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Polyacrylamide (PAAm) hydrogels are widely adopted as 2D‐model soft substrates for investigating cell‐material interactions in a controlled in vitro environment. They offer facile synthesis, tunable physico‐chemical properties, diverse biofunctionalization routes, optical transparency, mouldability in a range of geometries and shapes, and compatibility with living cells. PAAm hydrogels can be engineered to reconstruct physiologically relevant biointerfaces, like cell‐matrix or cell–cell interfaces, featuring biochemical, mechanical, and topographical cues present in the extracellular environment. This Review provides a materials science perspective on PAAm material properties, fabrication, and modification strategies relevant to cell studies, highlighting their versatility and potential to address a wide range of biological questions. Current routes are presented to integrate cell‐instructive features, such as 2D patterns, 2.5D surface topographies, or mechanical stiffness gradients. Finally, the recent advances are emphasized toward dynamic PAAm hydrogels with on‐demand control over hydrogel properties as well as electrically conductive PAAm hydrogels for bioelectronics. Polyacrylamide (PAAm) hydrogels are versatile soft substrates commonly used in cell‐material studies. This Review explores their fabrication, physico‐chemical properties, and modification strategies to incorporate features like 2D patterns, surface topographies, and mechanical stiffness gradients. Additionally, recent advances in photoresponsive and electrically conductive PAAm hydrogels for bioelectronics are discussed.

Biofunctionalized polyacrylamide (PAAm) hydrogels are important 2D substrates for studying cell physics and mechanobiology. In this work, an arylmethylsulfone (MS) comonomer is developed that can be incorporated into PAAm gels under aqueous radical polymerization conditions. The resulting hydrogels show similar properties to unmodified PAAm gels, indicating that the comonomer is incorporated without affecting PAAm physical properties. The MS‐containing PAAm hydrogels allow efficient conjugation of thiol derivatized biomolecules and require very low comonomer content (2 mM, 0.18 mol% relative to AAm) and thiol incubation amounts (≥ 0.15 µg per gel) to achieve functional densities that elicit cell responses. Compared to carboxyl‐functionalized PAAm hydrogels, a 10‐fold lower comonomer concentration and a 10‐fold lower ligand feed concentration are sufficient to achieve comparable cell adhesion responses. The new comonomer opens up possibilities for efficient and straightforward biofunctionalization of PAAm hydrogels used in cell biophysical studies. A new methylsulfone comonomer is presented that can be integrated into polyacrylamide hydrogels for efficient biofunctionalization with thiol‐bearing ligands. Very low comonomer and ligand incubation amounts are required to achieve ligand densities that elicit cell responses, offering an efficient pathway to bioactive hydrogel surfaces for cell physics and mechanobiology studies.