Patterned ELR-Gelatin Hydrogels Enable Rapid Endothelial Monolayer Formation via Bioactive Matrix Chemistry and Surface Topography.

The endothelialization of organ-on-chip platforms and vascular implants is often limited by slow cell attachment and unstable monolayer formation. This work presents a scalable workflow that imprints micro- and nano-gratings into elastin-like recombinamer (ELR)-based hydrogels, enabling rapid endothelial cell capture and accelerating monolayer formation within 14 days. Three gelatin-ELR formulations are engineered, with 1H-NMR confirming incorporation of sequences designed to modulate bioactivity (ELR1: inert, ELR2: uPA-responsive, ELR3: RGD-adhesive). ELR incorporation generates fibrillar microstructures and enhances mechanical performance, yielding elastic-dominant networks suitable for high-fidelity pattern transfer. Using this library, the combined effects of ELR bioactivity and groove geometry on human iPSC-derived endothelial cells are evaluated. In a 15-min attachment assay, patterned ELR composites markedly improve cell retention compared to gelatin, with ELR2 on ∼350 and ∼4 µm grooves performing best, consistent with controlled, cell-mediated interfacial remodeling. This early advantage persists, as ELR2 and ELR3 hydrogels support rapid alignment and reach confluence by day 14, whereas gelatin remains subconfluent. By combining enhanced early capture with protease-regulated remodeling, ELR2 identifies a favorable design window. These results establish a materials design framework linking programmable ELR chemistry with surface topography to engineer endothelial interfaces, providing a versatile platform for vascular biomaterials and microphysiological systems.