The mechano-chemical circuit drives skin organoid self-organization.

Stem cells in organoids self-organize into tissue patterns with unknown mechanisms. Here, we use skin organoids to analyze this process. Cell behavior videos show that the morphological transformation from multiple spheroidal units with morphogenesis competence (CMU) to planar skin is characterized by two abrupt cell motility-increasing events before calming down. The self-organizing processes are controlled by a morphogenetic module composed of molecular sensors, modulators, and executers. Increasing dermal stiffness provides the initial driving force (driver) which activates Yap1 (sensor) in epidermal cysts. Notch signaling (modulator 1) in epidermal cyst tunes the threshold of Yap1 activation. Activated Yap1 induces Wnts and MMPs (epidermal executers) in basal cells to facilitate cellular flows, allowing epidermal cells to protrude out from the CMU. Dermal cell-expressed Rock (dermal executer) generates a stiff force bridge between two CMU and accelerates tissue mixing via activating Laminin and ß1-integrin. Thus, this self-organizing coalescence process is controlled by a mechano-chemical circuit. Beyond skin, self-organization in organoids may use similar mechano-chemical circuit structures.

No transmission of porcine endogenous retrovirus in an acute liver failure model treated by a novel hybrid bioartificial liver containing porcine hepatocytes.

BACKGROUND: A novel hybrid bioartificial liver (HBAL) was constructed using an anionic resin adsorption column and a multi-layer flat-plate bioreactor containing porcine hepatocytes co-cultured with bone marrow mesenchymal stem cells (MSCs). This study aimed to evaluate the microbiological safety of the HBAL by detecting the transmission of porcine endogenous retroviruses (PERVs) into canines with acute liver failure (ALF) undergoing HBAL. METHODS: Eight dogs with ALF received a 6-hour HBAL treatment on the first day after the modeling by D-galactosamine administration. The plasma in the HBAL and the whole blood in the dogs were collected for PERV detection at regular intervals until one year later when the dogs were sacrificed to retrieve the tissues of several organs for immunohistochemistry and Western blotting for the investigation of PERV capsid protein gag p30 in the tissue. Furthermore, HEK293 cells were incubated to determine the in vitro infectivity. RESULTS: PERV RNA and reverse transcriptase activity were observed in the plasma of circuit 3, suggesting that PERV particles released in circuit 3. No positive PERV RNA and reverse transcriptase activity were detected in other plasma. No HEK293 cells were infected by the plasma in vitro. In addition, all PERV-related analyses in peripheral blood mononuclear cells and tissues were negative. CONCLUSION: No transmission of PERVs into ALF canines suggested a reliable microbiological safety of HBAL based on porcine hepatocytes.

New strategy for surface functionalization of periodic mesoporous silica based on meso-HSiO1.5.

Organic functionalization of periodic mesoporous silicas (PMSs) offers a way to improve their excellent properties and wide applications owing to their structural superiority. In this study, a new strategy for organic functionalization of PMSs is demonstrated by hydrosilylation of the recently discovered "impossible" periodic mesoporous hydridosilica, meso-HSiO1.5. This method overcomes the disadvantages of present pathways for organic functionalization of PMSs with organosilica. Moreover, compared to the traditional functionalization on the surface of porous silicon by hydrosilylation, the template-synthesized meso-HSiO1.5 is more flexible to access functional-groups-loaded PMSs with adjustable microstructures. The new method and materials will have wider applications based on both the structure and surface superiorities.

The mechano-chemical circuit in fibroblasts and dendritic cells drives basal cell proliferation in psoriasis.

Psoriasis is an intractable immune-mediated disorder that disrupts the skin barrier. While studies have dissected the mechanism by which immune cells directly regulate epidermal cell proliferation, the involvement of dermal fibroblasts in the progression of psoriasis remains unclear. Here, we identified that signals from dendritic cells (DCs) that migrate to the dermal-epidermal junction region enhance dermal stiffness by increasing extracellular matrix (ECM) expression, which further promotes basal epidermal cell hyperproliferation. We analyzed cell-cell interactions and observed stronger interactions between DCs and fibroblasts than between DCs and epidermal cells. Using single-cell RNA (scRNA) sequencing, spatial transcriptomics, immunostaining, and stiffness measurement, we found that DC-secreted LGALS9 can be received by CD44+ dermal fibroblasts, leading to increased ECM expression that creates a stiffer dermal environment. By employing mouse psoriasis and skin organoid models, we discovered a mechano-chemical signaling pathway that originates from DCs, extends to dermal fibroblasts, and ultimately enhances basal cell proliferation in psoriatic skin.

Highly Aligned Poly(3,4-ethylene dioxythiophene) (PEDOT) Nano- and Microscale Fibers and Tubes.

This study reports a facile method for the fabrication of aligned Poly(3,4-ethylene dioxythiophene) (PEDOT) fibers and tubes based on electrospinning and oxidative chemical polymerization. Discrete PEDOT nano- and microfibers and nano- and microtubes are difficult to fabricate quickly and reproducibly. We employed poly(lactide-co-glycolide) (PLGA) polymers that were loaded with polymerizable 3,4-ethylene dioxythiophene (EDOT) monomer to create aligned nanofiber assemblies using a rotating glass mandrel during electrospinning. The EDOT monomer/PLGA polymer blends were then polymerized by exposure to an oxidative catalyst (FeCl3). PEDOT was polymerized by continuously dripping a FeCl3 solution onto the glass rod during electrospinning. The resulting PEDOT fibers were conductive, aligned and discrete. Fiber bundles could be easily produced in lengths of several centimeters. The PEDOT sheath/PLGA core fibers were immersed in chloroform to remove the PLGA and any residual EDOT resulting in hollow PEDOT tubes. This approach made it possible to easily generate large areas of aligned PEDOT fibers/tubes. The structure and properties of the aligned assemblies were measured using optical microscopy, electron microscopy, Raman spectroscopy, thermal gravimetric analysis, and DC conductivity measurements. We also demonstrated that the aligned PEDOT sheath/PLGA core fiber assemblies could be used in supporting and directing the extension of dorsal root ganglia (DRG) neurons in vitro.

A Skin-Inspired Multifunctional Conductive Hydrogel with High Stretchable, Adhesive, Healable, and Decomposable Properties for Highly Sensitive Dual-Sensing of Temperature and Strain.

Developing smart hydrogels with excellent physicochemical properties and multi-sensing capabilities for various simulation of human skin's functions still remains a great challenge. Here, based on simple and convenient one-step covalent cross-linking method enhanced by dynamic RS-Ag interactions, a skin-inspired multifunctional conductive hydrogel with desirable physicochemical properties (including high stretchability, self-adhesion, self-healing, decomposition and removability) is developed for highly sensitive dual-sensing of temperature and strain. Benefiting from the synergistic action of multiple hydrogen bonds, RS-Ag bonds and S-S bonds, the gel exhibited a novel thermosensitive mechanism. The prepared hydrogels exhibited extremely high mechanical properties (maximum tensile strength of 0.35 MPa, elongation at break nearly 1800%, compressive stress over 4.43 MPa), excellent self-healing (96.82% (stress), 88.45% (temperature), 73.89% (mechanical property)), decomposition (the molecular weight after decomposition is below 700) and self-adhesion (enhanced contact with the material interface). In addition, this conductive hydrogel could also simultaneously achieve highly sensitive temperature-sensing (TCR: 10.89) and stress-sensing (GF: 1.469). As a proof-to-concept, the hydrogel displayed superior capability for simulation of human skin to perception of touch, pressure and ambient temperature simultaneously, indicating promising applications in the fields of wearable devices, personal health care, and human-machine interfaces.

Super-Long SERS Active Single Silver Nanowires for Molecular Imaging in 2D and 3D Cell Culture Models.

Establishing a systematic molecular information analysis strategy for cell culture models is of great significance for drug development and tissue engineering technologies. Here, we fabricated single silver nanowires with high surface-enhanced Raman scattering activity to extract SERS spectra in situ from two-dimensional (2D) and three-dimensional (3D) cell culture models. The silver nanowires were super long, flexible and thin enough to penetrate through multiple cells. A single silver nanowire was used in combination with a four-dimensional microcontroller as a cell endoscope for spectrally analyzing the components in cell culture models. Then, we adopted a machine learning algorithm to analyze the obtained spectra. Our results show that the abundance of proteins differs significantly between the 2D and 3D models, and that nucleic acid-rich and protein-rich regions can be distinguished with satisfactory accuracy.

Periodic mesoporous hydridosilica--synthesis of an "impossible" material and its thermal transformation into brightly photoluminescent periodic mesoporous nanocrystal silicon-silica composite.

There has always been a fascination with "impossible" compounds, ones that do not break any rules of chemical bonding or valence but whose structures are unstable and do not exist. This instability can usually be rationalized in terms of chemical or physical restrictions associated with valence electron shells, multiple bonding, oxidation states, catenation, and the inert pair effect. In the pursuit of these "impossible" materials, appropriate conditions have sometimes been found to overcome these instabilities and synthesize missing compounds, yet for others these tricks have yet to be uncovered and the materials remain elusive. In the scientifically and technologically important field of periodic mesoporous silicas (PMS), one such "impossible" material is periodic mesoporous hydridosilica (meso-HSiO(1.5)). It is the archetype of a completely interrupted silica open framework material: its pore walls are comprised of a three-connected three-dimensional network that should be so thermodynamically unstable that any mesopores present would immediately collapse upon removal of the mesopore template. In this study we show that meso-HSiO(1.5) can be synthesized by template-directed self-assembly of HSi(OEt)(3) under aqueous acid-catalyzed conditions and after template extraction remains stable to 300 °C. Above this temperature, bond redistribution reactions initiate a metamorphic transformation which eventually yields periodic mesoporous nanocrystalline silicon-silica, meso-ncSi/SiO(2), a nanocomposite material in which brightly photoluminescent silicon nanocrystallites are embedded within a silica matrix throughout the mesostructure. The integration of the properties of silicon nanocrystallinity with silica mesoporosity provides a wealth of new opportunities for emerging nanotechnologies.

The study of adsorption characteristics of electrospun polymer nanofibers for benzenes in water.

The adsorption properties of benzene, p-dichlorobenzene and nitrobenzene on polymer nanofibers were studied. Compared with polyacrylonitrile nanofiber, polystyrene (PS) nanofiber presented better adsorption performance. Langmuir and Freundlich adsorption models were used for the mathematical description of adsorption equilibria, and Freundlich isotherms fitted better. Kinetic studies showed that the adsorption of PS nanofiber followed pseudo first-order model. Various thermodynamic parameters such as standard free energy (delta G), enthalpy (delta H) and entropy (delta S) were calculated for predicting the adsorption nature of PS nanofiber for three benzenes, which indicated that the adsorption was spontaneous and a physical process. The regeneration efficiency maintains over 80% after five cycles of adsorption/desorption tests. It showed that PS nanofibers are promising candidates for adsorption and removal of aromatic hydrocarbons from water.

Structural color films with lotus effects, superhydrophilicity, and tunable stop-bands.

The structural blue color of a Morpho butterfly originates from the diffraction of light and interference effects due to the presence of the microstructures on the wing of the butterfly. Structural color on the surface of a damselfish reversibly changes between green and blue. Inspired by these creatures, we have been trying to prepare high-quality and functional structural color films. We describe our efforts in this Account. A useful technique to prepare such structural color films in colloidal solution is a "lifting" method, which allows us to quickly fabricate brilliant colloidal crystal films. The thicknesses of the films can be controlled by precisely adjusting the particle concentration and the lifting speed. Moreover, in order to prepare a complicated structure, we have used template methods. Indeed, we have successfully prepared the inverse structure of the wing of a Morpho butterfly with this technique. Initially, however, our structural color films had a whitish appearance due to the scattering of light by defects in the colloidal crystal film. Later, we were able to prepare a non-whitish structural color film by doping an appropriate dye in the colloidal particles to absorb the scattering light. In addition to the structural blue color, the wing of the Morpho butterfly has superhydrophobic properties. According to Wenzel's equation, the hydrophobic and hydrophilic properties are enhanced when the roughness of the hydrophobic and hydrophilic surface is increased, respectively. Based on this mechanism, we have successfully prepared structural color films with superhydrophobic properties, as well as with superhydrophilic properties. Another important property that can be seen in nature is tunable structural color, such as the color change that can be seen on the surface of a damselfish. In order to mimic such color change, we have developed several tunable structural color films. In particular, we have successfully prepared phototunable photonic crystals using photoresponsive azobenzene derivatives. In order to apply these structural color films, we developed a technique for patterning them by taking advantage of the wettability of the substrate surface. These materials can be used in the future for self-cleaning pigments and tunable photonic crystals.

Application of nanofiber-packed SPE for determination of salivary-free cortisol using fluorescence precolumn derivatization and HPLC detection.

Salivary cortisol has emerged as an easy-to-collect biologic marker of stress in many researches. In this study, we present a method for the determination of salivary-free cortisol using HPLC method with fluorescence precolumn derivatization, which is based on a novel extraction from the strongly acidic medium (fluorescent derivatives of cortisol in sulfuric acid medium) by electrospun polystyrene nanofibers packed SPE. For high-throughput sample extraction, an array pretreatment device based on nanofibers packed SPE micro-column was designed. The LOD of cortisol was 0.01 microg/L (S/N=3). The RSDs (n=6) for all analytes were below 8.0%, and the recoveries were 110, 102.4, and 99.4% (n=3) for saliva spiked with 0.1, 10, and 20 microg/L of cortisol, respectively. The proposed method was then successfully applied in the determination of free cortisol in human saliva. The salivary cortisol concentrations in the real samples ranged from 0.22 to 7.45 microg/L. The nanofiber-packed SPE overcame the low extraction recovery and bad clean-up effect of the conventional methods, and increased the sensitivity and selectivity of the method.

Gel-pad microarrays templated by patterned porous silicon for dual-mode detection of proteins.

A proof-of-concept study demonstrated the feasibility of a novel gel-pad microarray on porous silicon chips, by initiation of an atom transfer radical propagation (ATRP) polymerisation of (polyethylene glycol) methacrylate (PEGMA) with surface Si-H species, stepwise chemical conversions of the gel membrane to an NTA-Ni2+/histidine-tagged protein system, and matrix-assisted laser desorption/ionisation mass spectroscopy (MALDI MS) and fluorescence detections.

Design of packed-fiber solid-phase extraction device for analysis of the drug and its metabolite in plasma.

A mini-column packed with 1 mg electrospun polystyrene nanofibers (about 200 approximately 400 nm in diameter) was designed for simple, fast extraction of drugs, diazepam and its major metabolite, N-desmethyldiazepam for the analysis of them in human and dog plasma. Ttrezodone was selected as internal standard. The drugs adsorbed on the solid phase could be desorpted with 50 microl of the methanol and then monitored by liquid chromatography coupled to an ultraviolet detector. Parameters influencing the extraction efficiency such as fiber packing amount, eluted solvent, and pH of the sample were decided. The time for the pretreatment of 0.5 ml plasma sample was less than 10 min. The detection limits of diazepam and N-desmethyldiazepam in plasma could be as low as 1 microg/L. The intra- and inter-day precision, calculated from quality control (QC) samples, was less than 9.1%. The method was evaluated by its application in determination of dog plasma samples from three beagles after a single dose oral of diazepam. The technique was validated by comparison with conventional plasma analysis. It was observed that the mini-column offers improved limits of detection and reduced sample preparation time as compared to conventional method. For its simplicity and sensitivity, the method may be used in therapeutic drug monitoring and pharmacology study.

Packing the silica colloidal crystal beads: a facile route to superhydrophobic surfaces.

To mimic the structure of the lotus leaf, we present a facile route to prepare superhydrophobic surfaces by depositing nanoparticle clusters onto a solid surface. These clusters were fabricated via solidification of an emulsion droplet containing a nanoparticle in silicone oil. Thus, the microsized clusters and nanoparticles form dual-scale roughness structures. The surface is modified by fluoroalkylsilane and exhibits superhydrophobicity, with a contact angle greater than 165 degrees as well as a sliding angle less than 1 degrees . On the basis of size tuning of the nano/microstructures, various contact angles and sliding angles were investigated. Furthermore, the influence of micro/nanostructures on superhydrophobicity is discussed.

Quantum-dot-coated encoded silica colloidal crystals beads for multiplex coding.

Multiplex optical coding carriers for biological assays have been achieved by coating silica colloidal crystals beads with quantum dots, which provided high stability, large capacity and simplicity for practical application.

Boron-doped diamond nanograss array for electrochemical sensors.

A novel BDD nanograss array has been prepared simply on a heavily doped BDD film by reactive ion etching for use as an electrochemical sensor, which improved the reactive site, promoted the electrocatalytic activity, accelerated the electron transfer, and enhanced the selectivity.

Chitosan nanofiber scaffold enhances hepatocyte adhesion and function.

To enhance cell attachment and promote liver functions of hepatocytes cultured in bioreactors, a chitosan nanofiber scaffold was designed and prepared via electrospinning. Effects of the scaffold on hepatocyte adhesion, viability and function were then investigated. Data showed that hepatocytes on chitosan nanofiber scaffold exhibited better viability and tighter cell-substrate contact than cells on regular chitosan film. In addition, urea synthesis, albumin secretion and cytochrome P450 activity of hepatocytes on chitosan nanofiber scaffold were all 1.5 to 2 folds higher than the controls. Glycogen synthesis was also increased as compared with the controls. These results suggested the potential application of this chitosan nanofiber scaffold as a suitable substratum for hepatocyte culturing in bioreactors.

Blu-Ray Discs as Universal Biochip Substrates: Lithography-Free Surface Activation and Assay Patterning.

Blu-ray discs (BDs) are advantageous in comparison with other optical discs (compact discs and digital versatile discs) in terms of not only their storage capacity but also the high-quality materials fabricated from. We have recently discovered that the "Hard Coat" film of Verbatim BDs is in fact a unique type of polymeric substrates that can be readily activated and adapted for biochip fabrications. Particularly, the Hard Coat film peeled from BDs is optically transparent without any fluorescence background, which can be activated by treating with a common base (1.0 M NaOH) at a slightly elevated temperature (55 °C). The surface density of reactive carboxylic acid groups generated, 6.6 ± 0.7 × 10-9 mol/cm2, is much higher than that on polycarbonate upon UV/ozone irradiation (4.8 ± 0.2 × 10-10 mol/cm2). There are no significant physical damages to the substrate morphology, and the aging effect is minimal. More importantly, the BD substrate can be patterned using either cut-out filter paper masks or microfluidic channel plates; both are lithography-free, bench-top methods that facilitate the device fabrication in a common laboratory setting. With classical biotin-streptavidin binding and DNA hybridization arrays as trial systems, we have also demonstrated this new type of biochip substrates for quantitative assay applications.

Fiber-packed SPE tips based on electrospun fibers.

A novel fiber-packed solid-phase extraction (SPE) tip was designed based on electrospun nanofibers. The tip was used to investigate the extraction of hydrocortisone (HC), cortisone acetate (CA), ethinylestradiol (EE), and estradiol (E2). The effects of diameters, porous figurations, and functional groups of the electrospun fibers on the selectivity and efficiency were studied. The experimental results indicated that the detection limit of cortisol in water sample could be as low as 0.75 ng/mL. When the tip is used for detection of cortisol in human hair the efficiency of biological sample pretreatment is better than the traditional SPE method. Our method could significantly simplify the traditional SPE process and lower the cost. Industrial application of the tip is anticipated.

Performance of electrospun nanofibers for SPE of drugs from aqueous solutions.

A novel extraction technique was reported. The solid phase material, nanofiber, was prepared by electrospinning using polystyrene. Twenty different drugs (10 microg/L in water) were extracted using 1 mg of nanofibers within 5 min. The analytes can be desorpted from the fibers with 50 microL of the methanol and then monitored by LC coupled to a UV detector. Packed-fiber SPE (PFSPE) provide high recoveries (>50%) for some relatively non-polar drugs (log P >1.5) (n-octanol-to-water partition ratio), and relatively low recoveries (9.9-39.8%) for the drugs within the log P window below 1. Experimental optimization of the technique has been carried out using seven representative drugs, edaravone, cinchonine, quinine, voriconazole, chlordiazepoxide, verapamil, and rutonding. Except for edaravone, the maximum yields of seven drugs (0.2 microg/L) from water samples were approximately 100%, and were 33.7-88.2% from human plasma. The advantageous aspect of the technique encompasses high throughput, high sensitivity, simplicity, low cost, and green chemistry.