Varying the Stiffness and Diffusivity of Rod-Shaped Microgels Independently through Their Molecular Building Blocks.

Microgels are water-swollen, crosslinked polymers that are widely used as colloidal building blocks in scaffold materials for tissue engineering and regenerative medicine. Microgels can be controlled in their stiffness, degree of swelling, and mesh size depending on their polymer architecture, crosslink density, and fabrication method-all of which influence their function and interaction with the environment. Currently, there is a lack of understanding of how the polymer composition influences the internal structure of soft microgels and how this morphology affects specific biomedical applications. In this report, we systematically vary the architecture and molar mass of polyethylene glycol-acrylate (PEG-Ac) precursors, as well as their concentration and combination, to gain insight in the different parameters that affect the internal structure of rod-shaped microgels. We characterize the mechanical properties and diffusivity, as well as the conversion of acrylate groups during photopolymerization, in both bulk hydrogels and microgels produced from the PEG-Ac precursors. Furthermore, we investigate cell-microgel interaction, and we observe improved cell spreading on microgels with more accessible RGD peptide and with a stiffness in a range of 20 kPa to 50 kPa lead to better cell growth.

Microgels as Platforms for Antibody-Mediated Cytokine Scavenging.

Therapeutic antibodies are the key treatment option for various cytokine-mediated diseases, such as rheumatoid arthritis, psoriasis, and inflammatory bowel disease. However, systemic injection of these antibodies can cause side effects and suppress the immune system. Moreover, clearance of therapeutic antibodies from the blood is limiting their efficacy. Here, water-swollen microgels are produced with a size of 25 µm using droplet-based microfluidics. The microgels are functionalized with TNFα antibodies to locally scavenge the pro-inflammatory cytokine TNFα. Homogeneous distribution of TNFα-antibodies is shown throughout the microgel network and demonstrates specific antibody-antigen binding using confocal microscopy and FLIM-FRET measurements. Due to the large internal accessibility of the microgel network, its capacity to bind TNFα is extremely high. At a TNFα concentration of 2.5 µg mL-1 , the microgels are able to scavenge 88% of the cytokine. Cell culture experiments reveal the therapeutic potential of these microgels by protecting HT29 colorectal adenocarcinoma cells from TNFα toxicity and resulting in a significant reduction of COX II and IL8 production of the cells. When the microgels are incubated with stimulated human macrophages, to mimic the in vivo situation of inflammatory bowel disease, the microgels scavenge almost all TNFα that is produced by the cells.

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.

Functionalized Microgel Rods Interlinked into Soft Macroporous Structures for 3D Cell Culture.

In this work, a two component microgel assembly using soft anisometric microgels that interlink to create a 3D macroporous construct for cell growth is reported. Reactive microgel rods with variable aspect ratio are produced via microfluidics in a continuous plug-flow on-chip gelation method by photoinitiated free-radical polymerization of star-polyethylene glycol-acrylate with glycidyl methacrylate or 2-aminoethyl methacrylate comonomers. The resulting complementary epoxy- and amine-functionalized microgels assemble and interlink with each other via a ring opening reaction, resulting in macroporous constructs with pores up to several hundreds of micrometers. The level of crosslinking depends on the functionalization degree of the microgels, which also affects the stiffness and cell adhesiveness of the microgels when modified with the cell-adhesive GRGDS-PC peptide. Therefore, 3D spreading and growth of cells inside the macroporous structure is influenced not only by the presence of macropores but also by the mechanical and biochemical properties of the individual microgels.

Translating Therapeutic Microgels into Clinical Applications.

Microgels are crosslinked, water-swollen networks with a 10 nm to 100 µm diameter and can be modified chemically or biologically to render them biocompatible for advanced clinical applications. Depending on their intended use, microgels require different mechanical and structural properties, which can be engineered on demand by altering the biochemical composition, crosslink density of the polymer network, and the fabrication method. Here, the fundamental aspects of microgel research and development, as well as their specific applications for theranostics and therapy in the clinic, are discussed. A detailed overview of microgel fabrication techniques with regards to their intended clinical application is presented, while focusing on how microgels can be employed as local drug delivery materials, scavengers, and contrast agents. Moreover, microgels can act as scaffolds for tissue engineering and regeneration application. Finally, an overview of microgels is given, which already made it into pre-clinical and clinical trials, while future challenges and chances are discussed. This review presents an instructive guideline for chemists, material scientists, and researchers in the biomedical field to introduce them to the fundamental physicochemical properties of microgels and guide them from fabrication methods via characterization techniques and functionalization of microgels toward specific applications in the clinic.