Facile Biofabrication of Heterogeneous Multilayer Tubular Hydrogels by Fast Diffusion-Induced Gelation

Multilayer (ML) hydrogels are useful to achieve stepwise and heterogeneous control over the organization of biomedical materials and cells. There are numerous challenges in the development of fabrication approaches toward this, including the need for mild processing conditions that maintain the integrity of embedded compounds and the versatility in processing to introduce desired complexity. Here, we report a method to fabricate heterogeneous multilayered hydrogels based on diffusion-induced gelation. This technique uses the quick diffusion of ions and small molecules (i.e., photoinitiators) through gel-sol or gel-gel interfaces to produce hydrogel layers. Specifically, ionically (e.g., alginate-based) and covalently [e.g., gelatin methacryloyl (GelMA-based)] photocross-linked hydrogels are generated in converse directions from the same interface. The ML (e.g., seven layers) ionic hydrogels can be formed within seconds to minutes with thicknesses ranging from tens to hundreds of micrometers. The thicknesses of the covalent hydrogels are determined by the reaction time (or the molecule diffusion time). Multiwalled tubular structures (e.g., mimicking branched multiwalled vessels) are mainly investigated in this study based on a removable gel core, but this method can be generalized to other material patterns. The process is also demonstrated to support the encapsulation of viable cells and is compatible with a range of thermally reversible core materials (e.g., gelatin and Pluronic F127) and covalently cross-linked formulations (e.g., GelMA and methacrylated hyaluronic acid). This biofabrication process enhances our ability to fabricate a range of structures that are useful for biomedical applications.