Aerosol Jet Printing-Enabled Dual-Function Electrochemical and Colorimetric Biosensor for SARS-CoV-2 Detection.

An aerosol jet printing-enabled dual-function biosensor for the sensitive detection of pathogens using SARS-CoV-2 RNA as an example has been developed. A CRISPR-Cas13:guide-RNA complex is activated in the presence of a target RNA, leading to the collateral trans-cleavage of ssRNA probes that contain a horseradish peroxidase (HRP) tag. This, in turn, catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by HRP, resulting in a color change and electrochemical signal change. The colorimetric and electrochemical sensing protocol does not require complicated target amplification and probe immobilization and exhibits a detection sensitivity in the femtomolar range. Additionally, our biosensor demonstrates a wide dynamic range of 5 orders of magnitude. This low-cost aerosol inkjet printing technique allows for an amplification-free and integrated dual-function biosensor platform, which operates at physiological temperature and is designed for simple, rapid, and accurate point-of-care (POC) diagnostics in either low-resource settings or hospitals.

Aerosol Jet Printing Enabled Dual-Function Electrochemical and Colorimetric Biosensor for SARS-CoV-2 Detection.

An aerosol jet printing enabled dual-function biosensor for the sensitive detection of pathogens using SARS-CoV-2 RNA as an example has been developed. A CRISPR-Cas13: guide-RNA complex is activated in the presence of a target RNA, leading to the collateral trans-cleavage of ssRNA probes that contain a horseradish peroxidase (HRP) tag. This, in turn, catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by HRP, resulting in a color change and electrochemical signal change. The colorimetric and electrochemical sensing protocol does not require complicated target amplification and probe immobilization and exhibits a detection sensitivity in the femtomolar range. Additionally, our biosensor demonstrates a wide dynamic range of 5 orders of magnitude. This low-cost aerosol inkjet printing technique allows for an amplification-free and integrated dual-function biosensor platform, which operates at physiological temperature and is designed for simple, rapid, and accurate point-of-care (POC) diagnostics in either low-resource settings or hospitals.

Gold Nanoparticle-Labeled CRISPR-Cas13a Assay for the Sensitive Solid-State Nanopore Molecular Counting.

A gold nanoparticle (AuNP) labeled CRISPR-Cas13a nucleic acid assay has been developed for sensitive solid-state nanopore sensing. Instead of directly detecting the translocation of RNA through a nanopore, our system utilizes non-covalent conjugates of AuNPs and RNA targets. Upon CRISPR activation, the AuNPs are liberated from the RNA, isolated, and passed through a nanopore sensor. Detection of the AuNPs can be observed as increasing ionic current in the chip. Each AuNP that is detected is enumerated as an event, leading to quantitative of molecular targets. Leveraging the high signal-to-noise ratio enabled by the AuNPs, a detection limit of 50 fM before front-end target amplification is achieved using SARS-CoV-2 RNA segments as a Cas13 target. Furthermore, a dynamic range of six orders of magnitude is demonstrated for quantitative RNA sensing. This simplified AuNP-based CRISPR assay is performed at the physiological temperature without relying on thermal cyclers. In addition, the nanopore reader is similar in size to a smartphone, making the assay system suitable for rapid and portable nucleic acid biomarker detection in either low-resource settings or hospitals.

Mechanical Response of Carbon Composite Octet Truss Structures Produced via Axial Lattice Extrusion.

Engineered lattice structures fabricated via additive manufacturing (AM) technologies are of great interest for many applications that require high strength and/or stiffness with minimum mass. This paper studies a novel axial lattice extrusion (ALE) AM technique that greatly enhances mechanical properties of polymeric lattice structures. When the novel ALE process was used to produce 84 mm × 84 mm × 84 mm octet truss lattice samples using fiber reinforced ABS, a total of 219,520 polymer interfaces in the lattice beams were eliminated relative to the conventional 3D printing alternative. Microscopic examination revealed near perfect alignment of the chopped carbon fibers with axes of the cylindrical beams that make up the lattice structure. The greatly enhanced beam quality with fiber reinforcement resulted in excellent mechanical properties. Compression testing yielded an average relative compressive strength of 17.4 MPa and an average modulus of 162.8 MPa. These properties rate very strongly relative to other published work, and indicate that the ALE process shows great potential for fabrication of high-strength, lightweight, large-scale, carbon-fiber composite components. The paper also contributes a modeling approach to finite element analysis (FEA) that captures the highly orthotropic properties of carbon fiber lattice beams. The diagonal shear failure mode predicted via the FEA model was in good agreement with experimentally observed results.

Magnetic Field Patterning of Nickel Nanowire Film Realized by Printed Precursor Inks.

This paper demonstrates an easily prepared novel material and approach to producing aligned nickel (Ni) nanowires having unique and customizable structures on a variety of substrates for electronic and magnetic applications. This is a new approach to producing printed metallic Ni structures from precursor materials, and it provides a novel technique for nanowire formation during reduction. This homogeneous solution can be printed in ambient conditions, and it forms aligned elemental Ni nanowires over large areas upon heating in the presence of a magnetic field. The use of templates or subsequent purification are not required. This technique is very flexible, and allows the preparation of unique patterns of nanowires which provides opportunities to produce structures with enhanced anisotropic electrical and magnetic properties. An example of this is the unique fabrication of aligned nanowire grids by overlaying layers of nanowires oriented at different angles with respect to each other. The resistivity of printed and cured films was found to be as low as 560 µΩ∙cm. The saturation magnetization was measured to be 30 emu∙g-1, which is comparable to bulk Ni. Magnetic anisotropy was induced with an axis along the direction of the applied magnetic field, giving soft magnetic properties.

A nanoliter resolution implantable micropump for murine inner ear drug delivery.

Advances in protective and restorative biotherapies have created new opportunities to use site-directed, programmable drug delivery systems to treat auditory and vestibular disorders. Successful therapy development that leverages the transgenic, knock-in, and knock-out variants of mouse models of human disease requires advanced microsystems specifically designed to function with nanoliter precision and with system volumes suitable for implantation. Here we present results for a novel biocompatible, implantable, scalable, and wirelessly controlled peristaltic micropump. The micropump configuration included commercially available catheter microtubing (250 µm OD, 125 µm ID) that provided a biocompatible leak-free flow path while avoiding complicated microfluidic interconnects. Peristaltic pumping was achieved by sequentially compressing the microtubing via expansion and contraction of a thermal phase-change material located in three chambers integrated adjacent to the microtubing. Direct-write micro-scale printing technology was used to build the mechanical components of the micropump around the microtubing directly on the back of a printed circuit board assembly (PCBA). The custom PCBA was fabricated using standard commercial processes providing microprocessor control of actuation and Bluetooth wireless communication through an Android application. The results of in vitro characterization indicated that nanoliter resolution control over the desired flow rates of 10-100 nL/min was obtained by changing the actuation frequency. Applying 10× greater than physiological backpressures and ±â€¯3 °C ambient temperature variation did not significantly affect flow rates. Three different micropumps were tested on six mice for in vivo implantation of the catheter microtubing into the round window membrane niche for infusion of a known ototoxic compound (sodium salicylate) at 50 nL/min for 20 min. Real-time shifts in distortion product otoacoustic emission thresholds and amplitudes were measured during the infusion. There were systematic increases in distortion product threshold shifts during the 20-min perfusions; the mean shift was 15 dB for the most basal region. A biocompatibility study was performed to evaluate material suitability for chronic subcutaneous implantation and clinical translational development. The results indicated that the micropump components successfully passed key biocompatibility tests. A micropump prototype was implanted for one month without development of inflammation or infection. Although tested here on the small murine cochlea, this low-cost design and fabrication methodology is scalable for use in larger animals and for clinical applications in children and adults by appropriate scaling of the microtubing diameter and actuator volume.

Inkjet Printed Polyethylene Glycol as a Fugitive Ink for the Fabrication of Flexible Microfluidic Systems.

This paper demonstrates a novel and simple processing technique for the realization of scalable and flexible microfluidic microsystems by inkjet-printing polyethylene-glycol (PEG) as a sacrificial template, followed by embedding in a structural layer (e.g. soft elastomers). The printing technology allows production of an array of PEG droplets simultaneously, reducing cost and manufacturing time. The PEG can be removed through heating above its phase-change temperature after the formation of the structural layer, with hydraulic flow removing the material. The developed technique allows easy modulation of the shape and dimensions of the pattern with the ability to generate complex architectures without using lithography. The method produces robust planar and multilayer microfluidic structures that can be realized on wide range of substrates. Moreover, microfluidics can be realized on other systems (e.g. electrodes and transducers) directly without requiring any bonding or assembling steps, which often limit the materials selection in conventional microfluidic fabrication. Multilayer Polydimethylsiloxane (PDMS) microfluidic channels were created using this technique to demonstrate the capability of the concept to realize flexible microfluidic electronics, drug delivery systems, and lab-on-a-chip devices. By utilizing conductive liquid metals (i.e. EGaIn) as the filling material of the channels, flexible passive resistive components and sensors have been realized.

Reactive Silver Oxalate Ink Composition with Enhanced Curing Conditions for Flexible Substrates.

A solid silver-ligand complex, µ-oxolato-bis(ethylenediaminesilver(I)), was developed for formulating particle-free conductive metal-organic decomposition (MOD) inkjet inks. The complex comprises both a high molar silver content and solubility in inkjet compatible polar solvents. An aqueous ink formulation with 29.5 wt % silver content was developed and inkjet printed onto glass, polyethylene terephthalate, and polyimide substrates. A new hybrid thermal-photonic curing approach resulting in substantially improved electrical properties and substrate adhesion is presented. Silver conductive traces were measured to have bulk resistivity of 4.26 × 10-8 Ω m, which is 2.7 times that of bulk silver. One-pot complex synthesis yielded an easily isolated, and stable, solid product that can be formulated when needed thereby improving shelf life.

Bio-printing cell-laden Matrigel-agarose constructs.

3D printing of biological architectures that mimic the structural and functional features of in vivo tissues is of great interest in tissue engineering and the development of transplantable organ constructs. Printable bio-inks that are compatible with cellular activities play critical roles in the process of 3D bio-printing. Although a variety of hydrogels have been used as bio-inks for 3D bio-printing, they inherit poor mechanical properties and/or the lack of essential protein components that compromise their performance. Here, a hybrid Matrigel-agarose hydrogel system has been demonstrated that possesses both desired rheological properties for bio-printing and biocompatibility for long-term (11 days) cell culture. The agarose component in the hybrid hydrogel system enables the maintenance of 3D-printed structures, whereas Matrigel provides essential microenvironments for cell growth. When human intestinal epithelial HCT116 cells are encapsulated in the printed Matrigel-agarose constructs, high cell viability and proper cell spreading morphology are observed. Given that Matrigel is used extensively for 3D cell culturing, the developed 3D-printable Matrigel-agarose system will open a new way to construct Matrigel-based 3D constructs for cell culture and tissue engineering.

A novel shape memory alloy annuloplasty ring for minimally invasive surgery: design, fabrication, and evaluation.

A novel annuloplasty ring with a shape memory alloy core has been developed to facilitate minimally invasive mitral valve repair. In its activated (austenitic) phase, this prototype ring has comparable mechanical properties to commercial semi-rigid rings. In its pre-activated (martensitic) phase, this ring is flexible enough to be introduced through an 8-mm trocar and easily manipulated with robotic instruments within the confines of a left atrial model. The core is constructed of 0.50 mm diameter NiTi, which is maintained below its martensitic transition temperature (24 °C) during deployment and suturing. After suturing, the ring is heated above its austenitic transition temperature (37 °C, normal human body temperature) enabling the NiTi core to attain its optimal geometry and stiffness characteristics indefinitely. This article summarizes the design, fabrication, and evaluation of this prototype ring. Experimental results suggest that the NiTi core ring could be a viable alternative to flexible bands in robot-assisted minimally invasive mitral valve repair.

Life cycle emissions and cost of producing electricity from coal, natural gas, and wood pellets in Ontario, Canada.

The use of coal is responsible for (1)/(5) of global greenhouse gas (GHG) emissions. Substitution of coal with biomass fuels is one of a limited set of near-term options to significantly reduce these emissions. We investigate, on a life cycle basis, 100% wood pellet firing and cofiring with coal in two coal generating stations (GS) in Ontario, Canada. GHG and criteria air pollutant emissions are compared with current coal and hypothetical natural gas combined cycle (NGCC) facilities. 100% pellet utilization provides the greatest GHG benefit on a kilowatt-hour basis, reducing emissions by 91% and 78% relative to coal and NGCC systems, respectively. Compared to coal, using 100% pellets reduces NO(x) emissions by 40-47% and SO(x) emissions by 76-81%. At $160/metric ton of pellets and $7/GJ natural gas, either cofiring or NGCC provides the most cost-effective GHG mitigation ($70 and $47/metric ton of CO2 equivalent, respectively). The differences in coal price, electricity generation cost, and emissions at the two GS are responsible for the different options being preferred. A sensitivity analysis on fuel costs reveals considerable overlap in results for all options. A lower pellet price ($100/metric ton) results in a mitigation cost of $34/metric ton of CO2 equivalent for 10% cofiring at one of the GS. The study results suggest that biomass utilization in coal GS should be considered for its potential to cost-effectively mitigate GHGs from coal-based electricity in the near term.

A laparoscopic knot-tying device for minimally invasive cardiac surgery.

OBJECTIVE: Intracorporeal suturing and knot tying can complicate, prolong or preclude minimally invasive surgical procedures, reducing their advantages over conventional approaches. An automated knot-tying device has been developed to speed suture fixation during minimally invasive cardiac surgery while retaining the desirable characteristics of conventional hand-tied surgeon's knots: holding strength and visual and haptic feedback. A rotating slotted disc (at the instrument's distal end) automates overhand throws, thereby eliminating the need to manually pass one suture end through a loop in the opposing end. The electronic actuation of this disc produces left or right overhand knots as desired by the operator. METHODS: To evaluate the effectiveness of this technology, seven surgeons with varying laparoscopic experience tied knots within a simulated minimally invasive setting, using both the automated knot-tying tool and conventional laparoscopic tools. Suture types were 2/0 braided and 4/0 monofilament. RESULTS: Mean knot-tying times were 246+/-116 s and 102+/-46 s for conventional and automated methods, respectively, showing an average 56% reduction in time per surgeon (p=0.003, paired t-test). The peak holding strength of each knot (the force required to break the suture or loosen the knot) was measured using tensile-testing equipment. These peak holding strengths were normalised by the ultimate tensile strength of each suture type (57.5 N and 22.1N for 2/0 braided and 4/0 monofilament, respectively). Mean normalised holding strengths for all knots were 68.2% and 71.8% of ultimate tensile strength for conventional and automated methods, respectively (p=0.914, paired t-test). CONCLUSIONS: Experimental data reveal that the automated suturing device has great potential for advancing minimally invasive surgery: it significantly reduced knot-tying times while providing equivalent or greater holding strength than conventionally tied knots.

Evaluation of a shape memory alloy reinforced annuloplasty band for minimally invasive mitral valve repair.

PURPOSE: An in vitro study using explanted porcine hearts was conducted to evaluate a novel annuloplasty band, reinforced with a two-phase, shape memory alloy, designed specifically for minimally invasive mitral valve repair. DESCRIPTION: In its rigid (austenitic) phase, this band provides the same mechanical properties as the commercial semi-rigid bands. In its compliant (martensitic) phase, this band is flexible enough to be introduced through an 8-mm trocar and is easily manipulated within the heart. EVALUATION: In its rigid phase, the prototype band displayed similar mechanical properties to commercially available semi-rigid rings. Dynamic flow testing demonstrated no statistical differences in the reduction of mitral valve regurgitation. In its flexible phase, the band was easily deployed through an 8-mm trocar, robotically manipulated and sutured into place. CONCLUSIONS: Experimental results suggest that the shape memory alloy reinforced band could be a viable alternative to flexible and semi-rigid bands in minimally invasive mitral valve repair.