Annealing High Aspect Ratio Microgels into Macroporous 3D Scaffolds Allows for Higher Porosities and Effective Cell Migration.

Growing millimeter-scaled functional tissue remains a major challenge in the field of tissue engineering. Therefore, microporous annealed particles (MAPs) are emerging as promising porous biomaterials that are formed by assembly of microgel building blocks. To further vary the pore size and increase overall MAP porosity of mechanically stable scaffolds, rod-shaped microgels with high aspect ratios up to 20 are chemically interlinked into highly porous scaffolds. Polyethylene glycol based microgels (width 10 µm, lengths up to 200 µm) are produced via in-mold polymerization and covalently interlinked into stable 3D scaffolds via epoxy-amine chemistry. For the first time, MAP porosities can be enhanced by increasing the microgel aspect ratio (mean pore sizes ranging from 39 to 82 µm, porosities from 65 to 90%). These porosities are significantly higher compared to constructs made from spherical or lower aspect ratio rod-shaped microgels. Rapid filling of the pores by either murine or primary human fibroblasts is ensured as cells migrate and grow extensively into these scaffolds. Overall, this study demonstrates that highly porous, stable macroporous hydrogels can be achieved with a very low partial volume of synthetic, high aspect ratio microgels, leading to large empty volumes available for cell ingrowth and cell-cell interactions.

Soft temperature-responsive microgels of complex shape in stop-flow lithography.

Stop-flow lithography (SFL) has emerged as a facile high-throughput fabrication method for µm-sized anisometric particles; yet, the fabrication of soft, anisometric microgels has not frequently been addressed in the literature. Furthermore, and to the best of the authors' knowledge, no soft, complex-shaped microgels with temperature-responsive behavior have been fabricated with this technology before. However, such microgels have tremendous potential as building blocks and actuating elements in rapidly developing fields, such as tissue engineering and additive manufacturing of soft polymeric building blocks, bio-hybrid materials, or soft micro-robotics. Given their great potential, we prove in this work that SFL is a viable method for the fabrication of soft, temperature-responsive, and complex-shaped microgels. The microgels, fabricated in this work, consist of poly(N-isopropylacrylamide) (pNIPAm), which is crosslinked with N,N'-methylenebis(acrylamide). The results confirm that the shape of the pNIPAm microgels is determined by the transparency mask, used in SFL. Furthermore, it is shown that, in order to realize stable microgels, a minimum threshold of crosslinker concentration of 2 wt% is required. Above this threshold, the stiffness of pNIPAm microgels can be deliberately altered by adjusting the concentration of the crosslinker. The fabricated pNIPAm microgels show the targeted temperature-responsive behavior. Within this context, temperature-dependent reversible swelling is confirmed, even for fractal-like geometries, such as micro snowflakes. Thus, these microgels provide the targeted unique combination of softness, shape complexity, and temperature responsiveness and increase the freedom of design for actuated building blocks.

Compartmentalized Jet Polymerization as a High-Resolution Process to Continuously Produce Anisometric Microgel Rods with Adjustable Size and Stiffness.

In the past decade, anisometric rod-shaped microgels have attracted growing interest in the materials-design and tissue-engineering communities. Rod-shaped microgels exhibit outstanding potential as versatile building blocks for 3D hydrogels, where they introduce macroscopic anisometry, porosity, or functionality for structural guidance in biomaterials. Various fabrication methods have been established to produce such shape-controlled elements. However, continuous high-throughput production of rod-shaped microgels with simultaneous control over stiffness, size, and aspect ratio still presents a major challenge. A novel microfluidic setup is presented for the continuous production of rod-shaped microgels from microfluidic plug flow and jets. This system overcomes the current limitations of established production methods for rod-shaped microgels. Here, an on-chip gelation setup enables fabrication of soft microgel rods with high aspect ratios, tunable stiffness, and diameters significantly smaller than the channel diameter. This is realized by exposing jets of a microgel precursor to a high intensity light source, operated at specific pulse sequences and frequencies to induce ultra-fast photopolymerization, while a change in flow rates or pulse duration enables variation of the aspect ratio. The microgels can assemble into 3D structures and function as support for cell culture and tissue engineering.

A catalyst-free, temperature controlled gelation system for in-mold fabrication of microgels.

Anisometric microgels are prepared via thermal crosslinking using an in-mold polymerization technique. Star-shaped poly(ethylene oxide-stat-propylene oxide) polymers, end-modified with amine and epoxy groups, form hydrogels, of which the mechanical properties and gelation rate can be adjusted by the temperature, duration of heating, and polymer concentration. Depending on the microgel stiffness, the rod-shaped microgels self-assemble into ordered or disordered structures.