Abstract
Engineered extracellular matrices (ECMs) that replicate complex in-vivo features have shown great potential in tissue engineering. Biocompatible hydrogel microstructures have been widely used to replace these native ECMs for physiologically relevant research. However, accurate reproduction of the 3D hierarchical and nonuniform mechanical stiffness inside one integrated microstructure to mimic the complex mechanical properties of native ECMs presents a major challenge. Here, by using digital holographic microscopy (DHM)-based stiffness imaging feedback, we propose a novel closed-loop control algorithm to achieve high-accuracy control of mechanical properties for hydrogel microstructures that recapitulate the physiological properties of native ECMs with high fidelity. During photoprinting, the photocuring area of the hydrogel is divided into microscale grid areas to locally control the photocuring process. With the assistance of a motorized microfluidic channel, the curing thickness is controlled with layer-by-layer stacking. The DHM-based stiffness imaging feedback allows accurate adjustment of the photocuring degree in every grid area to change the crosslinking network density of the hydrogel, thus enabling large-span and high-resolution modulation of mechanical properties. Finally, the gelatin methacrylate was used as a typical biomaterial to construct the high-fidelity biomimetic ECMs. The Young's modulus could be flexibly modulated in the 10 kPa to 50 kPa range. Additionally, the modulus gradient was accurately controlled to within 2.9 kPa. By engineering ECM with locally different mechanical properties, cell spreading along the stiff areas was observed successfully. We believe that this method can regenerate complex biomimetic ECMs that closely recapitulate in-vivo mechanical properties for further applications in tissue engineering and biomedical research.