Fibronectin-Enriched Interface Using a Spheroid-Converged Cell Sheet for Effective Wound Healing

Cell sheets and spheroids are cell aggregates with excellent tissue-healing effects. However, their therapeutic outcomes are limited by low cell-loading efficacy and low extracellular matrix (ECM). Preconditioning cells with light illumination has been widely accepted to enhance reactive oxygen species (ROS)-mediated ECM expression and angiogenic factor secretion. However, there are difficulties in controlling the amount of ROS required to induce therapeutic cell signaling. Here, we develop a microstructure (MS) patch that can culture a unique human mesenchymal stem cell complex (hMSCcx), spheroid-attached cell sheets. The spheroid-converged cell sheet structure of hMSCcx shows high ROS tolerance compared to hMSC cell sheets owing to its high antioxidant capacity. The therapeutic angiogenic efficacy of hMSCcx is reinforced by regulating ROS levels without cytotoxicity using light (610 nm wavelength) illumination. The reinforced angiogenic efficacy of illuminated hMSCcx is based on the increased gap junctional interaction by enhanced fibronectin. hMSCcx engraftment is significantly improved in our novel MS patch by means of ROS tolerative structure of hMSCcx, leading to robust wound-healing outcomes in a mouse wound model. This study provides a new method to overcome the limitations of conventional cell sheets and spheroid therapy.