Simple synthesis of soft, tough, and cytocompatible biohybrid composites.

Collagen is the most abundant component of mammalian extracellular matrices. As such, the development of materials that mimic the biological and mechanical properties of collagenous tissues is an enduring goal of the biomaterials community. Despite the development of molded and 3D printed collagen hydrogel platforms, their use as biomaterials and tissue engineering scaffolds is hindered by either low stiffness and toughness or processing complexity. Here, we demonstrate the development of stiff and tough biohybrid composites by combining collagen with a zwitterionic hydrogel through simple mixing. This combination led to the self-assembly of a nanostructured fibrillar network of collagen that was ionically linked to the surrounding zwitterionic hydrogel matrix, leading to a composite microstructure reminiscent of soft biological tissues. The addition of 5-15 mg mL-1 collagen and the formation of nanostructured fibrils increased the elastic modulus of the composite system by 40% compared to the base zwitterionic matrix. Most notably, the addition of collagen increased the fracture energy nearly 11-fold ([Formula: see text] 180 J m-2) and clearly delayed crack initiation and propagation. These composites exhibit elastic modulus ([Formula: see text] 0.180 MJ) and toughness ([Formula: see text]0.617 MJ m-3) approaching that of biological tissues such as articular cartilage. Maintenance of the fibrillar structure of collagen also greatly enhanced cytocompatibility, improving cell adhesion more than 100-fold with >90% cell viability.

A novel non­selective atypical PKC agonist could protect neuronal cell line from Aß­oligomer induced toxicity by suppressing Aß generation.

Atypical protein kinase C (aPKCs) serve key functions in embryonic development by regulating apical­basal polarity. Previous studies have shed light on their roles during adulthood, especially in the development of Alzheimer's disease (AD). Although the crystal structure of PKCι has been resolved, an agonist of aPKCs remains to be discovered. In the present study, by using the Discovery Studio program and LibDock methodology, a small molecule library (K66­X4436 KINA Set) of compounds were screened for potential binding to PKCι. Subsequently, the computational docking results were validated using affinity selection­mass spectrometry, before in vitro kinase activity was used to determine the function of the hit compounds. A cell­based model assay that can mimic the pathology of AD was then established and used to assess the function of these hit compounds. As a result, the aPKC agonist Z640 was identified, which could bind to PKCι in silico, in vitro and in this cell­based model. Z640 was further confirmed as a non­selective aPKC agonist that can activate the kinase activity of both PKCι and PKCζ. In the cell­based assay, Z640 was found to protect neuronal cell lines from amyloid­ß (Aß) oligomer­induced cell death by reducing reactive oxygen species production and restore mitochondrial function. In addition, Z640 could reduce Aß40 generation in a dose­dependent manner and shift amyloid precursor protein processing towards the non­amyloid pathway. To conclude, the present study is the first, to the best of the authors' knowledge to identify an aPKC agonist by combining computer­assisted drug discovery and cell­based assays. The present study also revealed that aPKC agonists have therapeutic potential for the treatment of AD.

Acoustophoretic Liquefaction for 3D Printing Ultrahigh-Viscosity Nanoparticle Suspensions.

An acoustic liquefaction approach to enhance the flow of yield stress fluids during Digital Light Processing (DLP)-based 3D printing is reported. This enhanced flow enables processing of ultrahigh-viscosity resins (µapp  > 3700 Pa s at shear rates γ ˙  = 0.01 s-1 ) based on silica particles in a silicone photopolymer. Numerical simulations of the acousto-mechanical coupling in the DLP resin feed system at different agitation frequencies predict local resin flow velocities exceeding 100 mm s-1 at acoustic transduction frequencies of 110 s-1 . Under these conditions, highly loaded particle suspensions (weight fractions, ϕ = 0.23) can be printed successfully in complex geometries. Such mechanically reinforced composites possess a tensile toughness 2000% greater than the neat photopolymer. Beyond an increase in processible viscosities, acoustophoretic liquefaction DLP (AL-DLP) creates a transient reduction in apparent viscosity that promotes resin recirculation and decreases viscous adhesion. As a result, acoustophoretic liquefaction Digital Light Processing (AL-DLP) improves the printed feature resolution by more than 25%, increases printable object sizes by over 50 times, and can build parts >3 × faster when compared to conventional methodologies.

Making bioinspired 3D-printed autonomic perspiring hydrogel actuators.

To mitigate the adverse effects of elevated temperatures, conventional rigid devices use bulky radiators, heat sinks and fans to dissipate heat from sensitive components. Unfortunately, these thermoregulation strategies are incompatible with soft robots, a growing field of technology that, like biology, builds compliant and highly deformable bodies from soft materials to enable functional adaptability. Here, we design fluidic elastomer actuators that autonomically perspire at elevated temperatures. This strategy incurs operational penalties (i.e., decreased actuation efficiency and loss of hydraulic fluid) but provides for thermoregulation in soft systems. In this bioinspired approach, we 3D-print finger-like actuators from smart gels with embedded micropores that autonomically dilate and contract in response to temperature. During high-temperature operation, the internal hydraulic fluid flows through the dilated pores, absorbs heat and vaporizes. Upon cooling, the pores contract to restrict fluid loss and restore operation. To assess the thermoregulatory performance, this protocol uses non-invasive thermography to measure the local temperatures of the robot under varied conditions. A mathematical model based on Newton's law of cooling quantifies the cooling performance and enables comparison between competing designs. Fabrication of the sweating actuator usually takes 3-6 h, depending on size, and can provide >100 W/kg of additional cooling capacity.

Autonomic perspiration in 3D-printed hydrogel actuators.

In both biological and engineered systems, functioning at peak power output for prolonged periods of time requires thermoregulation. Here, we report a soft hydrogel-based actuator that can maintain stable body temperatures via autonomic perspiration. Using multimaterial stereolithography, we three-dimensionally print finger-like fluidic elastomer actuators having a poly-N-isopropylacrylamide (PNIPAm) body capped with a microporous (~200 micrometers) polyacrylamide (PAAm) dorsal layer. The chemomechanical response of these hydrogel materials is such that, at low temperatures (<30°C), the pores are sufficiently closed to allow for pressurization and actuation, whereas at elevated temperatures (>30°C), the pores dilate to enable localized perspiration in the hydraulic actuator. Such sweating actuators exhibit a 600% enhancement in cooling rate (i.e., 39.1°C minute-1) over similar non-sweating devices. Combining multiple finger actuators into a single device yields soft robotic grippers capable of both mechanically and thermally manipulating various heated objects. The measured thermoregulatory performance of these sweating actuators (~107 watts kilogram-1) greatly exceeds the evaporative cooling capacity found in the best animal systems (~35 watts kilogram-1) at the cost of a temporary decrease in actuation efficiency.

3D printable tough silicone double networks.

Additive manufacturing permits innovative soft device architectures with micron resolution. The processing requirements, however, restrict the available materials, and joining chemically dissimilar components remains a challenge. Here we report silicone double networks (SilDNs) that participate in orthogonal crosslinking mechanisms-photocurable thiol-ene reactions and condensation reactions-to exercise independent control over both the shape forming process (3D printing) and final mechanical properties. SilDNs simultaneously possess low elastic modulus (E100% < 700kPa) as well as large ultimate strains (dL/L0 up to ~ 400 %), toughnesses (U ~ 1.4 MJ·m-3), and strengths (σ ~ 1 MPa). Importantly, the latent condensation reaction permits cohesive bonding of printed objects to dissimilar substrates with modulus gradients that span more than seven orders of magnitude. We demonstrate soft devices relevant to a broad range of disciplines: models that simulate the geometries and mechanical properties of soft tissue systems and multimaterial assemblies for next generation wearable devices and robotics.

3D Printing of Viscoelastic Suspensions via Digital Light Synthesis for Tough Nanoparticle-Elastomer Composites.

The rheological parameters required to print viscoelastic nanoparticle suspensions toward tough elastomers via Digital Light Synthesis (DLS) (an inverted projection stereolithography system) are reported. With a model material of functionalized silica nanoparticles suspended in a poly(dimethylsiloxane) matrix, the rheological-parameters-guided DLS can print structures seven times tougher than those formed from the neat polymers. The large yield stress and high viscosity associated with these high concentration nanoparticle suspensions, however, may prevent pressure-driven flow, a mechanism essential to stereolithography-based printing. Thus, to better predict and evaluate the printability of high concentration nanoparticle suspensions, the boundary of rheological properties compatible with DLS is defined using a non-dimensional Peclet number (Pe). Based on the proposed analysis of rheological parameters, the border of printability at standard temperature and pressure (STP) is established by resin with a silica nanoparticle mass fraction (ϕsilica ) of 0.15. Above this concentration, nanoparticle suspensions have Pe > 1 and are not printable. Beyond STP, the printability can be further extended to ϕsilica = 0.20 via a heating module with lower shear rate to reduce the Pe < 1. The printed rubber possesses even higher toughness (Γ ≈ 155 kJ m-3 ), which is 40% higher over that of ϕsilica = 0.15.

Scalable Synthesis of Switchable Assemblies of Gold Nanorod Lyotropic Liquid Crystal Nanocomposites.

A new class of solvent free, lyotropic liquid crystal nanocomposites based on gold nanorods (AuNRs) with high nanorod content is reported. Application of shear results in switchable, highly ordered alignment of the nanorods over several centimeters with excellent storage stability for months. For the synthesis, AuNRs are surface functionalized with a charged, covalently tethered corona, which induces fluid-like properties. This honey-like material can be deposited on a substrate and a high orientational order parameter of 0.72 is achieved using a simple shearing protocol. Switching shearing direction results in realignment of the AuNRs. For a film containing 75 wt% of AuNRs the alignment persists for several months. In addition to the lyotropic liquid crystal characteristics, the AuNRs films also exhibit anisotropic electrical conductivity with an order of magnitude difference between the conductivities in direction parallel and perpendicular to the alignment of the AuNRs.

Simple Synthesis of Elastomeric Photomechanical Switches That Self-Heal.

This article introduces a simple two-stage method to synthesize and program a photomechanical elastomer (PME) for light-driven artificial muscle-like actuations in soft robotics. First, photochromic azobenzene molecules are covalently attached to a polyurethane backbone via a two-part step-growth polymerization. Next, mechanical alignment is applied to induce anisotropic deformations in the PME-actuating films. Cross-linked through dynamic hydrogen bonds, the PMEs also possess autonomic self-healing properties without external energy input. This self-healing allows for a single alignment step of the PME film and subsequent "cut and paste" assembly for multi-axis actuation of a self-folded soft-robotic gripper from a single degree of freedom optical input.

Optical stereolithography of antifouling zwitterionic hydrogels.

This paper reports the rapid 3D printing of tough (toughness, UT, up to 141.6 kJ m-3), highly solvated (φwater∼ 60 v/o), and antifouling hybrid hydrogels for potential uses in biomedical, smart materials, and sensor applications, using a zwitterionic photochemistry compatible with stereolithography (SLA). A Design of Experiments (DOE) framework was used for systematically investigating the multivariate photochemistry of SLA generally and, specifically, to determine an aqueous SLA system with an additional zwitterionic acrylate, which significantly increases the gelation rate, and the resilience of the resulting hybrid hydrogels relative to an equivalent non-ionic polyacrylamide hydrogel. Specifically, the resulting zwitterionic hybrid hydrogels (Z-gels) can be tuned over a large range of ultimate strains, ca. 0.5 < γult < 5.0, and elastic moduli, ca. 10 < E < 1000 kPa, while also demonstrating a high resilience under cyclic tensile loading. Importantly, unlike traditional chemistry, increasing the elastic modulus of the Z-gels does not necessarily reduce the ultimate strain. Moreover, the Z-gels can be rapidly printed using a desktop commercial SLA 3D printer, with relatively low photoirradiation dosages of visible light (135 to 675 mJ cm-2 per 50-100 µm layer). Compared with the counterpart polyacrylamide hydrogels, the Z-gels have greater antifouling properties and exhibit 58.2% less absorption of bovine serum albumin.

Solution-printed organic semiconductor blends exhibiting transport properties on par with single crystals.

Solution-printed organic semiconductors have emerged in recent years as promising contenders for roll-to-roll manufacturing of electronic and optoelectronic circuits. The stringent performance requirements for organic thin-film transistors (OTFTs) in terms of carrier mobility, switching speed, turn-on voltage and uniformity over large areas require performance currently achieved by organic single-crystal devices, but these suffer from scale-up challenges. Here we present a new method based on blade coating of a blend of conjugated small molecules and amorphous insulating polymers to produce OTFTs with consistently excellent performance characteristics (carrier mobility as high as 6.7 cm(2) V(-1) s(-1), low threshold voltages of<1 V and low subthreshold swings <0.5 V dec(-1)). Our findings demonstrate that careful control over phase separation and crystallization can yield solution-printed polycrystalline organic semiconductor films with transport properties and other figures of merit on par with their single-crystal counterparts.

Estimation of inhibitory effects of hemicellulosic wood hydrolysate inhibitors on PHA production by Burkholderia cepacia ATCC 17759 using response surface methodology.

Sugar maple hemicellulosic hydrolysate was utilized as a renewable feedstock for polyhydroxyalkanoates production by Burkholderia cepacia ATCC 17759. To estimate inhibitory effects of the hydrolysate, response surface methodology was utilized to analyze cell growth and PHA accumulation in the presence of multiple inhibitors. Mixture design was employed to study the correlation between the proportion of phenolics and total inhibition. The resultant models (R(2) as 92.42% and 93.14% for cell growth and PHA production, respectively) indicated syringic acid was the most inhibitory among three phenolics and synergistic inhibition was observed for the combinations of vanillin/syringic acid and vanillic acid/syringic acid. When furfural, levulinic acid, and acetate were also present during the fermentation, central composite design was employed. The regression model using 48 h cell growth as the response surface (R(2)=87.82%) showed acetate was the most inhibitory. Additionally, strong synergistic effects were observed for the combinations of acetate/phenolics and levulinic acid/furfural.

Production of polyhydroxyalkanoates by Burkholderia cepacia ATCC 17759 using a detoxified sugar maple hemicellulosic hydrolysate.

Sugar maple hemicellulosic hydrolysate containing 71.9 g/l of xylose was used as an inexpensive feedstock to produce polyhydroxyalkanoates (PHAs) by Burkholderia cepacia ATCC 17759. Several inhibitory compounds present in wood hydrolysate were analyzed for effects on cell growth and PHA production with strong inhibition observed at concentrations of 1 g/l furfural, 2 g/l vanillin, 7 g/l levulinic acid, and 1 M acetic acid. Gradual catabolism of lower concentrations of these inhibitors was observed in this study. To increase the fermentability of wood hydrolysate, several detoxification methods were tested. Overliming combined with low-temperature sterilization resulted in the highest removal of total inhibitory phenolics (65%). A fed-batch fermentation exhibited maximum PHA production after 96 h (8.72 g PHA/L broth and 51.4% of dry cell weight). Compositional analysis by NMR and physical-chemical characterization showed that PHA produced from wood hydrolysate was composed of polyhydroxybutyrate (PHB) with a molecular mass (M (N)) of 450.8 kDa, a melting temperature (T (m)) of 174.4°C, a glass transition temperature (T (g)) of 7.31°C, and a decomposition temperature (T (decomp)) of 268.6°C.

Toxicity of pharmaceutical wastewater on male reproductive system of Mus musculus.

This study reports on the toxic effects of 35-days intragastric perfusion of pharmaceutical wastewater on the male reproductive system of Mus musculus. Flow cytometric analyses and staining with fluorescein diacetate (FDA) and propidium iodide (PI) were used to assess the toxicity on spermatogenic cells. Significant depletions in the relative percentages of elongated spermatid (HC), diploid spermatogonia (2C), and S-phase cells were observed. These alterations in different germ cell populations were reflected in the various germ cell ratios. The ratios of 1C:4C and HC:2C showed a significant decline after pharmaceutical wastewater treatment, while the 4C:2C and 1C:2C ratios increased significantly. FDA and PI staining displayed reduced viability of spermatogenic cells in wastewater treated group. Statistically significant percentages of sperm abnormalities showed the genotoxic potential of this pharmaceutical wastewater. Testicular histopathological studies of treated animals revealed expansion of interstitial space and reduction in the number and size of Leydig cells. Thus, the present study has established the toxicity of pharmaceutical wastewater on the reproductive biology of male mice.

PTA wastewater molecular toxicity detected with gene chip.

The purified terephthalic acid (PTA) petrochemical wastewater molecular toxicity detected by use of Mouse Genome 430A 2.0 GeneChip was conducted in this research. The toxic dose to male mice was 0.03 g/(kg x d) of PTA in the wastewater. The mice liver total RNA was isolated as the temple for synthesis of cDNA and then the cDNA as the temple for synthesis of cRNA. Hybridizing the cRNA with the target genes on the gene chip, there were 232 genes expression levels up-regulated and 74 genes down-regulated discovered obviously. The foremost 40 genes for both the highest and the lowest expression levels involved endogenetic steroid and hormone metabolism, immune system, the leukocyte activity and inflammation, detoxification in liver, reproduction and growth hormone, regulation immune factors of anti-tumor and anti-infection and cancer to the mice sampled. The data suggest the PTA wastewater contained over 5 aromatics and their toxicities integrated were much higher than the pure chemical PTA. And the pure chemical PTA toxicities data cannot be used to evaluate the toxicity of the PTA wastewater instead.