論文 - 尾上　弘晃

Controlling Firing Point of Microfiber-Shaped Hipsc-Derived Cardiac Tissue with Localized Electrical Stimulation Device

Masuda A., Itai S., Kurashina Y., Tohyama S., Onoe H.

Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS) （Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)）  2023-January   305 - 308 2023年

ISSN  10846999

　概要を見る

We propose a device to apply localized electrical stimulation to control cardiac conduction in in vitro cardiac tissue. Our system can initiate the generation of electrical impulses at any location by applying local electrical stimulation to the desired point of the microfiber-shaped cardiac tissue. Our system generated a strong and intense electric field between the electrodes in FEM analysis, indicating only the cardiac tissue between the electrodes was electrically stimulated. We could observe the cardiac conduction optically by visualizing the timing and location of impulses with calcium imaging at any location and time on the tissue. The system enables accurate analysis of cardiac potential and conduction velocity of the heart model, and could contribute to the elucidation of pathological dynamics such as arrhythmia, whose mechanism of occurrence still remains unknown.

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A Three-Dimensional Artificial Intestinal Tissue With A Crypt-Like Inner Surface

Tanaka S., Itai S., Onoe H.

Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS) （Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)）  2023-January   456 - 459 2023年

ISSN  10846999

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Disruption of the microbiome in humans caused several diseases. To infer causality, the intestinal models are essential, but the commonly used model has still room for improvements such as biomimicry, productivity, and ability to co-culture with bacteria. This paper describes the artificial intestinal tissue by culturing intestinal cells (Caco-2) in the collagen gel tube with crypt-like cavities created by electrolysis-triggered microbubbles. This tube device has high biomimicry and productivity and also consists of glass/silicone tubes connector at both ends, which allows for connecting external pumps easily and running two types of flows: the inside of the tube (internal flow) and permeation from the outside of the tube to the inside (external flow). In order to show this high extensibility, we demonstrated the co-culture of Caco-2 cells and bacteria (Bifidobacteria). We believe that our device will be a platform for investigating complex intestinal diseases due to its high biomimetic properties.

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Repeatable detection of Ag⁺ ions using a DNA aptamer-linked hydrogel biochemical sensor integrated with microfluidic heating system

Yoshida K., Hayashi T., Takinoue M., Onoe H.

Scientific Reports （Scientific Reports）  12 （ 1 ）  2022年12月

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This paper describes repeatable detection of Ag+ ions using a DNA aptamer-linked hydrogel biochemical sensor integrated with a microfluidic heating system. Biochemical sensors that respond to chemical compounds and produce detectable signals have a critical role in many aspects of modern society. In particular, the repeatable measurement of environmental information such as toxic substances including Ag+ ions could be expected to improve the environment. The DNA aptamer is an attractive candidate because of the stability and the selectivity of binding to chemicals. However, previous DNA aptamer biochemical sensors could not measure repeatedly because those sensors did not have initializing functions. To overcome this challenge, we proposed a DNA aptamer-linked hydrogel biochemical sensor integrated with the microfluidic heating system enabling repeatable detection of Ag+ ions. The binding Ag+ ions are dissociated by heating and flushing through the integrated microfluidic heating device. The DNA aptamer-linked hydrogel had the capability to detect a wide range of Ag+ ion concentrations (10−5–10 mM) including a toxic range for various aquatic organisms. Finally, we demonstrated the repeatable detection of the Ag+ ions. These results indicated that our proposed biochemical sensor is expected to use for long-term monitoring with high stability in ambient temperature and low power consumption.

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Light-driven flow control in the hydrogel microchannel by utilizing a metal microstructure fabricated by multiphoton photoreduction

Nagano Y., Onoe H., Terakawa M.

Optical Materials Express （Optical Materials Express）  12 （ 8 ） 3303 - 3312 2022年08月

　概要を見る

The light-stimulated control of flow velocity in the hydrogel microchannel was demonstrated by utilizing a metal microstructure fabricated by multiphoton photoreduction. The metal microstructure was fabricated adjacent to the microchannel in a poly-N- isopropylacrylamide (PNIPAm) hydrogel. Owing to the photothermal conversion at the metal microstructure, local deformation of the microchannel and change in flow velocity was induced as a result of the volume phase transition of the supporting hydrogel around the metal microstructure. Wavelength-selective change in the flow velocity was realized by utilizing dissimilar metal microstructures, that exhibit different optical resonances. The results indicate that multiphoton photoreduction is a promising method for fabricating novel hydrogel devices having flow-controllable switches by arranging light-absorbing structures with high spatial selectivity.

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Vascular embolization of radiopaque hydrogel microfiber using ultra-minimally invasive technique for stage-adjustable renal failure model

Takakura N., Ohta H., Komatsu T., Kurashina Y., Hiroka Y., Okano H.J., Onoe H.

Materials and Design （Materials and Design）  219 2022年07月

ISSN  02641275

　概要を見る

Animal pathology models play an important role in elucidating pathology, evaluating drug efficacy, and studying therapeutic. This paper presents the creation of rats’ renal failure model by embolizing the renal artery using radiopaque hydrogel microfibers. By using a dual coaxial laminar flow microfluidic device, barium alginate gel microfibers containing zirconia particles were fabricated. Since the zirconia particles are radiopaque, the microfiber can be delivered and embolized while confirming the position of the microfiber in real-time under X-ray imaging. The delivery of microfibers through a catheter into the renal artery of rats to create a renal failure model was tested. The radiopaque microfibers were visualized by X-ray imaging when delivered by a catheter. Furthermore, the microfibers remained stable for 2 weeks after delivery. In addition, by adjusting the number of microfibers, two different rat models with severe and mild renal failure conditions were created. The proposed delivery of radiopaque hydrogel microfibers to create an embolization model is expected to be an effective approach to control the degree of symptoms and elucidate the pathology.

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A novel fabrication process of up-scalable microfiber-shaped tendon-like tissue with high cell density for uniformed macroscale assembly

Fukada K., Tachibana K., Kurashina Y., Kaneko Y., Matsumoto T., Miyamoto T., Niki Y., Nakamura M., Onoe H.

Biotechnology and Bioengineering （Biotechnology and Bioengineering）  119 （ 5 ） 1327 - 1336 2022年05月

ISSN  00063592

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This paper describes up-scalable microfiber-shaped tissues for macroscale tendon tissue reconstruction in vitro. C3H10T1/2 cells were encapsulated in a calcium alginate hydrogel microfiber that was fabricated via a double coaxial microfluidic device. The C3H10T1/2 cells gradually merged to construct the microfiber-shaped tendon-like tissue. Our microfiber-shaped tendon-like tissues were alive and maintained their microfiber-shaped morphology over 600 days. Immunostaining and real-time quantitative polymerase chain reaction analyses showed that our fabricated microfiber-shaped tendon-like tissue properly expressed tenomodulin and the orientation of the filaments of actin, which are one of the characteristics of tendon tissue in vivo. Furthermore, a macroscale tendon tissue assembly with ∼1 cm in length and ∼200 µm in thickness was successfully constructed by bundling the microfiber-shaped tendon-like tissues together. This feature enabled us to fabricate a macroscale tendon tissue with uniform cell distribution. We believe that our fabricated microfiber-shaped tendon-like tissue would be a suitable strategy to reconstruct tendon tissue in vitro for the treatments of tendon-related injuries.

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Microfiber-Shaped Programmable Materials with Stimuli-Responsive Hydrogel

Takeuchi N., Nakajima S., Yoshida K., Kawano R., Hori Y., Onoe H.

Soft Robotics （Soft Robotics）  9 （ 1 ） 89 - 97 2022年02月

ISSN  21695172

　概要を見る

Programmable materials have artificially designed physical shapes responding to external stimuli, as well as high design capability and high flexibility. Here, we propose a microfiber-shaped programmable material with an axial pattern of stimuli-responsive (SR) and nonresponsive hydrogels. The SR pre-gel solution was mixed to sodium alginate pre-gel solution for instantaneous gelation with ionic crosslinking and solidified on a nonresponsive hydrogel microfiber with a valve-controlled microfluidic system. A design of microfiber-shaped programmable material (patterned position of SR regions) could be flexibly altered by changing a coded sequence program. We confirmed that the three-dimensional (3D) coil-like structures were self-folded at the patterned SR regions responding to the thermal stimulus and that the chirality of the self-folded 3D coil-like structures depends on the condition of the stimulus to the microfiber. Finally, interaction with objects using the programmable microfiber as a soft actuator was demonstrated. Our microfiber-shaped programmable materials expand possibilities of fiber-based materials in biomimetics and soft robotics fields.

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Microfluidic multicolor display by juxtapositional color mixing with a pattern of primary color pixels

Muramatsu J., Onoe H.

Journal of Micromechanics and Microengineering （Journal of Micromechanics and Microengineering）  32 （ 2 ）  2022年02月

ISSN  09601317

　概要を見る

This paper describes a microfluidic multicolor display utilizing juxtapositional color mixing of pixels. Our display has a 14 14 array of pixels (2.5 mm in pixel diameter, 8.46 ppi) on the display surface where multicolor is expressed by controlling the pattern of the four primary color inks (cyan (C), magenta (M), yellow (Y), and black (= key plate, K)) that fill the pixels. The microfluidic display has a three-layer structure composed of a top layer with pixels for displaying images, a middle layer that serves as a background screen, and a bottom layer with microchannels that connect the pixels. In order to express multicolor by combining CMYK primary colors, we optimized the concentration of the inks used as the primary colors. By designing patterns of pixels filled with CMYK ink, color gradations and multicolor images were displayed on our display. The proposed microfluidic display could be applied to eye-friendly and low-energy-consumption flexible display applications including multi-purpose sign boards used in outdoors, wearing objects, and exterior/interior of vehicles and architects.

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Flexibly Deformable Collagen Hydrogel Tube Reproducing Immunological Tissue Deformation of Blood Vessels as a Pharmacokinetic Testing Model

Itai S., Onoe H.

Advanced Healthcare Materials （Advanced Healthcare Materials）  11 （ 1 ）  2022年01月

ISSN  21922640

　概要を見る

Since the biochemical reaction of blood vessels plays an essential role in immune response or various diseases, in vitro vascular models have high demand from medical fields. However, vascular models often tend to be difficult to mimic the biomedical reaction faithfully because of the lack of implementation of the tissue deformation. Here, an inflammatory mediator-induced deformation reaction of a blood vessel on a flexibly deformable collagen hydrogel tube device is reproduced. A self-standing collagen tube enables the tissue to deform flexibly in biochemical reaction and achieves contraction both at tissue and cell level. The contraction of tissue relieves the stress between cells under reaction to maintain cellular junctions even tight junctions are broken. Also, the drug perfusion test with antihistamine chemical is easily performed due to the connector part and properly inhibits the inflammatory reaction. Moreover, the traction force on endothelial cells is analyzed as about 0.9 µN on two types of scaffolds with different stiffness. It is believed that the potential of the flexible tissue model to reproduce biochemical reactions can contribute to the fabrication of vascular tissue models mimicking in vivo in high similarity as a platform for biomedical researches and pharmacokinetic testing.

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Fixation-Free Evaluation of Cardiac Contractile Force by Human iPSC-Derived Cardiac Core-Shell Microfiber

Masuda A., Fukada K., Itai S., Kurashina Y., Akizuki S., Tohyama S., Fujita J., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   172 - 175 2022年

ISSN  21601968

　概要を見る

We propose a core-shell microfiber-shaped cardiac tissue as a drug screening model with a fixation-free contractile force measurement system. The contractile force of the hiPSC-derived cardiac tissue can be calculated only with a motion analysis, thanks to a shell of hydrogel covering the cardiac tissue. Thus, the system can evaluate the behavior of the tissue in drug screenings without any specific equipment. We demonstrated drug screening tests with pharmaceuticals on our microfiber tissues, resulting in 102.7% increase of contractile force with isoproterenol and 56.3% decrease with propranolol. Our cardiac drug screening model can easily reproduce and evaluate cardiac disease models, and would contribute to the pathological research of unexplained cardiac diseases and pharmacokinetic testing.

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Tube-Shaped In-Vitro Intestinal Gut Model with 3D Isotropic Medium Supply for Bacterial Symbiosis

Kawahara M., Itai S., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   321 - 324 2022年

ISSN  21601968

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This paper describes a tube-shaped in vitro intestinal gut model achieving both isotropic medium supply and stable co-culture with bacteria. Our tube-shaped device consists of a collagen/Matrigel tube as a scaffold for intestinal cells and glass/silicone tubes at both ends for perfusion. We confirmed intestinal cells (Caco-2) can be maintained in the luminal part with nutrient-free buffer flow mimicking bacterial suspensions and demonstrated the co-culture of Escherichia-coli (E. coli) in the Caco-2 cultured tube-shaped device. We believe this device provides ple but stable co-culture model with intestinal cells and bacteria and contributes to medical field research relating intestinal bacteria.

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Protein Expression Micro-Scale Mapping Analysis of Endothelial Tissue in in Vitro Branched Vascular Model under Mechanical Stimuli

Muramatsu J., Shimizu A., Hashimoto M., Miura S., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   283 - 286 2022年

ISSN  21601968

　概要を見る

This paper describes a micro-scale mapping of protein expression on endothelial tissue in an extracellular matrix (ECM)-based branched microchannel under mechanical stimuli. Our system mimics the heterogeneity in the distribution of complex mechanical stimuli applied to tissues in vivo by applying mechanical stimuli to a branched vascular model. We confirmed the up-regulation of gene expression levels in vascular endothelial cells(ECs) by stretch stress. We cultured ECs under complex mechanical stimuli, and visualized the distribution of mechanotransducer protein expression in the branched vascular model. We believe that our system could open a new door for the analysis of local cellular responses to mechanical stimuli.

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Near Infrared-Triggered On-Demand Adeno-Associated Virus Release from Hydrogel Microbeads for Gene Therapy

Takatsuka S., Kubota T., Kurashina Y., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   388 - 391 2022年

ISSN  21601968

　概要を見る

This paper describes near infrared (NIR)-triggered on-demand adeno-associated virus (AAV) release from alginate hydrogel microbeads with heat transducer for gene therapy. As an AAV carrier, calcium alginate (Ca-Alg.) hydrogel microbeads were fabricated by injecting mixed pre-gel solution composed of sodium-alginate (Na-Alg.), AAV, and Fe3O4 particles (FPs: heat transducer) to CaCl2 solution to form ionically cross-linked instantly with a centrifuge-based microdroplet shooting device. The FPs generate heat due to localized surface plasmon resonance under NIR. By irradiating NIR to the microbeads, we confirmed the release of AAV and the gene transfection to HeLa cells. Our proposed NIR-triggered AAV release system would be an effective and safe controlled release system for gene therapy.

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Mechanically Magnified Sensitivity Enhancement with Micro-Step Substrate for Eye-Recognizable Structural-Color Hydrogel Biochemical Sensors

Yamawaki S., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   739 - 742 2022年

ISSN  21601968

　概要を見る

We present a versatile device structure that can mechanically magnify the sensitivity of stimuli-responsive structural-color hydrogel biochemical sensors. In our system, the structural-color hydrogel part and the stimuli-response hydrogel part are separately arrayed on a substrate with micro-step structures, causing the height difference between the two hydrogel parts. This height difference magnifies the shrinkage ratio to be larger in the color part than in the stimulus-responsive part, achieving the x5 - x10 high sensitivity of the hydrogel-based biochemical sensor. We applied ethanol-responsive and temperature-responsive hydrogels to our proposed device and confirmed the magnification of the sensitivity compared to previous sensors. We believe that our sensor device will be an effective approach to providing sensing with practical sensitivity for medical and other applications.

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Ingestible Wireless Capsule Sensor Made from Edible Materials for Gut Bacteria Monitoring

Inami A., Kan T., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   110 - 113 2022年

ISSN  21601968

　概要を見る

This report describes an ingestible capsule sensor that can wirelessly detect the activity of gut bacteria by irradiating microwaves from outside the body. The proposed sensor is composed of only edible materials: the critical sensing material is a dietary fiber, guar gum, that is decomposed by gut bacteria. We confirmed that the guar gum film can be decomposed by gut bacteria, bifidobacteria. For wireless communication, we fabricated an ingestible antenna made from rice paper and gold with a split-ring resonator (SRR), one of the elements of electromagnetic metamaterials. Our sensor fulfills swallowable size and wireless communication requirements for the first time and would be a safety sensor for gastrointestinal (GI) tract monitoring. We believe that our ingestible sensor could be an effective tool for non-invasive biological substances monitoring without retention risk.

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In Vitro Artery Model with Circumferentially Aligned & Contractible Smooth Muscle by Unfixed Molding & Screwing Fabrication

Itai S., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   275 - 278 2022年

ISSN  21601968

　概要を見る

This paper describes a fabrication method of an artery model with circumferentially aligned and contractible smooth muscle cells (SMCs) by unfixed molding and screwing processes. The fabricated aligned smooth muscle is well-deformable, enabling the flexible contraction in biological reaction. The axially aligned fiber-shaped smooth muscle tissue was formed by unfixed molding and screwed up to be aligned circumferentially. Also, the SMC layer was covered with a collagen layer with connectors to enable co-culture with endothelial cells (ECs) and perfusion culture. The alignment of SMCs was visibly confirmed, and the ability to reproduce the biochemical contraction was proved by an inflammatory reaction. We believe that our novel fabrication method would contribute to the development of functional smooth muscle models for biomedical researches.

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Glass-Capillary-Embedded 3D Coaxial Microfluidic Device with Pneumatic Microvalve Control for Producing Patterned Functional Materials

Takakura N., Kurashina Y., Onoe H.

IEEE Symposium on Mass Storage Systems and Technologies （IEEE Symposium on Mass Storage Systems and Technologies）  2022-January   267 - 270 2022年

ISSN  21601968

　概要を見る

This paper describes a glass-embedded poly-dimethyl-siloxane microfluidic device with integrated microvalves to create a patterned 3D coaxial flow for producing functional composite materials. The microfluidic device was fabricated by transferring two layers of PDMS onto acrylic molds and bonding them across a glass capillary. We confirmed that the microfluidic device could flow coaxial flow and switch the solution. We demonstrated that core-shell microfibers with shells patterned in two different directions by flowing three different sodium alginate solutions into the microfluidic device. Our proposed microfluidic device would be used to fabricate multifunctional hydrogel microfibers made of multiple materials, which can be applied to bio-fabrication and soft-actuator.

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WIRELESS SOIL PH SENSING IN OUTDOOR ENVIRONMENT WITH PH-SELECTIVE FULLY-DEGRADABLE METAMATERIAL ANTENNA

Sakabe K., Kan T., Onoe H.

MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences （MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences）     217 - 218 2022年

　概要を見る

We propose a wireless and battery-free soil pH sensor made entirely of degradable materials for soil pH monitoring in outdoor environments. The sensor consists of magnesium (Mg) electric antenna based on inductor-capacitor circuit resonance with hydroxyapatite (Ca10(PO4)6(OH)2, HA) coating, enabling degradation of the sensor in acid soil conditions. In this report, we succeeded in measuring the changes over time of different electromagnetic responses in soils with different soil pH. Furthermore, the electromagnetic response of the sensor was also demonstrated in the outdoor field experiment. We believe that our biodegradable soil pH sensor could apply to low-cost, in situ, and environmentally friendly monitoring of soil pH.

A THREE-DIMENSIONAL ARTIFICIAL INTESTINAL TUBE WITH A CRYPT-LIKE INNER SURFACE FORMED BY ELECTROLYTIC MICROBUBBLES

Tanaka S., Itai S., Onoe H.

MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences （MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences）     13 - 14 2022年

　概要を見る

Crypts-like structures are indispensable to fabricating an artificial intestinal tissue reproducing the arrangement of cell types in vivo. This paper presents fabrication of an artificial intestinal tube with an uneven inner wall created by electrolysis-triggered microbubbles. During the fabrication process of a scaffold, microbubbles were generated on a tungsten electrode to form cavities on the inner surface of the collagen hydrogel tube. The method can fabricate numerous crypt-like structures in the order of a hundred micro-meter quickly. We believe that our device will be a platform for investigating complex intestinal diseases due to its high biomimetic properties.

L-2L LADDER MICROFLUIDICS FOR DYNAMIC GENERATION OF CHEMICAL CONCENTRATIONS

Shiraishi S., Chatani T., Miyazako H., Onoe H., Hori Y.

MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences （MicroTAS 2022 - 26th International Conference on Miniaturized Systems for Chemistry and Life Sciences）     897 - 898 2022年

　概要を見る

System-level understanding of dynamic biological process requires the measurement of the dynamic response of cells to chemical stimulations with various waveforms. Here, we develop a simple and scalable microfluidic chemical stimulator capable of modulating the concentration waveforms. The proposed device is based on the analogy of an electronic digital-analog converter (DAC), and thereby, the waveforms can be controlled by an intuitive binary logic. We demonstrate dynamic modulation of solution concentrations and perform systematic characterization of the waveforms to enable facile tuning and scale-up of the device.