In Situ Formation of Microgel Array Via Patterned Electrospun Nanofibers Promotes 3D Cell Culture and Drug Testing in a Microphysiological System

A microphysiological system (MPS) is recently emerging as a promising alternative to the classical preclinical models, especially animal testing. A key factor for the construction of MPS is to provide a biomimetic three-dimensional (3D) cellular microenvironment. However, it still remains a challenge to introduce extracellular matrix (ECM)-like biomaterials such as hydrogels and nanofibers in a precise and spatiotemporal manner. Herein, we report a strategy to fabricate a MPS combining both electrospun nanofibers and hydrogels. The in situ formation of microsized hydrogel (microgel) array in MPS is realized by patterning electrospun poly(l-lactic acid) (PLLA)/Ca2+ nanofibers via a solvent-loaded agarose stamp and injecting an alginate solution to trigger the quick ionic cross-linking between alginate and Ca2+ released from patterned nanofibers. The one-on-one integration of electrospun nanofibers and microgels not only provides a 3D cellular microenvironment in designated regions in MPS but also improves the stability of these microenvironments under dynamic culture. In addition, due to the biocompatible properties of an ionic cross-linking reaction, patterned cell array can be achieved simultaneously during the microgel formation process. A breast cancer model is then built in MPS by coculturing human breast cancer cells and human fibroblasts in microgel array, and its application in drug (cisplatin) testing is evaluated. Our data prove that MPS-MA offers a more precise platform for drug testing to evaluate the drug concentration, duration time, cancer microenvironment, etc, mainly due to its successful construction of the biomimetic 3D cellular microenvironment.