Pressure Stimuli and Spatiotemporal Illusions on the Forearm.

To design complex wearable haptic interfaces using pressure, we have to explore how we can use pressure stimuli to their full potential. Haptic illusions, such as apparent motion and apparent location, can be a part of this. If these illusions can be evoked with pressure, haptic patterns can increase in complexity without increasing the number of actuators or combining different types of actuators. We did two psychophysical experiments with pressure stimuli on the forearm using a pneumatic sleeve with multiple, individually controlled McKibben actuators. In Experiment 1, we found that spatial integration of two simultaneously presented stimuli occurred for distances up to 61 mm. In Experiment 2, we found that apparent motion can be elicited with distinct pressure stimuli over a range of temporal parameters. These results clearly show spatio-temporal integration in the somatosensory system for pressure stimuli. We discuss these findings in relation to effects found for vibration and the mechanoreceptors in the glabrous skin.

Electrochemical control of bone microstructure on electroactive surfaces for modulation of stem cells and bone tissue engineering.

Controlling stem cell behavior at the material interface is crucial for the development of novel technologies in stem cell biology and regenerative medicine. The composition and presentation of bio-factors on a surface strongly influence the activity of stem cells. Herein, we designed an electroactive surface that mimics the initial process of trabecular bone formation, by immobilizing chondrocyte-derived plasma membrane nanofragments (PMNFs) on its surface for rapid mineralization within 2 days. Moreover, the electroactive surface was based on the conducting polymer polypyrrole (PPy), which enabled dynamic control of the presentation of PMNFs on the surface via electrochemical redox switching, further resulting in the formation of bone minerals with different morphologies. Furthermore, bone minerals with contrasting surface morphologies had differential effects on the differentiation of human bone marrow-derived stem cells (hBMSCs) cultured on the surface. Together, this electroactive surface showed multifunctional characteristics, not only allowing dynamic control of PMNF presentation but also promoting the formation of bone minerals with different morphologies within 2 days. This electroactive substrate could be valuable for more precise control of stem cell growth and differentiation, and further development of more suitable microenvironments containing bone apatite for housing a bone marrow stem cell niche, such as biochips/bone-on-chips.

Plant-Like Tropisms in Artificial Muscles.

Helical plants have the ability of tropisms to respond to natural stimuli, and biomimicry of such helical shapes into artificial muscles has been vastly popular. However, the shape-mimicked actuators only respond to artificially provided stimulus, they are not adaptive to variable natural conditions, thus being unsuitable for real-life applications where on-demand, autonomous operations are required. Novel artificial muscles made of hierarchically patterned helically wound yarns that are self-adaptive to environmental humidity and temperature changes are demonstrated here. Unlike shape-mimicked artificial muscles, a unique microstructural biomimicking approach is adopted, where the muscle yarns can effectively replicate the hydrotropism and thermotropism of helical plants to their microfibril level using plant-like microstructural memories. Large strokes, with rapid movement, are obtained when the individual microfilament of yarn is inlaid with hydrogel and further twisted into a coil-shaped hierarchical structure. The developed artificial muscle provides an average actuation speed of ≈5.2% s-1 at expansion and ≈3.1% s-1 at contraction cycles, being the fastest amongst previously demonstrated actuators of similar type. It is demonstrated that these muscle yarns can autonomously close a window in wet climates. The building block yarns are washable without any material degradation, making them suitable for smart, reusable textile and soft robotic devices.

Biohybrid Variable-Stiffness Soft Actuators that Self-Create Bone.

Inspired by the dynamic process of initial bone development, in which a soft tissue turns into a solid load-bearing structure, the fabrication, optimization, and characterization of bioinduced variable-stiffness actuators that can morph in various shapes and change their properties from soft to rigid are hereby presented. Bilayer devices are prepared by combining the electromechanically active properties of polypyrrole with the compliant behavior of alginate gels that are uniquely functionalized with cell-derived plasma membrane nanofragments (PMNFs), previously shown to mineralize within 2 days, which promotes the mineralization in the gel layer to achieve the soft to stiff change by growing their own bone. The mineralized actuator shows an evident frozen state compared to the movement before mineralization. Next, patterned devices show programmed directional and fixated morphing. These variable-stiffness devices can wrap around and, after the PMNF-induced mineralization in and on the gel layer, adhere and integrate onto bone tissue. The developed biohybrid variable-stiffness actuators can be used in soft (micro-)robotics and as potential tools for bone repair or bone tissue engineering.

3D Printing Microactuators for Soft Microrobots.

Current additive manufacturing, including three-dimensional (3D) and so-called four-dimensional printing, of soft robotic devices is limited to millimeter sizes. In this study, we present additive manufacturing of soft microactuators and microrobots to fabricate even smaller structures in the micrometer domain. Using a custom-built extrusion 3D printer, microactuators are scaled down to a size of 300 × 1000 µm2, with minimum thickness of 20 µm. Microactuators combined with printed body and electroactive polymers to drive the actuators are fabricated from computer-aided design model of the device structure. To demonstrate the ease and versatility of 3D printing process, microactuators with varying lengths ranging from 1000 to 5000 µm are fabricated and operated. Likewise, microrobotic devices consisting of a rigid body and individually controlled free-moving arms or legs are 3D printed to explore the microfabrication of soft grippers, manipulators, or microrobots through simple additive manufacturing technique.

Novel fabrication of soft microactuators with morphological computing using soft lithography.

A simple and cost-effective method for the patterning and fabrication of soft polymer microactuators integrated with morphological computation is presented. The microactuators combine conducting polymers to provide the actuation, with spatially designed structures for a morphologically controlled, user-defined actuation. Soft lithography is employed to pattern and fabricate polydimethylsiloxane layers with geometrical pattern, for use as a construction element in the microactuators. These microactuators could obtain multiple bending motions from a single fabrication process depending on the morphological pattern defined in the final step. Instead of fabricating via conventional photolithography route, which involves multiple steps with different chromium photomasks, this new method uses only one single design template to produce geometrically patterned layers, which are then specifically cut to obtain multiple device designs. The desired design of the actuator is decided in the final step of fabrication. The resulting microactuators generate motions such as a spiral, screw, and tube, using a single design template.

Artificial Muscles Powered by Glucose.

Untethered actuation is important for robotic devices to achieve autonomous motion, which is typically enabled by using batteries. Using enzymes to provide the required electrical charge is particularly interesting as it will enable direct harvesting of fuel components from a surrounding fluid. Here, a soft artificial muscle is presented, which uses the biofuel glucose in the presence of oxygen. Glucose oxidase and laccase enzymes integrated in the actuator catalytically convert glucose and oxygen into electrical power that in turn is converted into movement by the electroactive polymer polypyrrole causing the actuator to bend. The integrated bioelectrode pair shows a maximum open-circuit voltage of 0.70 ± 0.04 V at room temperature and a maximum power density of 0.27 µW cm-2 at 0.50 V, sufficient to drive an external polypyrrole-based trilayer artificial muscle. Next, the enzymes are fully integrated into the artificial muscle, resulting in an autonomously powered actuator that can bend reversibly in both directions driven by glucose and O2 only. This autonomously powered artificial muscle can be of great interest for soft (micro-)robotics and implantable or ingestible medical devices manoeuvring throughout the body, for devices in regenerative medicine, wearables, and environmental monitoring devices operating autonomously in aqueous environments.

Conjugated Polymer Actuators and Devices: Progress and Opportunities.

Conjugated polymers (CPs), as exemplified by polypyrrole, are intrinsically conducting polymers with potential for development as soft actuators or "artificial muscles" for numerous applications. Significant progress has been made in the understanding of these materials and the actuation mechanisms, aided by the development of physical and electrochemical models. Current research is focused on developing applications utilizing the advantages that CP actuators have (e.g., low driving potential and easy to miniaturize) over other actuating materials and on developing ways of overcoming their inherent limitations. CP actuators are available as films, filaments/yarns, and textiles, operating in liquids as well as in air, ready for use by engineers. Here, the milestones made in understanding these unique materials and their development as actuators are highlighted. The primary focus is on the recent progress, developments, applications, and future opportunities for improvement and exploitation of these materials, which possess a wealth of multifunctional properties.

Highly Conductive, Photolithographically Patternable Ionogels for Flexible and Stretchable Electrochemical Devices.

An ionic conducting membrane is an essential part in various electrochemical devices including ionic actuators. To miniaturize these devices, micropatterns of ionic conducting membrane are desired. Here, we present a novel type of ionogel that can be patterned using standard photolithography and soft imprinting lithography. The ionogel is prepared in situ by UV-initiated free-radical polymerization of thiol acrylate precursors in the presence of ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide. The resultant ionogel is very flexible with a low Young's modulus (as low as 0.23 MPa) and shows a very high ionic conductivity (up to 2.4 × 10-3 S/cm with 75 wt % ionic liquid incorporated) and has a reactive surface due to the excess thiol groups. Micropatterns of ionogel are obtained by using the thiol acrylate ionogel solution as an ionic conducting photoresist with standard photolithography. Water, a solvent immiscible with ionic liquid, is used as the photoresist developer to avoid complete removal of ionic liquid from thin micropatterns of the ionogel. By taking advantage of the reactive surface of ionogels and the photopatternability, ionogels with complex three-dimensional microstructure are developed. The surface of the ionogels can also be easily patterned using UV-assisted soft imprinting lithography. This new type of ionogels may open up for building high-performance flexible electrochemical microdevices.

Type I Collagen-Derived Injectable Conductive Hydrogel Scaffolds as Glucose Sensors.

The advent of home blood glucose monitoring revolutionized diabetes management, and the recent introduction of both wearable devices and closed-loop continuous systems has enormously impacted the lives of people with diabetes. We describe the first fully injectable soft electrochemical glucose sensor for in situ monitoring. Collagen, the main component of a native extracellular matrix in humans and animals, was used to fabricate an in situ gellable self-supporting electroconductive hydrogel that can be injected onto an electrode surface or into porcine meat to detect glucose amperometrically. The study provides a proof-of-principle of an injectable electrochemical sensor suitable for monitoring tissue glucose levels that may, with further development, prove clinically useful in the future.

Patterning Highly Conducting Conjugated Polymer Electrodes for Soft and Flexible Microelectrochemical Devices.

There is a need for soft actuators in various biomedical applications to manipulate delicate objects such as cells and tissues. Soft actuators are able to adapt to any shape and limit the stress applied to delicate objects. Conjugated polymer (CP) actuators, especially in the so-called trilayer configuration, are interesting candidates for driving such micromanipulators. However, challenges involved in patterning the electrodes in a trilayer with individual contact have prevented further development of soft micromanipulators based on CP actuators. To allow such patterning, two printing-based patterning techniques have been developed. First, an oxidant layer is printed using either syringe-based printing or microcontact printing, followed by vapor-phase polymerization of the CP. Submillimeter patterns with electronic conductivities of 800 S·cm-1 are obtained. Next, laser ablation is used to cleanly cut the final device structures including the printed patterns, resulting in fingers with individually controllable digits and miniaturized hands. The methods presented in this paper will enable integration of patterned electrically active CP layers in many types of complex three-dimensional structures.

The role of ATP signalling in response to mechanical stimulation studied in T24 cells using new microphysiological tools.

The capacity to store urine and initiate voiding is a valued characteristic of the human urinary bladder. To maintain this feature, it is necessary that the bladder can sense when it is full and when it is time to void. The bladder has a specialized epithelium called urothelium that is believed to be important for its sensory function. It has been suggested that autocrine ATP signalling contributes to this sensory function of the urothelium. There is well-established evidence that ATP is released via vesicular exocytosis as well as by pannexin hemichannels upon mechanical stimulation. However, there are still many details that need elucidation and therefore there is a need for the development of new tools to further explore this fascinating field. In this work, we use new microphysiological systems to study mechanostimulation at a cellular level: a mechanostimulation microchip and a silicone-based cell stretcher. Using these tools, we show that ATP is released upon cell stretching and that extracellular ATP contributes to a major part of Ca2+ signalling induced by stretching in T24 cells. These results contribute to the increasing body of evidence for ATP signalling as an important component for the sensory function of urothelial cells. This encourages the development of drugs targeting P2 receptors to relieve suffering from overactive bladder disorder and incontinence.

Switchable presentation of cytokines on electroactive polypyrrole surfaces for hematopoietic stem and progenitor cells.

Hematopoietic stem cells are used in transplantations for patients with hematologic malignancies. Scarce sources require efficient strategies of expansion, including polymeric biomaterials mimicking architectures of bone marrow tissue. Tissue microenvironment and mode of cytokine presentation strongly influence cell fate. Although several cytokines with different functions as soluble or membrane-bound mediators have already been identified, their precise roles have not yet been clarified. A need exists for in vitro systems that mimic the in vivo situation to enable such studies. One way is to establish surfaces mimicking physiological presentation using protein-immobilization onto polymer films. However these films merely provide a static presentation of the immobilized proteins. It would be advantageous to also dynamically change protein presentation and functionality to better reflect the in vivo conditions. The electroactive polymer polypyrrole shows excellent biocompatibility and electrochemically alters its surface properties, becoming an interesting choice for such setups. Here, we present an in vitro system for switchable presentation of membrane-bound cytokines. We use interleukin IL-3, known to affect hematopoiesis, and show that when immobilized on polypyrrole films, IL-3 is bioavailable for the bone marrow-derived FDC-P1 progenitor cell line. Moreover, IL-3 presentation can be successfully altered by changing the redox state of the film, in turn influencing FDC-P1 cell viability. This novel in vitro system provides a valuable tool for stimuli-responsive switchable protein presentation allowing the dissection of relevant mediators in stem and progenitor cell behavior.

Knitting and weaving artificial muscles.

A need exists for artificial muscles that are silent, soft, and compliant, with performance characteristics similar to those of skeletal muscle, enabling natural interaction of assistive devices with humans. By combining one of humankind's oldest technologies, textile processing, with electroactive polymers, we demonstrate here the feasibility of wearable, soft artificial muscles made by weaving and knitting, with tunable force and strain. These textile actuators were produced from cellulose yarns assembled into fabrics and coated with conducting polymers using a metal-free deposition. To increase the output force, we assembled yarns in parallel by weaving. The force scaled linearly with the number of yarns in the woven fabric. To amplify the strain, we knitted a stretchable fabric, exhibiting a 53-fold increase in strain. In addition, the textile construction added mechanical stability to the actuators. Textile processing permits scalable and rational production of wearable artificial muscles, and enables novel ways to design assistive devices.

Electrochemical bacterial detection using poly(3-aminophenylboronic acid)-based imprinted polymer.

Biosensors can deliver the rapid bacterial detection that is needed in many fields including food safety, clinical diagnostics, biosafety and biosecurity. Whole-cell imprinted polymers have the potential to be applied as recognition elements in biosensors for selective bacterial detection. In this paper, we report on the use of 3-aminophenylboronic acid (3-APBA) for the electrochemical fabrication of a cell-imprinted polymer (CIP). The use of a monomer bearing a boronic acid group, with its ability to specifically interact with cis-diol, allowed the formation of a polymeric network presenting both morphological and chemical recognition abilities. A particularly beneficial feature of the proposed approach is the reversibility of the cis-diol-boronic group complex, which facilitates easy release of the captured bacterial cells and subsequent regeneration of the CIP. Staphylococcus epidermidis was used as the model target bacteria for the CIP and electrochemical impedance spectroscopy (EIS) was explored for the label-free detection of the target bacteria. The modified electrodes showed a linear response over the range of 103-107cfu/mL. A selectivity study also showed that the CIP could discriminate its target from non-target bacteria having similar shape. The CIPs had high affinity and specificity for bacterial detection and provided a switchable interface for easy removal of bacterial cell.

Doping Polypyrrole Films with 4-N-Pentylphenylboronic Acid to Enhance Affinity towards Bacteria and Dopamine.

Here we demonstrate the use of a functional dopant as a fast and simple way to tune the chemical affinity and selectivity of polypyrrole films. More specifically, a boronic-functionalised dopant, 4-N-Pentylphenylboronic Acid (PBA), was used to provide to polypyrrole films with enhanced affinity towards diols. In order to prove the proposed concept, two model systems were explored: (i) the capture and the electrochemical detection of dopamine and (ii) the adhesion of bacteria onto surfaces. The chemisensor, based on overoxidised polypyrrole boronic doped film, was shown to have the ability to capture and retain dopamine, thus improving its detection; furthermore the chemisensor showed better sensitivity in comparison with overoxidised perchlorate doped films. The adhesion of bacteria, Deinococcus proteolyticus, Escherichia coli, Streptococcus pneumoniae and Klebsiella pneumoniae, onto the boric doped polypyrrole film was also tested. The presence of the boronic group in the polypyrrole film was shown to favour the adhesion of sugar-rich bacterial cells when compared with a control film (Dodecyl benzenesulfonate (DBS) doped film) with similar morphological and physical properties. The presented single step synthesis approach is simple and fast, does not require the development and synthesis of functional monomers, and can be easily expanded to the electrochemical, and possibly chemical, fabrication of novel functional surfaces and interfaces with inherent pre-defined sensing and chemical properties.

Plasmonic Metasurfaces with Conjugated Polymers for Flexible Electronic Paper in Color.

A flexible electronic paper in full color is realized by plasmonic metasurfaces with conjugated polymers. An ultrathin large-area electrochromic material is presented which provides high polarization-independent reflection, strong contrast, fast response time, and long-term stability. This technology opens up for new electronic readers and posters with ultralow power consumption.

Direct Mechanical Stimulation of Stem Cells: A Beating Electromechanically Active Scaffold for Cardiac Tissue Engineering.

The combination of stem cell therapy with a supportive scaffold is a promising approach to improving cardiac tissue engineering. Stem cell therapy can be used to repair nonfunctioning heart tissue and achieve myocardial regeneration, and scaffold materials can be utilized in order to successfully deliver and support stem cells in vivo. Current research describes passive scaffold materials; here an electroactive scaffold that provides electrical, mechanical, and topographical cues to induced human pluripotent stem cells (iPS) is presented. The poly(lactic-co-glycolic acid) fiber scaffold coated with conductive polymer polypyrrole (PPy) is capable of delivering direct electrical and mechanical stimulation to the iPS. The electroactive scaffolds demonstrate no cytotoxic effects on the iPS as well as an increased expression of cardiac markers for both stimulated and unstimulated protocols. This study demonstrates the first application of PPy as a supportive electroactive material for iPS and the first development of a fiber scaffold capable of dynamic mechanical actuation.

Biocompatibility of Polypyrrole with Human Primary Osteoblasts and the Effect of Dopants.

Polypyrrole (PPy) is a conducting polymer that enables controlled drug release upon electrical stimulation. We characterized the biocompatibility of PPy with human primary osteoblasts, and the effect of dopants. We investigated the biocompatibility of PPy comprising various dopants, i.e. p-toluene sulfonate (PPy-pTS), chondroitin sulfate (PPy-CS), or dodecylbenzenesulfonate (PPy-DBS), with human primary osteoblasts. PPy-DBS showed the roughest appearance of all surfaces tested, and its wettability was similar to the gold-coated control. The average number of attached cells was 45% higher on PPy-DBS than on PPy-CS or PPy-pTS, although gene expression of the proliferation marker Ki-67 was similar in osteoblasts on all surfaces tested. Osteoblasts seeded on PPy-DBS or gold showed similar vinculin attachment points, vinculin area per cell area, actin filament structure, and Feret's diameter, while cells seeded on PPY-CS or PPY-pTS showed disturbed focal adhesions and were enlarged with disorganized actin filaments. Osteoblasts grown on PPy-DBS or gold showed enhanced alkaline phosphatase activity and osteocalcin gene expression, but reduced osteopontin gene expression compared to cells grown on PPy-pTS and PPy-CS. In conclusion, PPy doped with DBS showed excellent biocompatibility, which resulted in maintaining focal adhesions, cell morphology, cell number, alkaline phosphatase activity, and osteocalcin gene expression. Taken together, conducting polymers doped with DBS are well tolerated by osteoblasts. Our results could provide a basis for the development of novel orthopedic or dental implants with controlled release of antibiotics and pharmaceutics that fight infections or focally enhance bone formation in a tightly controlled manner.

Electronic control of platelet adhesion using conducting polymer microarrays.

We hereby report a method to fabricate addressable micropatterns of e-surfaces based on the conducting polymer poly(3,4-ethylenedioxythiophene) doped with the anion tosylate (PEDOT:Tos) to gain dynamic control over the spatial distribution of platelets in vitro. With thin film processing and microfabrication techniques, patterns down to 10 µm were produced to enable active regulation of platelet adhesion at high spatial resolution. Upon electronic addressing, both reduced and oxidized surfaces were created within the same device. This surface modulation dictates the conformation and/or orientation, rather than the concentration, of surface proteins, thus indirectly regulating the adhesion of platelets. The reduced electrode supported platelet adhesion, whereas the oxidized counterpart inhibited adhesion. PEDOT:Tos electrode fabrication is compatible with most of the classical patterning techniques used in printing as well as in the electronics industry. The first types of tools promise ultra-low-cost production of low-resolution (>30 µm) electrode patterns that may combine with traditional substrates and dishes used in a classical analysis setup. Platelets play a pronounced role in cardiovascular diseases and have become an important drug target in order to prevent thrombosis. This clinical path has in turn generated a need for platelet function tests to monitor and assess platelet drug efficacy. The spatial control of platelet adherence presented here could prove valuable for blood cell separation or biosensor microarrays, e.g. in diagnostic applications where platelet function is evaluated.