Hydrogels with stiffness-degradation spatial patterns control anisotropic 3D cell response.

In nature, tissues are patterned, but most biomaterials used in human applications are not. Patterned biomaterials offer the opportunity to mimic spatially segregating biophysical and biochemical properties found in nature. Engineering such properties allows to study cell-matrix interactions in anisotropic matrices in great detail. Here, we developed alginate-based hydrogels with patterns in stiffness and degradation, composed of distinct areas of soft non-degradable (Soft-NoDeg) and stiff degradable (Stiff-Deg) material properties. The hydrogels exhibit emerging patterns in stiffness and degradability over time, taking advantage of dual crosslinking: Diels-Alder covalent crosslinking (norbornene-tetrazine, non degradable) and UV-mediated peptide crosslinking (matrix metalloprotease sensitive peptide, enzymatically degradable). The materials were mechanically characterized using rheology for single-phase and surface micro-indentation for patterned materials. 3D encapsulated mouse embryonic fibroblasts (MEFs) allowed to characterize the anisotropic cell-matrix interaction in terms of cell morphology by employing a novel image-based quantification tool. Live/dead staining showed no differences in cell viability but distinct patterns in proliferation, with higher cell number in Stiff-Deg materials at day 14. Patterns of projected cell area became visible already at day 1, with larger values in Soft-NoDeg materials. This was inverted at day 14, when larger projected cell areas were identified in Stiff-Deg. This shift was accompanied by a significant decrease in cell circularity in Stiff-Deg. The control of anisotropic cell morphology by the material patterns was also confirmed by a significant increase in filopodia number and length in Stiff-Deg materials. The novel image-based quantification tool was useful to spatially visualize and quantify the anisotropic cell response in 3D hydrogels with stiffness-degradation spatial patterns. Our results show that patterning of stiffness and degradability allows to control cell anisotropic response in 3D and can be quantified by image-based strategies. This allows a deeper understanding of cell-matrix interactions in a multicomponent material.

Implementing Zn2+ ion and pH-value control into artificial mussel glue proteins by abstracting a His-rich domain from preCollagen.

A His-rich domain of preCollagen-D found in byssal threads is derivatized with Cys and Dopa flanks to allow for mussel-inspired polymerization. Artificial mussel glue proteins are accessed that combine cysteinyldopa for adhesion with sequences for pH or Zn2+ induced ß-sheet formation. The artificial constructs show strong adsorption to Al2O3, the resulting coatings tolerate hypersaline conditions and cohesion is improved by activating the ß-sheet formation, that enhances E-modulus up to 60%.

Molding and Encoding Carbon Nitride-Containing Edible Oil Liquid Objects via Interfacial Toughening in Waterborne Systems.

Charge interaction-driven jamming of nanoparticle monolayers at the oil-water interface can be employed as a method to mold liquids into tailored stable 3D liquid objects. Here, 3D liquid objects are fabricated via a combination of biocompatible aqueous poly(vinyl sulfonic acid, sodium salt) solution and a colloidal dispersion of highly fluorescent organo-modified graphitic carbon nitride (g-C3N4) in edible sunflower oil. The as-formed liquid object shows stability in a broad pH range, as well as flexible pathways for efficient exchange of molecules at the liquid-liquid interphase, which allows for photodegradation of rhodamine B at the interface via visible light irradiation that also enables an encoding concept. The g-C3N4-based liquid objects point toward various applications, for example, all-liquid biphasic photocatalysis, artificial compartmentalized systems, liquid-liquid printing, or bioprinting.

Squid Suckerin Biomimetic Peptides Form Amyloid-like Crystals with Robust Mechanical Properties.

We present the self-assembly of fibers formed from a peptide sequence (A1H1) derived from suckerin proteins of squid sucker ring teeth (SRT). SRT are protein-only biopolymers with an unconventional set of physicochemical and mechanical properties including high elastic modulus coupled with thermoplastic behavior. We have identified a conserved peptide building block from suckerins that possess the ability to assemble into materials with similar mechanical properties as the native SRT. A1H1 displays amphiphilic characteristics and self-assembles from the bottom-up into mm-scale fibers initiated by the addition of a polar aprotic solvent. A1H1 fibers are thermally resistant up to 239 °C, coupled with an elastic modulus of ∼7.7 GPa, which can be explained by the tight packing of ß-sheet-enriched crystalline building blocks as identified by wide-angle X-ray scattering (WAXS), with intersheet and interstrand distances of 5.37 and 4.38 Å, respectively. A compact packing of the peptides at their Ala-rich terminals within the fibers was confirmed from molecular dynamics simulations, and we propose a hierarchical model of fiber assembly of the mature peptide fiber.

Bio-Inspired Mechanotactic Hybrids for Orchestrating Traction-Mediated Epithelial Migration.

A platform of mechanotactic hybrids is established by projecting lateral gradients of apparent interfacial stiffness onto the planar surface of a compliant hydrogel layer using an underlying rigid substrate with microstructures inherited from 3D printed molds. Using this platform, the mechanistic coupling of epithelial migration with the stiffness of the extracellular matrix (ECM) is found to be independent of the interfacial compositional and topographical cues.