Hydration‐Mediated Negative Stress Hydrogel for 3D Cell Infiltration

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Abstract Biomechanical cell‐matrix interactions rely on both actin polymerization‐driven pushing and actomyosin contractility, yet synthetic scaffolds capable of recapitulating such ubiquitous mechanics remain unexplored. Herein, a frozen hydrogel (F‐gel) with reversible negative stress arising from a stratified collagen network is developed. The F‐gel exhibits a tunable force response, shifting from contraction (−27.77 kPa) at low collagen contents to resistance (184.61 kPa) once the fraction exceeds 2.0 wt.%. Molecular dynamic simulations and experiments reveal that this negative stress originates from the collapse of the stratified 3D network due to dynamic hydration crosslinks and polymer chain mobility. Leveraging orthogonal freezing to stratify the polymer network, this strategy is broadly applicable across diverse hydrogel systems with programmable regulation of negative and positive force response via solvent exchange. This unique mechanical behavior of F‐gel provides mechanical cues for highly efficient 3D cell infiltration. It is demonstrated that the biomimicry traction of F‐gel is promising in enhancing 3D cell spreading, orientation, migration, and infiltration, reaching 336.30 µm—over 20 fold deeper with 76 fold more cells than controls. These findings implicate an unrecognized paradigm for understanding the mechano‐transduction for tissue engineering and regenerative medicine.

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