Integrating conductive electrodes into hydrogel-based microfluidic chips for real-time monitoring of cell response.

The conventional real-time screening in organs-on-chips is limited to optical tracking of pre-tagged cells and biological agents. This work introduces an efficient biofabrication protocol to integrate tunable hydrogel electrodes into 3D bioprinted-on-chips. We established our method of fabricating cell-laden hydrogel-based microfluidic chips through digital light processing-based 3D bioprinting. Our conductive ink includes poly-(3,4-ethylene-dioxythiophene)-polystyrene sulfonate (PEDOT: PSS) microparticles doped in polyethylene glycol diacrylate (PEGDA). We optimized the manufacturing process of PEDOT: PSS microparticles characterized our conductive ink for different 3D bioprinting parameters, geometries, and materials conditions. While the literature is limited to 0.5% w/v for PEDOT: PSS microparticle concentration, we increased their concentration to 5% w/v with superior biological responses. We measured the conductivity in the 3-15 m/m for a range of 0.5%-5% w/v microparticles, and we showed the effectiveness of 3D-printed electrodes for predicting cell responses when encapsulated in gelatin-methacryloyl (GelMA). Interestingly, a higher cellular activity was observed in the case of 5% w/v microparticles compared to 0.5% w/v microparticles. Electrochemical impedance spectroscopy measurements indicated significant differences in cell densities and spheroid sizes embedded in GelMA microtissues.

4D-Printable Photocrosslinkable Polyurethane-Based Inks for Tissue Scaffold and Actuator Applications.

4D printing recently emerges as an exciting evolution of conventional 3D printing, where a printed construct can quickly transform in response to a specific stimulus to switch between a temporary variable state and an original state. In this work, a photocrosslinkable polyethylene-glycol polyurethane ink is synthesized for light-assisted 4D printing of smart materials. The molecular weight distribution of the ink monomers is tunable by adjusting the copolymerization reaction time. Digital light processing (DLP) technique is used to program a differential swelling response in the printed constructs after humidity variation. Bioactive microparticles are embedded into the ink and the improvement of biocompatibility of the printed constructs is demonstrated for tissue engineering applications. Cell studies reveal above 90% viability in 1 week and ≈50% biodegradability after 4 weeks. Self-folding capillary scaffolds, dynamic grippers, and film actuators are made and activated in a humid environment. The approach offers a versatile platform for the fabrication of complex constructs. The ink can be used in tissue engineering and actuator applications, making the ink a promising avenue for future research.

Cell encapsulation in gelatin methacryloyl bioinks impairs microscale diffusion properties.

Light-assisted bioprinted gelatin methacryloyl (GelMA) constructs have been used for cell-laden microtissues and organoids. GelMA can be loaded by desired cells, which can regulate the biophysical properties of bioprinted constructs. We study how the degree of methacrylation (MA degree), GelMA mass concentration, and cell density change mass transport properties. We introduce a fluorescent-microscopy-based method of biotransport testing with improved sensitivity compared to the traditional particle tracking methods. The diffusion capacity of GelMA with a higher MA significantly decreased compared to a lower MA. Opposed to a steady range of linear elastic moduli, the diffusion coefficient in GelMA varied when cell densities ranged from 0 to 10 × 106 cells/ml. A comparative study of different cell sizes showed a higher diffusivity coefficient for the case of larger cells. The results of this study can help bioengineers and scientists to better control the biotransport characteristics in light-assisted bioprinted microtissues and organoids.

3D Bioprinted Hydrogel Microfluidic Devices for Parallel Drug Screening.

Conventional high-throughput screening (HTS) platforms suffer from the need for large cell volumes, high reagent consumption, significant assembly cost, and handling efforts. The assembly of three-dimensional (3D) bioprinted hydrogel-based microfluidic chips within platforms can address these problems. We present a continuous and seamless manufacturing approach to create a bioprinted microfluidic chips with a circular pattern scalable toward HTS platforms. Digital light processing 3D bioprinting is used to tune the local permeability of our chip, made of polyethylene glycol diacrylate and cell-laden gelatin methacryloyl, for creating predefined gradients of biochemical properties. We measured the flow-induced physical characteristics, the mass transport of drug agents, and the biological features of the proposed chip. We measured reactive oxygen species from the encapsulated cells through an integrated process and showed the capacity of the hydrogel-based chip for creating drug/agent gradients. This work introduces a chip design based on a hydrogel that can be changed and could be used for modern HTS platforms such as in vitro organoids.

Multi-material digital light processing bioprinting of hydrogel-based microfluidic chips.

Recent advancements in digital-light-processing (DLP)-based bioprinting and hydrogel engineering have enabled novel developments in organs-on-chips. In this work, we designed and developed a multi-material, DLP-based bioprinter for rapid, one-step prototyping of hydrogel-based microfluidic chips. A composite hydrogel bioink based on poly-ethylene-glycol-diacrylate (PEGDA) and gelatin methacryloyl (GelMA) was optimized through varying the bioprinting parameters such as light exposure time, bioink composition, and layer thickness. We showed a wide range of mechanical properties of the microfluidic chips for various ratios of PEGDA:GelMA. Microfluidic features of hydrogel-based chips were then tested using dynamic flow experiments. Human-derived tumor cells were encapsulated in 3D bioprinted structures to demonstrate their bioactivity and cell-friendly environment. Cell seeding experiments then validated the efficacy of the selected bioinks for vascularized micro-tissues. Our biofabrication approach offers a useful tool for the rapid integration of micro-tissue models into organs-on-chips and high-throughput drug screening platforms.

Multi-Organs-on-Chips for Testing Small-Molecule Drugs: Challenges and Perspectives.

Organ-on-a-chip technology has been used in testing small-molecule drugs for screening potential therapeutics and regulatory protocols. The technology is expected to boost the development of novel therapies and accelerate the discovery of drug combinations in the coming years. This has led to the development of multi-organ-on-a-chip (MOC) for recapitulating various organs involved in the drug-body interactions. In this review, we discuss the current MOCs used in screening small-molecule drugs and then focus on the dynamic process of drug absorption, distribution, metabolism, and excretion. We also address appropriate materials used for MOCs at low cost and scale-up capacity suitable for high-performance analysis of drugs and commercial high-throughput screening platforms.

3D Printing metamaterials towards tissue engineering.

The rapid growth and disruptive potentials of three-dimensional (3D) printing demand further research for addressing fundamental fabrication concepts and enabling engineers to realize the capabilities of 3D printing technologies. There is a trend to use these capabilities to develop materials that derive some of their properties via their structural organization rather than their intrinsic constituents, sometimes referred to as mechanical metamaterials. Such materials show qualitatively different mechanical behaviors despite using the same material composition, such as ultra-lightweight, super-elastic, and auxetic structures. In this work, we review current advancements in the design and fabrication of multi-scale advanced structures with properties heretofore unseen in well-established materials. We classify the fabrication methods as conventional methods, additive manufacturing techniques, and 4D printing. Following a comprehensive comparison of different fabrication methods, we suggest some guidelines on the selection of fabrication parameters to construct meta-biomaterials for tissue engineering. The parameters include multi-material capacity, fabrication resolution, prototyping speed, and biological compatibility.