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1. Biocompatible micro tweezers for 3D hydrogel organoid array mechanical characterization

   https://doi.org/10.1371/journal.pone.0262950  

   Alhudaithy, Soliman ; Hoshino, Kazunori ( January 2022 , PLOS ONE)

   Jabbari, Esmaiel (Ed.)

   This study presents novel biocompatible Polydimethylsiloxane (PDMS)-based micromechanical tweezers (μTweezers) capable of the stiffness characterization and manipulation of hydrogel-based organoids. The system showed great potential for complementing established mechanical characterization methods such as Atomic Force Microscopy (AFM), parallel plate compression (PPC), and nanoindentation, while significantly reducing the volume of valuable hydrogels used for testing. We achieved a volume reduction of ~0.22 μl/sample using the μTweezers vs. ~157 μl/sample using the PPC, while targeting high-throughput measurement of widely adopted micro-mesoscale (a few hundred μm-1500 μm) 3D cell cultures. The μTweezers applied and measured nano-millinewton forces through cantilever’ deflection with high linearity and tunability for different applications; the assembly is compatible with typical inverted optical microscopes and fit on standard tissue culture Petri dishes, allowing mechanical compression characterization of arrayed 3D hydrogel-based organoids in a high throughput manner. The average achievable output per group was 40 tests per hour, where 20 organoids and 20 reference images in one 35 mm petri dish were tested, illustrating efficient productivity to match the increasing demand on 3D organoids’ applications. The changes in stiffness of collagen I hydrogel organoids in four conditions were measured, with ovarian cancer cells (SKOV3) or without (control). The Young’s modulus ofmore »the control group (Control—day 0, E = 407± 146, n = 4) measured by PPC was used as a reference modulus, where the relative elastic compressive modulus of the other groups based on the stiffness measurements was also calculated (control-day 0, E = 407 Pa), (SKOV3-day 0, E = 318 Pa), (control-day 5, E = 528 Pa), and (SKOV3-day 5, E = 376 Pa). The SKOV3-embedded hydrogel-based organoids had more shrinkage and lowered moduli on day 0 and day 5 than controls, consistently, while SKOV3 embedded organoids increased in stiffness in a similar trend to the collagen I control from day 0 to day 5. The proposed method can contribute to the biomedical, biochemical, and regenerative engineering fields, where bulk mechanical characterization is of interest. The μTweezers will also provide attractive design and application concepts to soft membrane-micro 3D robotics, sensors, and actuators.« less
2. Autonomic perspiration in 3D-printed hydrogel actuators

   https://doi.org/10.1126/scirobotics.aaz3918  

   Mishra, Anand K. ; Wallin, Thomas J. ; Pan, Wenyang ; Xu, Patricia ; Wang, Kaiyang ; Giannelis, Emmanuel P. ; Mazzolai, Barbara ; Shepherd, Robert F. ( January 2020 , Science Robotics)

   In both biological and engineered systems, functioning at peak power output for prolonged periods of time requires thermoregulation. Here, we report a soft hydrogel-based actuator that can maintain stable body temperatures via autonomic perspiration. Using multimaterial stereolithography, we three-dimensionally print finger-like fluidic elastomer actuators having a poly- N -isopropylacrylamide (PNIPAm) body capped with a microporous (~200 micrometers) polyacrylamide (PAAm) dorsal layer. The chemomechanical response of these hydrogel materials is such that, at low temperatures (<30°C), the pores are sufficiently closed to allow for pressurization and actuation, whereas at elevated temperatures (>30°C), the pores dilate to enable localized perspiration in the hydraulic actuator. Such sweating actuators exhibit a 600% enhancement in cooling rate (i.e., 39.1°C minute −1 ) over similar non-sweating devices. Combining multiple finger actuators into a single device yields soft robotic grippers capable of both mechanically and thermally manipulating various heated objects. The measured thermoregulatory performance of these sweating actuators (~107 watts kilogram −1 ) greatly exceeds the evaporative cooling capacity found in the best animal systems (~35 watts kilogram −1 ) at the cost of a temporary decrease in actuation efficiency.
3. Poly(alkyl glycidyl ether) hydrogels for harnessing the bioactivity of engineered microbes

   https://doi.org/10.1039/c9fd00019d  

   Johnston, Trevor G. ; Fellin, Christopher R. ; Carignano, Alberto ; Nelson, Alshakim ( October 2019 , Faraday Discussions)

   Herein, we describe a method to produce yeast-laden hydrogel inks for the direct-write 3D printing of cuboidal lattices for immobilized whole-cell catalysis. A poly(alkyl glycidyl ether)-based triblock copolymer was designed to have three important features for this application: (1) a temperature response, which allowed for facile processing of the material; (2) a shear response, which facilitated the extrusion of the material through a nozzle; and (3) UV light induced polymerization, which enabled the post-extrusion chemical crosslinking of network chains, and the fabrication of robust printed objects. These three key stimuli responses were confirmed via rheometrical characterization. A genetically-engineered yeast strain with an upregulated α-factor production pathway was incorporated into the hydrogel ink and 3D printed. The immobilized yeast cells exhibited adequate viability of 87.5% within the hydrogel. The production of the upregulated α-factor was detected using a detecting yeast strain and quantified at 268 nM ( s = 34.6 nM) over 72 h. The reusability of these bioreactors was demonstrated via immersion of the yeast-laden hydrogel lattice in fresh SC media and confirmed by the detection of similar amounts of upregulated α-factor at 259 nM ( s = 45.1 nM). These yeast-laden materials represent an attractive opportunity for whole-cell catalysismore »of other high-value products in a sustainable and continuous manner.« less
4. 3D Printing Hydrogel-Based Soft and Biohybrid Actuators: A Mini-Review on Fabrication Techniques, Applications, and Challenges

   https://doi.org/10.3389/frobt.2021.673533  

   Sun, Wenhuan ; Schaffer, Saul ; Dai, Kevin ; Yao, Lining ; Feinberg, Adam ; Webster-Wood, Victoria ( April 2021 , Frontiers in Robotics and AI)

   Stimuli-responsive hydrogels are candidate building blocks for soft robotic applications due to many of their unique properties, including tunable mechanical properties and biocompatibility. Over the past decade, there has been significant progress in developing soft and biohybrid actuators using naturally occurring and synthetic hydrogels to address the increasing demands for machines capable of interacting with fragile biological systems. Recent advancements in three-dimensional (3D) printing technology, either as a standalone manufacturing process or integrated with traditional fabrication techniques, have enabled the development of hydrogel-based actuators with on-demand geometry and actuation modalities. This mini-review surveys existing research efforts to inspire the development of novel fabrication techniques using hydrogel building blocks and identify potential future directions. In this article, existing 3D fabrication techniques for hydrogel actuators are first examined. Next, existing actuation mechanisms, including pneumatic, hydraulic, ionic, dehydration-rehydration, and cell-powered actuation, are reviewed with their benefits and limitations discussed. Subsequently, the applications of hydrogel-based actuators, including compliant handling of fragile items, micro-swimmers, wearable devices, and origami structures, are described. Finally, challenges in fabricating functional actuators using existing techniques are discussed.
5. Long-Fiber Embedded Hydrogel 3D Printing for Structural Reinforcement

   https://doi.org/10.1021/acsbiomaterials.1c00908  

   Sun, Wenhuan ; Tashman, Joshua W. ; Shiwarski, Daniel J. ; Feinberg, Adam W. ; Webster-Wood, Victoria A. ( December 2021 , ACS Biomaterials Science & Engineering)

   Hydrogels are candidate building blocks in a wide range of biomaterial applications including soft and biohybrid robotics, microfluidics, and tissue engineering. Recent advances in embedded 3D printing have broadened the design space accessible with hydrogel additive manufacturing. Specifically, the Freeform Reversible Embedding of Suspended Hydrogels (FRESH) technique has enabled the fabrication of complex 3D structures using extremely soft hydrogels, e.g., alginate and collagen, by assembling hydrogels within a fugitive support bath. However, the low structural rigidity of FRESH printed hydrogels limits their applications, especially those that require operation in nonaqueous environments. In this study, we demonstrated long-fiber embedded hydrogel 3D printing using a multihead printing platform consisting of a custom-built fiber extruder and an open-source FRESH bioprinter with high embedding fidelity. Using this process, fibers were embedded in 3D printed hydrogel components to achieve significant structural reinforcement (e.g., tensile modulus improved from 56.78 ± 8.76 to 382.55 ± 25.29 kPa and tensile strength improved from 9.44 ± 2.28 to 45.05 ± 5.53 kPa). In addition, we demonstrated the versatility of this technique by using fibers of a wide range of sizes and material types and implementing different 2D and 3D embedding patterns, such as embedding a conical helix using electrochemicallymore »aligned collagen fiber via nonplanar printing. Moreover, the technique was implemented using low-cost material and is compatible with open-source software and hardware, which facilitates its adoption and modification for new research applications.« less