Extracellular Microenvironmental Control for Organoid Assembly.

Organoids, which are multicellular clusters with similar physiological functions to living organs, have gained increasing attention in bioengineering. As organoids become more advanced, methods to form complex structures continue to develop. There is evidence that the extracellular microenvironment can regulate organoid quality. The extracellular microenvironment consists of soluble bioactive molecules, extracellular matrix, and biofluid flow. However, few efforts have been made to discuss the microenvironment optimal to engineer specific organoids. Therefore, this review article examines the extent to which engineered extracellular microenvironments regulate organoid quality. First, we summarize the natural tissue and organ's unique chemical and mechanical properties, guiding researchers to design an extracellular microenvironment used for organoid engineering. Then, we summarize how the microenvironments contribute to the formation and growth of the brain, lung, intestine, liver, retinal, and kidney organoids. The approaches to forming and evaluating the resulting organoids are also discussed in detail. Impact statement Organoids, which are multicellular clusters with similar physiological function to living organs, have been gaining increasing attention in bioengineering. As organoids become more advanced, methods to form complex structures continue to develop. This review article focuses on recent efforts to engineer the extracellular microenvironment in organoid research. We summarized the natural organ's microenvironment, which informs researchers of key factors that can influence organoid formation. Then, we summarize how these microenvironmental controls significantly contribute to the formation and growth of the corresponding brain, lung, intestine, liver, retinal, and kidney organoids. The approaches to forming and evaluating the resulting organoids are discussed in detail, including extracellular matrix choice and properties, culture methods, and the evaluation of the morphology and functionality through imaging and biochemical analysis.

Electrothermal soft manipulator enabling safe transport and handling of thin cell/tissue sheets and bioelectronic devices.

"Living" cell sheets or bioelectronic chips have great potentials to improve the quality of diagnostics and therapies. However, handling these thin and delicate materials remains a grand challenge because the external force applied for gripping and releasing can easily deform or damage the materials. This study presents a soft manipulator that can manipulate and transport cell/tissue sheets and ultrathin wearable biosensing devices seamlessly by recapitulating how a cephalopod's suction cup works. The soft manipulator consists of an ultrafast thermo-responsive, microchanneled hydrogel layer with tissue-like softness and an electric heater layer. The electric current to the manipulator drives microchannels of the gel to shrink/expand and results in a pressure change through the microchannels. The manipulator can lift/detach an object within 10 s and can be used repeatedly over 50 times. This soft manipulator would be highly useful for safe and reliable assembly and implantation of therapeutic cell/tissue sheets and biosensing devices.

Preoperative vascular surgery model using a single polymer tough hydrogel with controllable elastic moduli.

Materials used in organ mimics for medial simulation and education require tissue-like softness, toughness, and hydration to give clinicians and students accurate tactile feedback. However, there is a lack of materials that satisfy these requirements. Herein, we demonstrate that a stretchable and tough polyacrylamide hydrogel is useful to build organ mimics that match softness, crack growth resistance, and interstitial water of real organs. Varying the acrylamide concentration between 29 or 62% w/w with a molar ratio between cross-linker and acrylamide of 1 : 10 800 resulted in a fracture energy around ∼2000 J m-2. More interestingly, this tough gel permitted variation of the elastic modulus from 8 to 62 kPa, which matches the softness of brain to vascular and muscle tissue. According to the rheological frequency sweep, the tough polyacrylamide hydrogels had a greatly decreased number of flow units, indicating that when deformed, stress was dispersed over a greater area. We propose that such molecular dissipation results from the increased number of entangled polymers between distant covalent cross-links. The gel was able to undergo various manipulations including stretching, puncture, delivery through a syringe tip, and suturing, thus enabling the use of the gel as a blood vessel model for microsurgery simulation.

Transparent and Flexible Electronics Assembled with Metallic Nanowire-Layered Nondrying Glycerogel.

There has been increasing demand for transparent and mechanically durable electrical conductors for their uses in wearable electronic devices. It is common to layer metallic nanowires on transparent but stiff poly(dimethylsiloxane) (PDMS) or stretchable but opaque Ecoflex-based substrates. Here, we hypothesized that layering metallic nanowires on a stretchable and hygroscopic gel would allow us to assemble a transparent, stretchable, and durable conductor. The hygroscopic property of the gel was attained by partially replacing water in the preformed polyacrylamide hydrogel with glycerol. The resulting gel, denoted as a glycerogel, could remain hydrated for over 6 months in air by taking up water molecules from the air. The glycerogel was tailored to be stretchable up to 8 times its original length by tuning the amount of the cross-linker and acrylamide. The resulting glycerogel allowed for deposition of wavy silver nanowires using the prestrain method up to 400% prestrain, without causing kinks and interfacial cracks often found with nanowires layered onto PDMS. With a prestrain of 100%, the resulting nanowire-gel conductor exhibited optical transparency (85%) and electrical conductivity (17.1 ohm/sq) even after 5000 cycles of deformation. The results of this study would broadly be useful to improve the performance of the next generation of flexible electronic devices.

Reactive oxygen species-responsive drug delivery systems for the treatment of neurodegenerative diseases.

Neurodegenerative diseases and disorders seriously impact memory and cognition and can become life-threatening. Current medical techniques attempt to combat these detrimental effects mainly through the administration of neuromedicine. However, drug efficacy is limited by rapid dispersal of the drugs to off-target sites while the site of administration is prone to overdose. Many neuropathological conditions are accompanied by excessive reactive oxygen species (ROS) due to the inflammatory response. Accordingly, ROS-responsive drug delivery systems have emerged as a promising solution. To guide intelligent and comprehensive design of ROS-responsive drug delivery systems, this review article discusses the two following topics: (1) the biology of ROS in both healthy and diseased nervous systems and (2) recent developments in ROS-responsive, drug delivery system design. Overall, this review article would assist efforts to make better decisions about designing ROS-responsive, neural drug delivery systems, including the selection of ROS-responsive functional groups.

Stretchable, anti-bacterial hydrogel activated by large mechanical deformation.

Hydrogels have been used extensively to deliver functional molecular cargos in response to external mechanical force. However, the intrinsic brittleness of gels restricts the applicable range of strain to 0.1, thus limiting the range of molecular release rate that may be controlled. Also, uncontrollable molecular diffusion, which is especially prominent in small molecules, reduces the role of mechanical stimulus on the release rate. As such, we hypothesized that these challenges would be resolved by combining cyclodextrin, which may form guest-host complexes with small molecular cargos, with a stretchable hydrogel system. We examined this hypothesis by synthesizing cyclodextrin acrylate and incorporating it into a polyacrylamide gel that can be stretched by 100% of its original length. In the absence of external stretching, hydrogels containing cyclodextrin acrylate with a degree of acryloyl group substitution (DSA) of 2.3 presented a lower molecular release rate than hydrogels without cyclodextrin acrylate. More interestingly, the polyacrylamide-cyclodextrin hydrogel system displayed an increased molecular release rate corresponding to the degree of stretching, particularly in the gels containing cyclodextrin acrylate with a DSA of 2.3. As such, this stretchable gel loaded with quinine was used to inhibit the growth of E. coli in lysogeny broth only when the gel was stretched. We believe the results of this study would be valuable for improving the quality of controlled molecular delivery and subsequent efficacy of molecular cargos.

Poly(ethylene glycol)-Mediated Collagen Gel Mechanics Regulates Cellular Phenotypes in a Microchanneled Matrix.

For the past few decades, efforts have been extensively made to reproduce tissue of interests for various uses including fundamental bioscience studies, clinical treatments, and even soft robotic systems. In these studies, cells are often cultured in micropores introduced in a provisional matrix despite that bulk rigidity may negatively affect cellular differentiation involved in tissue formation. To this end, we hypothesized that suspending cells within a soft fibrous matrix that is encapsulated within the microchannels of a provisional matrix would allow us to mediate effects of the matrix rigidity on cells and, in turn, to increase the cell differentiation level. We examined this hypothesis by filling microchannels interpenetrating alginate matrices with collagen gels of controlled elastic moduli (i.e., 125 to 1 Pa). Myoblasts used as a model predifferentiated cell were suspended within the collagen gels. The elastic modulus of the collagen gels was decreased through the addition of poly(ethylene glycol) during the gel preparation. Myoblasts loaded in the collagen gel exhibited a higher myogenic differentiation level than those adhered to the collagen-coated microchannel wall. Furthermore, the collagen gel softened by poly(ethylene glycol) further increased the volume of the multinucleated myofibers. The role of collagen gel softness on cell differentiation became more significant when the bulk elastic modulus of the alginate matrix was tuned to be close to that of muscle tissue (i.e., 11 kPa). We believe that the results of this study would be useful to understanding phenotypic activities of a wide array of cells involved in tissue development and regeneration.