Photolithographic micropatterning of organic, flexible biomaterials and its applications

A current trend in biodevices has involved a shift from traditional rigid platforms to flexible and stretchable formats. These flexible devices are expected to have a significant impact on future healthcare, disease diagnostics and therapeutics. However, the fabrication of such flexible devices has been limited by the choice of materials. Biomimetic composites of naturally derived and synthetic polymers provide exciting opportunities to develop mechanically flexible, physiologically compliant, and degradable bioelectronic systems. Advantages include the ability to provide conformal contact at non-planar biointerfaces, being able to be degraded at controllable rate, and invoking minimal reactions within the body. These factors present great potential as implantable devices for in-vivo applications, while also addressing concerns with “electronic waste” by being intrinsically degradable. In this work, we present a combination of photo-crosslinkable silk proteins and conductive polymers to precisely fabricate flexible devices and cell culture substrate. A facile and scalable photolithography is applied to fabricate flexible substrates with conductive and non- conductive micropatterns which show tuneable electrical and mechanical properties. We also demonstrate an approach to engineer flexibility in materials through the creation of patterned defects inspired from Kirigami- the Japanese art of paper cutting. Mechanically flexible, free- standing, optically transparent, large-area biomaterial sheets with precisely defined and computationally designed microscale cuts can be formed using a single-step photolithographic process. As composites with conducting polymers, flexible, intrinsically electroactive sheets can be formed. Through this work, the possibility of making next- generation, fully organic, flexible bioelectronics is explored.https://scholarscompass.vcu.edu/gradposters/1099/thumbnail.jp

Effect of substrate stiffness on early human embryonic stem cell differentiation

Background: The pluripotency and self renewing properties of human embryonic stem cells (hESC) make them a valuable tool in the fields of developmental biology, pharmacology and regenerative medicine. Therefore, there exists immense interest in devising strategies for hESC propagation and differentiation. Methods involving simulation of the native stem cell microenvironment, both chemical and physical, have received a lot of attention in recent years. Equally important is evidence that cells can also sense the mechanical properties of their microenvironment. In this study, we test the hypothesis that hESCs accept mechanical cues for differentiation from the substrate by culturing them on flexible polydimethylsiloxane (PDMS) of varying stiffness. Results: PDMS substrates were prepared using available commercial formulations and characterized for stiffness, surface properties and efficiency of cell attachment and proliferation. Across different substrate stiffness, cell numbers, cell attachment and cell surface area were found to be similar. Expression of pluripotency markers decreased with increased time in culture across all PDMS substrates of varying stiffness. Analysis of gene expression of differentiation markers indicates that the differentiation process becomes less stochastic with longer culture times. Conclusions: We evaluated the utility of PDMS substrates for stem cell propagation and substrate mediated differentiation. The stiffness affected gene expression of pluripotent and differentiation markers with results indicating that these substrate systems could potentially be used to direct hESC fate towards early mesodermal lineages. This study suggests that coupled with soluble factors, PDMS substrates could potentially be useful in generating defined populations of differentiated cells.Engineering and Applied Science

Biofunctional Silk Kirigami With Engineered Properties.

The fabrication of multifunctional materials that interface with living environments is a problem of great interest. A variety of structural design concepts have been integrated with functional materials to form biodevices and surfaces for health monitoring. In particular, approaches based on kirigami-inspired cuts can engineer flexibility in materials through the creation of patterned defects. Here, the fabrication of a biodegradable and biofunctional "silk kirigami" material is demonstrated. Mechanically flexible, free-standing, optically transparent, large-area biomaterial sheets with precisely defined and computationally designed microscale cuts can be formed using a single-step photolithographic process. Using modeling techniques, it is shown how cuts can generate remarkable "self-shielding" leading to engineered elastic behavior and deformation. As composites with conducting polymers, flexible, intrinsically electroactive sheets can be formed. Importantly, the silk kirigami sheets are biocompatible, can serve as substrates for cell culture, and be proteolytically resorbed. The unique properties of silk kirigami suggest a host of applications as transient, "green", functional biointerfaces, and flexible bioelectronics

Evolución del diseño de interiores en los grandes Centros Comerciales de Lima Central Sur en las últimas tres décadas

La investigación responde a una problemática que se evidencia a través de una serie de debilidades que repercuten en el diseño de dichos centros comerciales. El conocimiento de nuevas tecnologías para el diseño interior era escaso, no había conocimientos de enchapes, acabados finos, iluminación decorativa y diseño interior en general. Los materiales tampoco eran de gran ayuda, solo se conocían las estructuras comunes, como el cemento y el acero. Tampoco había conocimientos sobre técnicas constructivas

Micropatterned silk-fibroin/eumelanin composite films for bioelectronic applications

There has been growing interest in the use of natural bionanomaterials and nanostructured systems for diverse biomedical applications. Such materials can confer unique functional properties as well as address concerns pertaining to sustainability in production. In this work, we propose the biofabrication of micropatterned silk fibroin/eumelanin composite thin films to be used in electroactive and bioactive applications in bioelectronics and biomedical engineering. Eumelanin is the most common form of melanin, naturally derived from the ink of cuttlefish, having antioxidant and electroactive properties. Another natural biomaterial, the protein silk fibroin, is modified with photoreactive chemical groups, which allows the formation of electroactive eumelanin thin films with different microstructures. The silk fibroin/eumelanin composites are fabricated to obtain thin films as well as electroactive microstructures using UV curing. Here, we report for the first time the preparation, characterization, and physical, electrochemical, and biological properties of these natural silk fibroin/eumelanin composite films. Higher concentrations of eumelanin incorporated into the films exhibit a higher charge storage capacity and good electroactivity even after 100 redox cycles. In addition, the microscale structure and the cellular activity of the fibroin/eumelanin films are assessed for understanding of the biological properties of the composite. The developed micropatterned fibroin/eumelanin films can be applied as natural electroactive substrates for bioapplications (e.g., bioelectronics, sensing, and theranostics) because of their biocompatible properties.The authors acknowledge the FRONTHERA project (Frontiers of technology for theranostics of cancer, metabolic and neurodegenerative diseases) n degrees NORTE-01-0145-FEDER0000232, the European Union Framework Programme for Research and Innovation Horizon 2020 under grant agreement n degrees 668983. FoReCaST (Forefront Research in 3D Disease Cancer Models as in vitro Screening Technologies), and FCT grants POCI-01-0145-FEDER-031590, PD/BD/150546/2019 and PTDC/BTM-ORG/28168/2017. VKY acknowledges support from the National Science Foundation (CBET1704435)

Portable, Disposable, Biomimetic Electrochemical Sensors for Analyte Detection in a Single Drop of Whole Blood

Current diagnostics call for rapid, sensitive, and selective screening of physiologically important biomarkers. Point-of-care (POC) devices for the rapid, reliable, and easy acquisition of bioinformation at, or near the patient, offer opportunities for better healthcare management. Electrochemical biosensors with high sensitivity and ease of miniaturization are advantageous for such applications. We report a photolithographically micropatterned PEDOT:PSS and silk protein-based fully organic 3-electrode sensor (O3ES) for ultralow volume (single drop—10 µL) detection of analytes in whole blood. The O3ES produces reliable electrochemical signals in whole blood from a mouse model with minimal biofouling interference. The O3ES is demonstrated as a portable device for the simultaneous detection of dopamine, ascorbic acid and uric acid using voltammetry techniques. The O3ES displays excellent sensitivity towards each analyte in whole blood, and in the presence of each other. The water-based, ambient processing of the sensors allows the immobilization of enzymes in the organic working electrode. Amperometric detection of uric acid via uricase with high sensitivity in whole blood is demonstrated. Finally, the performance of the O3ES under enzymatic degradation is studied by monitoring sensitivity over an operating lifetime of ~14 days. This work demonstrates the realization of low-cost, disposable POC sensors capable of detecting blood metabolites using ultralow sample volumes

Portable, Disposable, Biomimetic Electrochemical Sensors for Analyte Detection in a Single Drop of Whole Blood

Current diagnostics call for rapid, sensitive, and selective screening of physiologically important biomarkers. Point-of-care (POC) devices for the rapid, reliable, and easy acquisition of bioinformation at, or near the patient, offer opportunities for better healthcare management. Electrochemical biosensors with high sensitivity and ease of miniaturization are advantageous for such applications. We report a photolithographically micropatterned PEDOT:PSS and silk protein-based fully organic 3-electrode sensor (O3ES) for ultralow volume (single drop—10 µL) detection of analytes in whole blood. The O3ES produces reliable electrochemical signals in whole blood from a mouse model with minimal biofouling interference. The O3ES is demonstrated as a portable device for the simultaneous detection of dopamine, ascorbic acid and uric acid using voltammetry techniques. The O3ES displays excellent sensitivity towards each analyte in whole blood, and in the presence of each other. The water-based, ambient processing of the sensors allows the immobilization of enzymes in the organic working electrode. Amperometric detection of uric acid via uricase with high sensitivity in whole blood is demonstrated. Finally, the performance of the O3ES under enzymatic degradation is studied by monitoring sensitivity over an operating lifetime of ~14 days. This work demonstrates the realization of low-cost, disposable POC sensors capable of detecting blood metabolites using ultralow sample volumes

Backbone-branched DNA building blocks for facile angular control in nanostructures

Nanotechnology based on the highly specific pairing of nucleobases in DNA has been used to generate a wide variety of well-defined two- and three-dimensional assemblies, both static and dynamic. However, control over the junction angles to achieve them has been limited. To achieve higher order assemblies, the strands of the DNA duplex are typically made to deviate at junctions with configurations based on crossovers or non-DNA moieties. Such strand crossovers tend to be intrinsically unstructured with the overall structural rigidity determined by the architecture of the nanoassembly, rather than the junction itself. Specific approaches to define nanoassembly junction angles are based either on the cooperative twist- and strain-promoted tuning of DNA persistence length leading to bent DNA rods for fairly large nano-objects, or de novo synthesis of individual junction inserts that are typically non-DNA and based on small organic molecules or metal-coordinating ligand moieties. Here, we describe a general strategy for direct control of junction angles in DNA nanostructures that are completely tunable about the DNA helix. This approach is used to define angular vertices through readily accessible backbone-branched DNAs (bbDNAs). We demonstrate how such bbDNAs can be used as a new building block in DNA nanoconstruction to obtain well-defined nanostructures. Angular control through readily accessible bbDNA building block provides a general and versatile approach for incorporating well-defined junctions in nanoconstructs and expands the toolkit toward achieving strain free, highly size- and shape-tunable DNA based architectures

A Thermal‐Reflow‐Based Low‐Temperature, High‐Pressure Sintering of Lyophilized Silk Fibroin for the Fast Fabrication of Biosubstrates

Solid fibroin is a bulk nonporous material that can be prepared with two methods: a liquid–gel–solid transition from a fibroin solution or a sintering procedure starting from silk powder. Both methods have their own disadvantages: the first requires several weeks and the process is size dependent; the second requires high temperatures. To overcome these limitations, a low‐temperature sintering procedure based on a thermal‐reflow is proposed in this work to produce in fast‐fashion monoliths of solid fibroin. Thermal‐reflow is a well‐known mechanism that takes place when the glass transition temperature of the material is lower than the temperature used to process it. Water plays an important role decreasing the glass transition temperature down to 40 °C. For the first time, a thermal reflow is conducted on lyophilized silk fibroin at 40 °C, associating to the water addition a high‐pressure compression. To optimize the process, a full factorial design of experiment is used. The material is then studied in the crucial phases by digital scanning calorimetry, Fourier‐transform infrared spectroscopy, and scanning electron microscopy. Finally, a mechanical characterization and a preliminary in vitro test are conducted