Induced Endothelial Cell-Integrated Liver Assembloids Promote Hepatic Maturation and Therapeutic Effect on Cholestatic Liver Fibrosis.

The transplantation of pluripotent stem cell (PSC)-derived liver organoids has been studied to solve the current donor shortage. However, the differentiation of unintended cell populations, difficulty in generating multi-lineage organoids, and tumorigenicity of PSC-derived organoids are challenges. However, direct conversion technology has allowed for the generation lineage-restricted induced stem cells from somatic cells bypassing the pluripotent state, thereby eliminating tumorigenic risks. Here, liver assembloids (iHEAs) were generated by integrating induced endothelial cells (iECs) into the liver organoids (iHLOs) generated with induced hepatic stem cells (iHepSCs). Liver assembloids showed enhanced functional maturity compared to iHLOs in vitro and improved therapeutic effects on cholestatic liver fibrosis animals in vivo. Mechanistically, FN1 expressed from iECs led to the upregulation of Itgα5/ß1 and Hnf4α in iHEAs and were correlated to the decreased expression of genes related to hepatic stellate cell activation such as Lox and Spp1 in the cholestatic liver fibrosis animals. In conclusion, our study demonstrates the possibility of generating transplantable iHEAs with directly converted cells, and our results evidence that integrating iECs allows iHEAs to have enhanced hepatic maturation compared to iHLOs.

Effect of Waste Ceramic Powder on the Properties of Alkali-Activated Slag and Fly Ash Pastes Exposed to High Temperature.

This paper presents the effects of alkali-activated blast furnace slag and fly ash (AASF) paste added with waste ceramic powder (WCP) on mechanical properties, weight loss, mesoscopic cracks, reaction products, and microstructure when exposed to 300, 600, and 900 °C. Using waste ceramic powder to replace blast furnace slag and fly ash, the replacement rate was 0-20%. The samples cured at 45 °C for 28 days were heated to 300, 600, and 900 °C to determine the residual compressive strength and weight loss at the relevant temperature. We evaluated the deterioration of the paste at each temperature through mesoscopic images, ultrasonic pulse velocity (UPV), thermogravimetric analysis (TG), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and with a scanning electron microscope (SEM). Relevant experimental results show that: (1) with the increase in waste ceramic powder content, the compressive strength of samples at various temperatures increased, and at 300 °C, the compressive strength of all the samples reached the highest value; (2) the residual weight increased with the increase in the content of the waste ceramic powder; (3) with a further increase in temperature, all the samples produced more mesoscopic cracks; (4) at each temperature, with the rise in waste ceramic powder content, the value of the ultrasonic pulse velocity increased; (5) the TG results showed that, as the content of waste ceramic powder increased, the formation of C-A-S-H gel and hydrotalcite decreased; (6) XRD and FTIR spectra showed that, at 900 °C, the use of waste ceramic powder reduced the formation of harmful crystalline phases; (7) the SEM image showed that, at 900 °C, as the content of waste ceramic powder increased, the compactness of the sample was improved. In summary, the addition of waste ceramic powder can improve the mechanical properties of the alkali-activated paste at high temperatures, reduce the occurrence of cracks, and make the microstructure denser.

Implementation of Combinatorial Genetic and Microenvironmental Engineering to Microbial-Based Field-Deployable Microbead Biosensors for Highly Sensitive and Remote Chemical Detection.

Bioreporters, microbial species genetically engineered to provide measurable signals in response to specific chemicals, have been widely investigated as sensors for biomedical and environmental monitoring. More specifically, the bioreporter encapsulated within a biocompatible material, such as a hydrogel that can provide a suitable microenvironment for its prolonged activity as well as efficient scalable production, has been viewed as a more broadly applicable mode of biosensors. In this study, alginate-based microbeads encapsulated with the bacterial bioreporter capable of expressing green fluorescence protein in response to nitro compounds (e.g., trinitrotoluene and dinitrotoluene) are developed as biosensors. To significantly enhance the sensitivity of the microbial-based microbead biosensors, "multifaceted" modification strategies are simultaneously employed: (1) multiple genetic modifications of the bioreporter, (2) tuning the physicomechanical properties of the encapsulating microbeads, (3) controlling the initial cell density within the microbeads, and (4) enrichment of nitro compounds inside microbeads via functional nanomaterials. These microbial and microenvironmental engineering approaches combine to significantly enhance the sensing capability, even allowing highly sensitive remote detection under a low-vapor phase. Thus, the strategy developed herein is expected to contribute to various cell-based biosensors.

Measurement of multispectral scattering properties in mouse brain tissue.

We present the scattering properties of mouse brain using multispectral diffraction phase microscopy. Typical diffraction phase microscopy was incorporated with the broadband light source which offers the measurement of the scattering coefficient and anisotropy in the spectral range of 550-900 nm. The regional analysis was performed for both the myeloarchitecture and cytoarchitecture of the brain tissue. Our results clearly evaluate the multispectral scattering properties in the olfactory bulb and corpus callosum. The scattering coefficient measured in the corpus callosum is about four times higher than in the olfactory bulb. It also indicates that it is feasible to realize the quantitative phase microscope in near infrared region for thick brain tissue imaging.

Mitochondria localization induced self-assembly of peptide amphiphiles for cellular dysfunction.

Achieving spatiotemporal control of molecular self-assembly associated with actuation of biological functions inside living cells remains a challenge owing to the complexity of the cellular environments and the lack of characterization tools. We present, for the first time, the organelle-localized self-assembly of a peptide amphiphile as a powerful strategy for controlling cellular fate. A phenylalanine dipeptide (FF) with a mitochondria-targeting moiety, triphenyl phosphonium (Mito-FF), preferentially accumulates inside mitochondria and reaches the critical aggregation concentration to form a fibrous nanostructure, which is monitored by confocal laser scanning microscopy and transmission electron microscopy. The Mito-FF fibrils induce mitochondrial dysfunction via membrane disruption to cause apoptosis. The organelle-specific supramolecular system provides a new opportunity for therapeutics and in-depth investigations of cellular functions.Spatiotemporal control of intracellular molecular self-assembly holds promise for therapeutic applications. Here the authors develop a peptide consisting of a phenylalanine dipeptide with a mitochondrial targeting moiety to form self-assembling fibrous nanostructures within mitochondria, leading to apoptosis.

A two-photon fluorescent probe for lysosomal zinc ions.

The selective detection of zinc ions in lysosomes over that in cytosol is achieved with a fluorescent probe, which enabled the fluorescence imaging of endogenous zinc ions in lysosomes of NIH 3T3 cells as well as mouse hippocampal tissues by two-photon microscopy under excitation at 900 nm.

Ligand Accessibility and Bioactivity of a Hormone-Dendrimer Conjugate Depend on pH and pH History.

Estrogen conjugates with a polyamidoamine (PAMAM) dendrimer have shown remarkably selective regulation of the nongenomic actions of estrogens in target cells. In response to pH changes, however, these estrogen-dendrimer conjugates (EDCs) display a major morphological transition that alters the accessibility of the estrogen ligands that compromises the bioactivity of the EDC. A sharp break in dynamic behavior near pH 7 occurs for three different ligands on the surface of a PAMAM-G6 dendrimer: a fluorophore (tetramethylrhodamine [TMR]) and two estrogens (17α-ethynylestradiol and diphenolic acid). Collisional quenching and time-resolved fluorescence anisotropy experiments with TMR-PAMAM revealed high ligand shielding above pH 7 and low shielding below pH 7. Furthermore, when the pH was cycled from 8.5 (conditions of ligand-PAMAM conjugation) to 4.5 (e.g., endosome/lysosome) and through 6.5 (e.g., hypoxic environment) back to pH 8.5, the 17α-ethynylestradiol- and diphenolic acid-PAMAM conjugates experienced a dramatic, irreversible loss in cell stimulatory activity; dynamic NMR studies indicated that the hormonal ligands had become occluded within the more hydrophobic core of the PAMAM dendrimer. Thus, the active state of these estrogen-dendrimer conjugates appears to be metastable. This pH-dependent irreversible masking of activity is of considerable relevance to the design of drug conjugates with amine-bearing PAMAM dendrimers.

Colloidal superstructures programmed into magnetic Janus particles.

By engineering thin magnetic films onto homogeneous colloidal particles, various crystalline lattices are induced from simple magnetic Janus spheres. In situ formation of dicolloids amplifies the diversity of achievable dynamic structures. The competition between shape anisotropy and dipole orientation generates mesoscopic isomerism. This opens design space for anisotropic building blocks for smart colloidal materials.

Single-molecule observation of long jumps in polymer adsorption.

Single-molecule fluorescence imaging of adsorption onto initially bare surfaces shows that polymer chains need not localize immediately after arrival. In a system optimized to present limited adsorption sites (quartz surface to which polyethylene glycol (PEG) chains adsorb from aqueous solution at pH 8.2), we find that some chains diffuse back into bulk solution and readsorb at some distance away, sometimes multiple times before they either localize at a stable position or diffuse away into bulk solution. This mechanism of surface diffusion is considerably more rapid than the classical model in which adsorbed polymers crawl on surfaces while the entire molecule remains adsorbed, suggesting the conceptual generality of a recent report ( Phys. Rev. Lett. 2013 , 110 , 256101 ) but in a new experimental system and with comparison of different chain lengths. We find the trajectories with jumps to follow a truncated Lévy distribution of step size with limiting slope -2.5, consistent with a well-defined, rapid surface diffusion coefficient over the times we observe. The broad waiting time distribution appears to reflect that polymer chains possess a broad distribution of bound fraction: the smaller the bound fraction of a given chain, the shorter the surface residence time before executing the next surface jump.

Modular stitching to image single-molecule DNA transport.

For study of time-dependent conformation, all previous single-molecule imaging studies of polymer transport involve fluorescence labeling uniformly along the chain, which suffers from limited resolution due to the diffraction limit. Here we demonstrate the concept of submolecular single-molecule imaging with DNA chains assembled from DNA fragments such that a chain is labeled at designated spots with covalently attached fluorescent dyes and the chain backbone with dyes of different color. High density of dyes ensures good signal-to-noise ratio to localize the designated spots in real time with nanometer precision and prevents significant photobleaching for long-time tracking purposes. To demonstrate usefulness of this approach, we image electrophoretic transport of λ-DNA through agarose gels. The unexpected pattern is observed that one end of each molecule tends to stretch out in the electric field while the other end remains quiescent for some time before it snaps forward and the stretch-recoil cycle repeats. These features are neither predicted by prevailing theories of electrophoresis mechanism nor detectable by conventional whole-chain labeling methods, which demonstrate pragmatically the usefulness of modular stitching to reveal internal chain dynamics of single molecules.

Linking synchronization to self-assembly using magnetic Janus colloids.

Synchronization occurs widely in the natural and technological worlds, from the rhythm of applause and neuron firing to the quantum mechanics of coupled Josephson junctions, but has not been used to produce new spatial structures. Our understanding of self-assembly has evolved independently in the fields of chemistry and materials, and with a few notable exceptions has focused on equilibrium rather than dynamical systems. Here we combine these two phenomena to create synchronization-selected microtubes of Janus colloids, micron-sized spherical particles with different surface chemistry on their opposing hemispheres, which we study using imaging and computer simulation. A thin nickel film coats one hemisphere of each silica particle to generate a discoid magnetic symmetry, such that in a precessing magnetic field its dynamics retain crucial phase freedom. Synchronizing their motion, these Janus spheres self-organize into micrometre-scale tubes in which the constituent particles rotate and oscillate continuously. In addition, the microtube must be tidally locked to the particles, that is, the particles must maintain their orientation within the rotating microtube. This requirement leads to a synchronization-induced structural transition that offers various applications based on the potential to form, disintegrate and fine-tune self-assembled in-motion structures in situ. Furthermore, it offers a generalizable method of controlling structure using dynamic synchronization criteria rather than static energy minimization, and of designing new field-driven microscale devices in which components do not slavishly follow the external field.

Janus and multiblock colloidal particles.

We review recent developments in the synthesis and self-assembly of Janus and multiblock colloidal particles, highlighting new opportunities for colloid science and technology that are enabled by encoding orientational order between particles as they self-assemble. Emphasizing the concepts of molecular colloids and colloid valence unique to such colloids, we describe their rational self-assembly into colloidal clusters, taking monodisperse tetrahedra as an example. We also introduce a simple method to lock clusters into permanent shapes. Extending this to 2D lattices, we also review recent progress in assembling new open colloidal networks including the kagome lattice. In each application, areas of opportunity are emphasized.

Staged self-assembly of colloidal metastructures.

We demonstrate sequential assembly of chemically patchy colloids such that their valence differs from stage to stage to produce hierarchical structures. For proof of concept, we employ ACB triblock spheres suspended in water, with the C middle band electrostatically repulsive. In the first assembly stage, only A-A hydrophobic attraction contributes, and discrete clusters form. They can be stored, but subsequently activated to allow B-B attractions, leading to higher-order assembly of clusters with one another. The growth dynamics, observed at a single particle level by fluorescence optical microscopy, obey the kinetics of stepwise polymerization, forming chains, pores, and networks. Between linked clusters, we identify three possible bond geometries, linear, triangular, and square, by an argument that is generalizable to other patchy colloid systems. This staged assembly strategy offers a promising route to fabricate colloidal assemblies bearing multiple levels of structural and functional complexity.

Colloidal rotation near the colloidal glass transition.

We compare, using single-particle optical imaging, trajectories of rotation and translation for micron-sized spheres in index-matched colloidal suspensions near their glass transition. Rotational trajectories, while they show intermittent caged behavior associated with supercooled and glassy behavior, explore a sufficiently wider phase space such that in the averaged mean-square angular displacement there appears no plateau regime, but instead sub-Fickian angular diffusion that follows an apparent power law in time. We infer translation and rotation time constants, the former being the time to diffuse a particle diameter and the latter being the time to rotate a full revolution. Correlation between time constants increases with increasing volume fraction, but unlike the case for molecular glasses, the rotation time constant slows more weakly than the translation time.

Triblock colloids for directed self-assembly.

Methods for functionalizing micrometer-sized colloidal spheres with three or more zones of chemical functionality (ABA or ABC) are described. To produce ABA triblock colloids, we functionalized the north pole, south pole, and equator to produce what we call X, Y, and K functionality according to the number of allowed nearest neighbors and their spatial arrangements. These synthesis methods allowed targeting of various lattice structures whose bonding between neighboring particles in liquid suspension was visualized in situ by optical microscopy.

How liposomes diffuse in concentrated liposome suspensions.

Building upon the observation that liposomes of zwitterionic lipids can be stabilized against fusion by the adsorption of cationic nanoparticles (Yu, Y.; Anthony, S.; Zhang, L.; Bae, S. C.; Granick, S. J. Phys. Chem. C2007, 111, 8233), we study, using single-particle fluorescence tracking, mobility in this distinctively deformable colloid system, in the volume fraction range of φ = 0.01 to 0.7. Liposome motion is diffusive and homogeneous at low volume fractions, but separable fast and slow populations emerge as the volume fraction increases beyond φ ≈ 0.45, the same volume fraction at which hard colloids with sufficiently strong attraction are known to experience gelation. This is reflected not only in scaling of the mean square displacement, but also in the step size distribution (van Hove function) measured by fluorescence imaging. The fast liposomes are observed to follow Brownian motion, and the slow ones follow anomalous diffusion characterized by a 1/3 time scaling of their mean square displacement.

Directed self-assembly of a colloidal kagome lattice.

A challenging goal in materials chemistry and physics is spontaneously to form intended superstructures from designed building blocks. In fields such as crystal engineering and the design of porous materials, this typically involves building blocks of organic molecules, sometimes operating together with metallic ions or clusters. The translation of such ideas to nanoparticles and colloidal-sized building blocks would potentially open doors to new materials and new properties, but the pathways to achieve this goal are still undetermined. Here we show how colloidal spheres can be induced to self-assemble into a complex predetermined colloidal crystal-in this case a colloidal kagome lattice-through decoration of their surfaces with a simple pattern of hydrophobic domains. The building blocks are simple micrometre-sized spheres with interactions (electrostatic repulsion in the middle, hydrophobic attraction at the poles, which we call 'triblock Janus') that are also simple, but the self-assembly of the spheres into an open kagome structure contrasts with previously known close-packed periodic arrangements of spheres. This open network is of interest for several theoretical reasons. With a view to possible enhanced functionality, the resulting lattice structure possesses two families of pores, one that is hydrophobic on the rims of the pores and another that is hydrophilic. This strategy of 'convergent' self-assembly from easily fabricated colloidal building blocks encodes the target supracolloidal architecture, not in localized attractive spots but instead in large redundantly attractive regions, and can be extended to form other supracolloidal networks.

Supracolloidal reaction kinetics of Janus spheres.

Clusters in the form of aggregates of a small number of elemental units display structural, thermodynamic, and dynamic properties different from those of bulk materials. We studied the kinetic pathways of self-assembly of "Janus spheres" with hemispherical hydrophobic attraction and found key differences from those characteristic of molecular amphiphiles. Experimental visualization combined with theory and molecular dynamics simulation shows that small, kinetically favored isomers fuse, before they equilibrate, into fibrillar triple helices with at most six nearest neighbors per particle. The time scales of colloidal rearrangement combined with the directional interactions resulting from Janus geometry make this a prototypical system to elucidate, on a mechanistic level and with single-particle kinetic resolution, how chemical anisotropy and reaction kinetics coordinate to generate highly ordered structures.

Confining potential when a biopolymer filament reptates.

Using single-molecule fluorescence imaging, we track Brownian motion perpendicular to the contour of tightly entangled F-actin filaments and extract the confining potential. The chain localization presents a small-displacement Hookean regime followed by a large amplitude regime where the effective restoring force is independent of displacement. The implied heterogeneity characterized by a distribution of tube width is modeled.