearling when exposed to ultrasound.

Self-Assembled Nanocomposite Hydrogels as Carriers for Demineralized Bone Matrix Particles and Enhanced Bone Repair.

Demineralized bone matrix (DBM) has been widely used as an allogeneic alternative to autologous bone graft for bone repair. However, more extensive use of DBM is limited due to its particulate nature after demineralization and rapid particle dispersion following irrigation, resulting in unpredictable osteoinductivity. Here, a new design of injectable hydrogel carriers for DBM that combine self-healing ability and osteogenic properties based on the self-assembly of guanidinylated hyaluronic acid and silica-rich nanoclays is reported. The nanoclays serve as reversible linkages to form a dynamic hydrogel network with the guanidine moieties on the polymer chains. Gelation kinetics and mechanical properties can be controlled by altering nanoclay content in the hydrogel. The resulting hydrogel exerts self-healing ability due to its dynamic crosslinks and well retains its overall performance with high DBM loading. The hydrogel exhibits great cytocompatibility and osteogenic effects mediated by the nanoclays. In vivo delivery of DBM using the nanocomposite hydrogel further demonstrates robust bone regeneration in a mouse calvarial defect model in comparison to DBM delivered with aqueous HA. This work suggests a promising hydrogel platform for many applications including therapeutic delivery and tissue engineering.

Black Phosphorus-Based Dynamic Self-Healing Hydrogel to Integrate Demineralized Bone Matrix and Noggin-Targeting siRNA for Synergistic Osteogenesis.

Although a demineralized bone matrix (DBM) is often used as an alternative to an autologous bone graft, its clinical application is still hampered by easy dispersion of DBM particles and insufficient osteoinductivity in the defect site. Herein, we designed a self-healing hydrogel for DBM that can rapidly restore its structural integrity after damage based on amino-rich black phosphorus (BP) nanosheets and aldehyde-functionalized hyaluronic acid (AHA). Given the increased expression of bone morphogenetic protein (BMP) antagonists by DBM stimulation, the osteogenic potency of DBM in the hydrogel carrier was further enhanced by abrogating the BMP antagonism. The BP/AHA hydrogel provided dynamic polymer-nanosheet networks that combine injectability, modability, and physical stability with high DBM loading, where the BP nanosheets served as osteogenic cross-linkers to promote biomineralization and deliver siRNA to suppress undesirable expression of BMP antagonist noggin by DBM. As a result, the BP/AHA hydrogel integrated with DBM and noggin-targeting siRNA synergistically promoted osteogenic differentiation of mesenchymal stem cells by enhancing BMP/Smad signaling. This work demonstrates a promising strategy to improve the efficacy of bone regeneration using bone graft.

Complementary modulation of BMP signaling improves bone healing efficiency.

The bone morphogenetic protein (BMP) signaling pathway plays a crucial role in bone development and regeneration. While BMP-2 is widely used as an alternative to autograft, its clinical application has raised concerns about adverse side effects and deteriorated bone quality. Therefore, there is a need to develop more sophisticated approaches to regulate BMP signaling and promote bone regeneration. Here, we present a novel complementary strategy that targets both BMP antagonist noggin and agonist Trb3 to enhance bone defect repair without the application of exogenous BMP-2. In vitro studies showed that overexpression of Trb3 with simultaneous noggin suppression significantly promotes osteogenic differentiation of mesenchymal stem cells. This was accompanied by increased BMP/Smad signaling. We also developed sterosome nanocarriers, a non-phospholipid liposomal system, to achieve non-viral mediated noggin suppression and Trb3 overexpression. The gene-loaded sterosomes were integrated onto an apatite-coated polymer scaffold for in vivo calvarial defect implantation, resulting in robust bone healing compared to BMP-2 treatments. Our work provides a promising alternative for high-quality bone formation by regulating expression of BMP agonists and antagonists.

Lifting gate polydimethylsiloxane microvalves and pumps for microfluidic control.

We describe the development and characterization of pneumatically actuated "lifting gate" microvalves and pumps. A fluidic layer containing the gate structure and a pneumatic layer are fabricated by soft-lithography in PDMS and bonded permanently with an oxygen plasma treatment. The microvalve structures are then reversibly bonded to a featureless glass or plastic substrate to form hybrid glass-PDMS and plastic-PDMS microchannel structures. The break-through pressures of the microvalve increase linearly up to 65 kPa as the closing pressure increases. The pumping capability of these structures ranges from the nanoliter to microliter scale depending on the number of cycles and closing pressure employed. The micropump structures exhibit up to 86.2% pumping efficiency from flow rate measurements. The utility of these structures for integrated sample processing is demonstrated by performing an automated immunoassay. These lifting gate valve and pump structures enable facile integration of complex microfluidic control systems with a wide range of lab-on-a-chip substrates.

Role of tropical lower stratosphere winds in quasi-biennial oscillation disruptions.

In 2016, the westerly quasi-biennial oscillation (WQBO) in the equatorial stratosphere was unprecedentedly disrupted by westward forcing near 40 hPa; this was followed by another disruption in 2020. Strong extratropical Rossby waves propagating toward the tropics were considered the main cause of the disruptions, but why the zonal wind is reversed only in the middle of the WQBO remains unclear. Here, we show that strong westerly winds in the equatorial lower stratosphere (70 to 100 hPa) help to disrupt the WQBO by hindering the wind reversal at its base. They also help equatorial westward waves propagate further upward, increasing the negative forcing at around 40 hPa that drives the QBO disruptions. Tropical westerly winds have been increasing in the past and are projected to increase in a warmer climate. These background wind changes may allow more frequent QBO disruptions in the future, leading to less predictability in atmospheric weather and climate systems.

Bioactive Scaffolds Integrated with Liposomal or Extracellular Vesicles for Bone Regeneration.

With population aging and increased life expectancy, an increasing number of people are facing musculoskeletal health problems that necessitate therapeutic intervention at defect sites. Bone tissue engineering (BTE) has become a promising approach for bone graft substitutes as traditional treatments using autografts or allografts involve clinical complications. Significant advancements have been made in developing ideal BTE scaffolds that can integrate bioactive molecules promoting robust bone repair. Herein, we review bioactive scaffolds tuned for local bone regenerative therapy, particularly through integrating synthetic liposomal vesicles or extracellular vesicles to the scaffolds. Liposomes offer an excellent drug delivery system providing sustained release of the loaded bioactive molecules. Extracellular vesicles, with their inherent capacity to carry bioactive molecules, are emerging as an advanced substitute of synthetic nanoparticles and a novel cell-free therapy for bone regeneration. We discuss the recent advance in the use of synthetic liposomes and extracellular vesicles as bioactive materials combined with scaffolds, highlighting major challenges and opportunities for their applications in bone regeneration. We put a particular focus on strategies to integrate vesicles to various biomaterial scaffolds and introduce the latest advances in achieving sustained release of bioactive molecules from the vesicle-loaded scaffolds at the bone defect site.

Three-Dimensional Microphase Separation and Synergistic Permeability in Stacked Lipid-Polymer Hybrid Membranes.

We present new structures of soft-material thin films that augment the functionality of substrate-mediated delivery systems. A hybrid material composed of phospholipids and block copolymers adopts a multilayered membrane structure supported on a solid surface. The hybrid films comprise intentional intramembrane heterogeneities that register across multilayers. These stacked domains convey unprecedented enhancement and control of permeability of solutes across micrometer-thick films. Using grazing incidence X-ray scattering, phase contrast atomic force microscopy, and confocal microscopy, we observed that in each lamella, lipid and polymers partition unevenly within the membrane plane segregating into lipid- or polymer-rich domains. Interestingly, we found evidence that like-domains align in registry across multilayers, thereby making phase separation three-dimensional. Phase boundaries exist over extended length scales to compensate the height mismatch between lipid and polymer molecules. We show that microphase separation in hybrid films can be exploited to augment the capability of drug-eluting substrates. Lipid-polymer hybrid films loaded with paclitaxel show synergistic permeability of drug compared to single-component counterparts. We present a thorough structural study of stacked lipid-polymer hybrid membranes and propose that the presence of registered domains and domain boundaries impart enhanced drug release functionality. This work offers new perspectives in designing thin films for controlled delivery applications.

Thermally Functional Liquid Crystal Networks by Magnetic Field Driven Molecular Orientation.

Aligned liquid crystal networks were synthesized by photopolymerization of liquid crystal monomers in the presence of magnetic fields. Grazing incident wide-angle X-ray scattering was used to characterize the degree of molecular alignment of mesogen chains and time-domain thermoreflectance was used to measure thermal conductivity. Liquid crystal networks with mesogenic units aligned perpendicular and parallel to the substrate exhibit thermal conductivity of 0.34 W m-1 K-1 and 0.22 W m-1 K-1, respectively. The thermal conductivity and orientational order of liquid crystal networks vary as a function of temperature. The thermal conductivity of liquid crystal networks can be manipulated by a magnetic field at above the glass transition temperature (65 °C) where the reduced viscosity enables molecular reorientation on the time scale of 10 min.

Citrus-Peel-Derived, Nanoporous Carbon Nanosheets Containing Redox-Active Heteroatoms for Sodium-Ion Storage.

Advanced design of nanostructured functional carbon materials for use in sustainable energy storage systems suffers from complex fabrication procedures and the use of special methods and/or expensive precursors, limiting their practical applications. In this study, nanoporous carbon nanosheets (NP-CNSs) containing numerous redox-active heteroatoms (C/O and C/N ratios of 5.5 and 34.3, respectively) were fabricated from citrus peels by simply heating the peels in the presence of potassium ions. The NP-CNSs had a 2D-like morphology with a high aspect ratio of >100, high specific surface area of 1167 m(2) g(-1), and a large amount of nanopores between 1 and 5 nm. The NP-CNSs also had an electrical conductivity of 2.6 × 10(1) s cm(-1), which is approximately 50 times higher than that of reduced graphene oxide. These unique material properties resulted in superior electrochemical performance with a high specific capacity of 140 mAh g(-1) in the cathodic potential range. In addition, symmetric full-cell devices based on the NP-CNSs showed excellent cyclic performance over 100,000 repetitive cycles.