Fully Integrated 3D Microelectrode Arrays with Polydopamine-Mediated Silicon Dioxide Insulation for Electrophysiological Interrogation of a Novel 3D Human, Neural Microphysiological Construct.

Advances within in vitro biological system complexity have enabled new possibilities for the "Organs-on-a-Chip" field. Microphysiological systems (MPS) as such incorporate sophisticated biological constructs with custom biological sensors. For microelectromechanical systems (MEMS) sensors, the dielectric layer is critical for device performance, where silicon dioxide (SiO2) represents an excellent candidate due to its biocompatibility and wide utility in MEMS devices. Yet, high temperatures traditionally preclude SiO2 from incorporation in polymer-based BioMEMS. Electron-beam deposition of SiO2 may provide a low-temperature, dielectric serving as a nanoporous MPS growth substrate. Herein, we enable improved adherence of nanoporous SiO2 to polycarbonate (PC) and 316L stainless steel (SS) via polydopamine (PDA)-mediated chemistry. The resulting stability of the combinatorial PDA-SiO2 film was interrogated, along with the nature of the intrafilm interactions. A custom polymer-metal three-dimensional (3D) microelectrode array (3D MEA) is then reported utilizing PDA-SiO2 insulation, for definition of novel dorsal root ganglion (DRG)/nociceptor and dorsal horn (DH) 3D neural constructs in excess of 6 months for the first time. Spontaneous/evoked compound action potentials (CAPs) are successfully reported. Finally, inhibitory drugs treatments showcase pharmacological responsiveness of the reported multipart biological activity. These results represent the initiation of a novel 3D MEA-integrated, 3D neural MPS for the long-term electrophysiological study.

Biphasic burst and sustained transdermal delivery in vivo using an AI-optimized 3D-printed MN patch.

The objective of the present study was to fabricate microneedles for delivering lipophilic active ingredients (APIs) using digital light processing (DLP) printing technology and quality by design (QbD) supplemented by artificial intelligence (AI) algorithms. In the present study, dissolvable microneedle (MN) patches using ibuprofen (IBU) as a model drug were successfully fabricated with DLP printing technology at âˆ¼ 750 µm height, ∼250 µm base diameter, and tip with radius of curvature (RoC) of âˆ¼ 15 µm. MN patches were comprised of IBU, photoinitiator, Lithium phenyl (2,4,6-trimethylbenzoyl) phosphinate (LAP), polyethylene glycol dimethacrylate (PEGDAMA)550 and distilled water and were developed using the QbD optimization approach. Optimization of print fidelity and needle morphology were achieved using AI implementing a semi-supervised machine learning approach. Mechanical strength tests demonstrated that IBU MNs formed pores both on Parafilm M® and human cadaver skin. IBU-MNs consisting of 0.23 %w/v and 0.49 %w/v LAP with 10 %w/v water showed âˆ¼ 2 mg/cm2 sustained drug permeation at 72 h in skin permeation experiments with flux of âˆ¼ 40 µg/cm2/h. Pharmacokinetic studies in rats displayed biphasic rapid first-order absorption with sustained zero-order input of Ko = 150ug/hr, AUC0-48h = 62812.02 ± 11128.39 ng/ml*h, Tmax = 2.66 ± 1.12 h, and Cmax = 3717.43 ± 782.25 ng/ml (using 0.23 %w/v LAP IBU MN patch). An in vitro in vivo relation (IVIVR) was conducted identifying a polynomial relationship between patch release and fraction absorbed in vivo. This study demonstrates fabrication of dissolvable DLP-printed microneedle patches for lipophilic API delivery with biphasic rapid first-order and sustained zero-order release.

Rapid Makerspace Microfabrication and Characterization of 3D Microelectrode Arrays (3D MEAs) for Organ-on-a-Chip Models.

Integrated sensors in "on-a-chip" in vitro cellular models are a necessity for granularity in data collection required for advanced biosensors. As these models become more complex, the requirement for the integration of electrogenic cells is apparent. Interrogation of such cells, whether alone or within a connected cellular framework, are best achieved with microelectrodes, in the form of a microelectrode array (MEA). Makerspace microfabrication has thus far enabled novel and accessible approaches to meet these demands. Here, resin-based 3D printing, selective multimodal laser micromachining, and simple insulation strategies, define an approach to highly customizable and "on-demand" in vitro 3D MEA-based biosensor platforms. The scalability of this approach is aided by a novel makerspace microfabrication assisted technique denoted using the term Hypo-Rig. The MEA utilizes custom-defined metal microfabricated microelectrodes transitioned from planar (2D) to 3D using the Hypo-Rig. To simulate this transition process, COMSOL modeling is utilized to estimate transitionary forces and angles (with respect to normal). Practically, the Hypo-Rig demonstrated a force of ~40N, as well as a consistent 70° average angular transitionary performance which matched well with the COMSOL model. To illustrate the scalability potential, 3 × 3, 6 × 6, and 8 × 8 versions of the device were fabricated and characterized. The 3D MEAs, demonstrated impedance and phase measurements in the biologically relevant 1 kHz range of 45.4 kΩ, and -34.6° respectively, for polystyrene insulated, ~70µm sized microelectrodes.

Fabrication and Characterization of 3D Printed, 3D Microelectrode Arrays for Interfacing with a Peripheral Nerve-on-a-Chip.

We present a nontraditional fabrication technique for the realization of three-dimensional (3D) microelectrode arrays (MEAs) capable of interfacing with 3D cellular networks in vitro. The technology uses cost-effective makerspace microfabrication techniques to fabricate the 3D MEAs with 3D printed base structures with the metallization of the microtowers and conductive traces being performed by stencil mask evaporation techniques. A biocompatible lamination layer insulates the traces for realization of 3D microtower MEAs (250 µm base diameter, 400 µm height). The process has additionally been extended to realize smaller electrodes (30 µm × 30 µm) at a height of 400 µm atop the 3D microtower using laser micromachining of an additional silicon dioxide (SiO2) insulation layer. A 3D microengineered, nerve-on-a-chip in vitro model for recording and stimulating electrical activity of dorsal root ganglion (DRG) cells has further been integrated with the 3D MEA. We have characterized the 3D electrodes for electrical, chemical, electrochemical, biological, and chip hydration stability performance metrics. A decrease in impedance from 1.8 kΩ to 670 Ω for the microtower electrodes and 55 to 39 kΩ for the 30 µm × 30 µm microelectrodes can be observed for an electrophysiologically relevant frequency of 1 kHz upon platinum electroless plating. Biocompatibility assays on the components of the system resulted in a large range (∼3%-70% live cells), depending on the components. Fourier-transform infrared (FTIR) spectra of the resin material start to reveal possible compositional clues for the resin, and the hydration stability is demonstrated in in-vitro-like conditions for 30 days. The fabricated 3D MEAs are rapidly produced with minimal usage of a cleanroom and are fully functional for electrical interrogation of the 3D organ-on-a-chip models for high-throughput of pharmaceutical screening and toxicity testing of compounds in vitro.

Makerspace microfabrication of a stainless steel 3D microneedle electrode array (3D MEA) on a glass substrate for simultaneous optical and electrical probing of electrogenic cells.

Microfabrication and assembly of a Three-Dimensional Microneedle Electrode Array (3D MEA) based on a glass-stainless steel platform is demonstrated involving the utilization of non-traditional "Makerspace Microfabrication" techniques featuring cost-effective, rapid fabrication and an assorted biocompatible material palette. The stainless steel microneedle electrode array was realized by planar laser micromachining and out-of-plane transitioning to have a 3D configuration with perpendicular transition angles. The 3D MEA chip is bonded onto a glass die with metal traces routed to the periphery of the chip for electrical interfacing. Confined precision drop casting (CPDC) of PDMS is used to define an insulation layer and realize the 3D microelectrodes. The use of glass as a substrate offers optical clarity allowing for simultaneous optical and electrical probing of electrogenic cells. Additionally, an interconnect using 3D printing and conductive ink casting has been developed which allows metal traces on the glass chip to be transitioned to the bottomside of the device for interfacing with commercial data acquisition/analysis equipment. The 3D MEAs demonstrate an average impedance/phase of ∼13.3 kΩ/-12.1° at 1 kHz respectively, and an average 4.2 µV noise. Lastly, electrophysiological activity from an immortal cardiomyocyte cell line was recorded using the 3D MEA demonstrating end to end device development.

Capabilities and limitations of 3D printed microserpentines and integrated 3D electrodes for stretchable and conformable biosensor applications.

We explore the capabilities and limitations of 3D printed microserpentines (µserpentines) and utilize these structures to develop dynamic 3D microelectrodes for potential applications in in vitro, wearable, and implantable microelectrode arrays (MEAs). The device incorporates optimized 3D printed µserpentine designs with out-of-plane microelectrode structures, integrated on to a flexible Kapton® package with micromolded PDMS insulation. The flexibility of the optimized, printed µserpentine design was calculated through effective stiffness and effective strain equations, so as to allow for analysis of various designs for enhanced flexibility. The optimized, down selected µserpentine design was further sputter coated with 7-70 nm-thick gold and the performance of these coatings was studied for maintenance of conductivity during uniaxial strain application. Bending/conforming analysis of the final devices (3D MEAs with a Kapton® package and PDMS insulation) were performed to qualitatively assess the robustness of the finished device toward dynamic MEA applications. 3D microelectrode impedance measurements varied from 4.2 to 5.2 kΩ during the bending process demonstrating a small change and an example application with artificial agarose skin composite model to assess feasibility for basic transdermal electrical recording was further demonstrated.

SeedEZ™ Interdigitated Electrodes and Multifunctional Layered Biosensor Composites (MLBCs): A Paradigm Shift in the Development of In Vitro BioMicrosystems.

We have developed a new technology for the realization of composite biosensor systems, capable of measuring electrical and electrophysiological signals from electrogenic cells, using SeedEZ™ 3D cell culture-scaffold material. This represents a paradigm-shift for BioMEMS processing; 'Biology-Microfabrication' versus the standard 'Microfabrication-Biology' approach. An Interdigitated Electrode (IDE) developed on the 3D cell-scaffold was used to successfully monitor acute cardiomyocyte growth and controlled population decline. We have further characterized processability of the 3D scaffold, demonstrated long-term biocompatibility of the scaffold with various cell lines and developed a multifunctional layered biosensor composites (MLBCs) using SeedEZ™ and other biocompatible substrates for future multilayer sensor integration.

Precision Vascular Delivery of Agrochemicals with Micromilled Microneedles (µMMNs).

We demonstrate use of makerspace techniques involving subtractive microtechnologies to fabricate micromilled microneedles (µMMNs) of stainless steel (SS) for precise delivery of agrochemicals into vascular bundles of plant tissue. Precision delivery is of immense importance for systemic pathogen control in specific areas of plant tissue. Optimization of the micromilling allows for selective removal of SS at the microscale and the microfabrication of a 5 × 5 array of µMMNs having both base width and height of 500 µm to enable precise puncture into the stem of citrus saplings. Atomic Absorption Spectroscopy reveals up to 7.5× increase in the uptake of a therapeutic cargo while Scanning Electron Microscopy reveals that specific sites of the vascular bundle; either xylem or the phloem can be uniquely targeted with customized µMMNs. Such rapid and cost-effective customization with intricate designs along with scalability is enabled by makerspace microfabrication. Additionally, a 19 × 20 array of micromilled mesoneedles has been fabricated and affixed to a paint roller as an applicator system for real-world field testing outside the laboratory. Initial results indicate reliable behavior of the applicator system and the technique can be applied to the systemic delivery of agrochemicals while conserving the loss of the agrochemical with increased application efficiency.

Optimization of makerspace microfabrication techniques and materials for the realization of planar, 3D printed microelectrode arrays in under four days.

Conventional two-dimensional microelectrode arrays (2D MEAs) in the market involve long manufacturing timeframes, have cleanroom requirements, and need to be assembled from multiple parts to obtain the final packaged device. For MEAs to be "used and tossed", manufacturing has to be moved from the cleanroom to makerspaces. In order to enable makerspace fabricated MEAs comparable to conventional MEAs, the microfabrication processes must be optimized to have similar electrical properties along with biocompatibility and number of recording sites. This work presents a makerspace microfabricated 2D MEA having electrode densities up to a commercially popular 8 × 8 array, all fabricated under four days. Additive manufacturing-based realization of the MEA devices provides immense flexibility in terms of meeting distinct design requirements. A unique non-planar MEA having meso-scale electrodes on the top side of a chip transitioning to traces onto the bottom side through electrical vias is presented in this work. This allows for (a) monolithic integration of a culture well for devices having up to a 6 × 6 MEA array, (b) selective electroplating of the meso-scale electrodes (500 µm diameter) defined by silver ink casting followed by pulsed electroplating of gold or platinum without any masking procedure, (c) casting of a uniform and planar insulation layer via a novel process of confined precision spin coating (CPSC) of SU-8 which acts as a biocompatible insulation atop the meso-scale electrodes; and (d) selective laser micromachining to define the 50 µm × 50 µm microelectrodes. For an 8 × 8 array, the culture well and MEA chip framework are 3D printed as two separate parts and sealed together with a biocompatible epoxy as in commercially available MEAs. The fabricated MEAs have an average 1 kHz impedance of 36.8 kΩ/16 kΩ with a double layer capacitance of 400 nF cm-2/520 nF cm-2 for nano-porous platinum/nano-gold which is comparable to the state-of-art commercially available 2D MEAs. Additionally, it was found out that our 3D printing-based process compares very favorably with traditional glass MEAs in terms of design to device while representing a dramatic reduction in cost, timeline for fabrication, reduction in the number of steps and the need for sophisticated microfabrication and packaging equipment.

3D Printing, Ink Casting and Micromachined Lamination (3D PICLµM): A Makerspace Approach to the Fabrication of Biological Microdevices.

We present a novel benchtop-based microfabrication technology: 3D printing, ink casting, micromachined lamination (3D PICLµM) for rapid prototyping of lab-on-a-chip (LOC) and biological devices. The technology uses cost-effective, makerspace-type microfabrication processes, all of which are ideally suited for low resource settings, and utilizing a combination of these processes, we have demonstrated the following devices: (i) 2D microelectrode array (MEA) targeted at in vitro neural and cardiac electrophysiology, (ii) microneedle array targeted at drug delivery through a transdermal route and (iii) multi-layer microfluidic chip targeted at multiplexed assays for in vitro applications. The 3D printing process has been optimized for printing angle, temperature of the curing process and solvent polishing to address various biofunctional considerations of the three demonstrated devices. We have depicted that the 3D PICLµM process has the capability to fabricate 30 µm sized MEAs (average 1 kHz impedance of 140 kΩ with a double layer capacitance of 3 µF), robust and reliable microneedles having 30 µm radius of curvature and ~40 N mechanical fracture strength and microfluidic devices having 150 µm wide channels and 400 µm fluidic vias capable of fluid mixing and transmitted light microparticle visualization. We believe our 3D PICLµM is ideally suited for applications in areas such as electrophysiology, drug delivery, disease in a dish, organ on a chip, environmental monitoring, agricultural therapeutic delivery and genomic testing.

Rapid Nanofabrication of Nanostructured Interdigitated Electrodes (nIDEs) for Long-Term In Vitro Analysis of Human Induced Pluripotent Stem Cell Differentiated Cardiomyocytes.

Adverse cardiac events are a major cause of late-stage drug development withdrawals. Improved in vitro systems for predicting cardiotoxicity are of great interest to prevent these events and to reduce the expenses involved in the introduction of cardiac drugs into the marketplace. Interdigitated electrodes (IDEs) affixed with a culture well provide a simple, suitable solution for in vitro analysis of cells because of their high sensitivity, ease of fabrication, and label-free, nondestructive analysis. Culturing human pluripotent stem cell differentiated cardiomyocytes onto these IDEs allows for the use of the IDE⁻cell combination in predictive toxicity assays. IDEs with smaller interdigitated distances allow for greater sensitivity, but typically require cleanroom fabrication. In this communication, we report the definition of a simple IDE geometry on a printed nanostructured substrate, demonstrate a Cellular Index (CI) increase from 0 to 7.7 for human cardiomyocytes, and a decrease in CI from 2.3 to 1 with increased concentration of the model drug, norepinephrine. The nanostructuring results in an increased sensitivity of our 1 mm pitch IDEs when compared to traditionally fabricated IDEs with a pitch of 10 µm (100 times larger electrode gap). The entire nanostructured IDE (nIDE) is fabricated and assembled in a rapid nanofabrication environment, thus allowing for iterative design changes and robust fabrication of devices.

c-Si/n-ZnO-based flexible solar cells with silica nanoparticles as a light trapping metamaterial.

Herein, we report the fabrication of flexible solar cells based on a crystalline p-Si/n-ZnO heterostructure for the first time. An enhancement of ∼52% in the base efficiency was achieved by the application of spherical SiO2 nanoparticles as light trapping structures on the top. The use of ZnO not only offers a facile route of synthesis, but also provides an additional advantage of large band bending, leading to notable open circuit voltage and formation of an intermediate ultra-thin barrier layer of Zn2SiO4 for minimized carrier recombination. The spherical silica nanoparticles act as nanoresonators, causing absorption hot-spots in the thin silicon absorber, along with the capability of providing wide-angle light-collection. Simulation showed, for the higher angle of incidence, that the silica nanoparticles have the ability to bend light on the same side of the normal to the incident wave-front, thereby acting as a negative index metamaterial (NIM). The flexibility and cost-effectiveness of this device can make it important for the next-generation photovoltaics and roll-to-roll electronics.

Black silicon solar cell: analysis optimization and evolution towards a thinner and flexible future.

Analysis and optimization of silicon nano-structured geometry (black silicon) for photovoltaic applications has been reported. It is seen that a unique class of geometry: micro-nanostructure has the potential to find a balance between the conflicting interests of reduced reflection for wide angles of incidence, reduced surface area enhancement due to the nano-structuring of the substrate and reduced material wastage due to the etching of the silicon substrate to realize the geometry itself. It is established that even optimally designed micro-nanostructures would not be useful for conventional wafer based approaches. The work presents computational studies on how such micro-nanostructures are more potent for future ultra-thin monocrystalline silicon absorbers. For such ultra-thin absorbers, the optimally designed micro-nanostructures provide additional advantages of advanced light management capabilities as it behaves as a lossy 2D photonic crystal making the physically thin absorber optically thick along with the ability to collect photo-generated carriers orthogonal to the direction of light (radial junction) for unified photon-electron harvesting. Most significantly, the work answers the key question on how thin the monocrystalline solar absorber should be so that optimum micro-nanostructure would be able to harness the incident photons ensuring proper collection so as to reach the well-known Shockley-Queisser limit of solar cells. Flexible ultra-thin monocrystalline silicon solar cells have been fabricated using nanosphere lithography and MacEtch technique along with a synergistic association of crystalline and amorphous silicon technologies to demonstrate its physical and technological flexibilities. The outcomes are relevant so that nanotechnology may be seamlessly integrated into the technology roadmap of monocrystalline silicon solar cells as the silicon thickness should be significantly reduced without compromising the efficiency within the next decade.