3D-Printed Hydrogels as Photothermal Actuators.

Thermoresponsive hydrogels were 3D-printed with embedded gold nanorods (GNRs), which enable shape change through photothermal heating. GNRs were functionalized with bovine serum albumin and mixed with a photosensitizer and poly(N-isopropylacrylamide) (PNIPAAm) macromer, forming an ink for 3D printing by direct ink writing. A macromer-based approach was chosen to provide good microstructural homogeneity and optical transparency of the unloaded hydrogel in its swollen state. The ink was printed into an acetylated gelatin hydrogel support matrix to prevent the spreading of the low-viscosity ink and provide mechanical stability during printing and concurrent photocrosslinking. Acetylated gelatin hydrogel was introduced because it allows for melting and removal of the support structure below the transition temperature of the crosslinked PNIPAAm structure. Convective and photothermal heating were compared, which both triggered the phase transition of PNIPAAm and induced reversible shrinkage of the hydrogel-GNR composite for a range of GNR loadings. During reswelling after photothermal heating, some structures formed an internally buckled state, where minor mechanical agitation recovered the unbuckled structure. The BSA-GNRs did not leach out of the structure during multiple cycles of shrinkage and reswelling. This work demonstrates the promise of 3D-printed, photoresponsive structures as hydrogel actuators.

Additive Soft Matter Design by UV-Induced Polymer Hydrogel Inter-Crosslinking.

In recent years, stimuli-responsive hydrogels have gained tremendous interest in designing complex smart 4D materials for applications ranging from biomedicine to soft electronics that can change their properties on demand over time. However, at present, a hydrogel's response is often induced by merely a single stimulus, restricting its broader applicability. The controlled hierarchical assembly of various hydrogel building blocks, each with a tailored set of mechanical and physicochemical properties as well as programmed stimulus response, may potentially enable the design and fabrication of multi-responsive polymer parts that process complex operations, like signal routing dependent on different stimuli. Since inter-connection stability of such building blocks directly accompanies the transmission of information across building blocks and is as important as the building property itself to create complex 4D materials, we provide a study on the utility of an inter-crosslinking mechanism based on UV-induced 2,3-dimethylmaleimide (DMMI) dimerization to inter-connect acrylamide-based and N-isopropylacrylamide-based millimeter-sized cubic building blocks, respectively. The resulting dual-crosslinked assemblies are freestanding and stable against contraction-expansion cycles in solution. In addition, the approach is also applicable for connecting microfluidically fabricated, micrometer-sized hydrogel spheres, with the resulting assemblies being processable and mechanical stable, likewise resisting contraction-expansion in different solvents, for instance.

Flexible Materials for High-Resolution 3D Printing of Microfluidic Devices with Integrated Droplet Size Regulation.

We develop resins for high-resolution additive manufacturing of flexible micromaterials via projection microstereolithography (PµSL) screening formulations made from monomer 2-phenoxyethyl acrylate, the cross-linkers Ebecryl 8413, tri(propyleneglycol) diacrylate or 1,3,5-triallyl-1,3,5-triazine-2,4,6(1H,3H,5H)-trione, the photoabsorber Sudan 1, and the photoinitiator diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide. PµSL-printed polymer micromaterials made from this resin library are characterized regarding achievable layer thickness depending on UV exposure energy, and for mechanical as well as optical properties. The best-candidate resin from this screening approach allows for 3D-printing transparent microchannels with a minimum cross section of approximately 35 × 46 µm2, which exhibit proper solvent resistance against water, isopropanol, ethanol, n-hexane, and HFE-7500. The mechanical properties are predestined for 3D-printing microfluidic devices with integrated functional units that require high material flexibility. Exemplarily, we design flexible microchannels for on-demand regulation of microdroplet sizes in microemulsion formation. Our two outlines of integrated droplet regulators operate by injecting defined volumes of air, which deform the droplet-forming microchannel cross-junction, and change the droplet size therein. With this study, we expand the library of functional resins for PµSL printing toward flexible materials with micrometer resolution and provide the basis for further exploration of these materials, e.g., as microstructured cell-culturing substrates with defined mechanics.

Cell-free protein synthesis and in situ immobilization of deGFP-MatB in polymer microgels for malonate-to-malonyl CoA conversion.

In the present work, microgels were utilized as a cell-free reaction environment to produce a functional malonyl-CoA synthetase (deGFP-MatB) under geometry-controlled transcription and translation. Our approach combines the straight-forward optimization of overall protein yield of an E. coli-based cell-free protein synthesis (CFPS) system based on concentration screening of magnesium and potassium glutamate, DNA as well as polyethylene glycol (PEG), and its innovative usage in microgel-based production of a key enzyme of the polyketide synthesis pathway. After partial modification of the carboxyl groups of hyaluronic acid (HA) with 5'-methylfuran groups via 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methyl-morpholinium chloride (DMTMM)-activation, these were further functionalized with dibenzocyclooctyne (DBCO) and nitrilotriacetic acid (NTA) groups by bio-orthogonal [4+2] Diels-Alder cycloaddition to yield a bifunctional macromer. After coupling the DBCO groups with azide-functionalized DNA, containing the genetic information for deGFP-MatB, via strain-promoted azide-alkyne cycloaddition (SPAAC), the DNA-/NTA-functionalized HA macromer was utilized as base material together with maleimide-functionalized PEG (PEG-mal2) as the crosslinker to form bifunctional microgels utilizing water-in-oil (W/O) microemulsions. As-formed microgels were incubated with nickel sulfate to activate the NTA groups and provide binding sites for deGFP-MatB, which contained six histidine residues (His-tag) for that purpose. The optimized CFPS mixture was loaded into the microgels to initiate the formation of deGFP-MatB, which was detected by a clear increase in fluorescence exclusively inside the microgel volume. Functionality of both, the bound and the decoupled enzyme was proven by reaction with malonate to yield malonyl CoA, as confirmed by a colorimetric assay.