Rapid and Controllable Multilayer Cell Sheet Assembly via Biodegradable Nanochannel Membranes

The ability to precisely arrange and control the assembly of diverse cell types into intricate three-dimensional (3D) structures remains a critical challenge in tissue engineering. Herein, we describe a versatile and programmable 3D cell sheet assembly technology by developing biodegradable nanochannel (BNC) membrane to fulfill this unmet need. This membrane, hierarchically assembled from two-dimensional nanomaterial aggregates, exhibits both exceptional fluid permeability and rapid biodegradation under physiological conditions. The unique properties of the BNC membrane enable precise spatial and temporal control over cell assembly, facilitating the creation of complex 3D cellular architectures. The BNC membrane was integrated with a programmable negative-pressure-based cell assembly strategy to form single and multi-cellular 3D sheets in a highly controllable manner. To demonstrate the feasibility and translatability of this technology in the field of tissue engineering, we devised approaches to screen stem cell-derived therapeutics with "core-shell" macrophage-fibroblast multi-cellular patterns and treat murine diabetic skin wounds via scaffold-free 3D adipose-derived mesenchymal stem cell (ADMSC) sheets. In summary, our results demonstrate that the BNC membrane-based 3D cell sheet assembly approach significantly advances current tissue engineering capabilities, offering substantial potential for both regenerative medicine applications and the development of physiologically relevant disease models.