A Single‐Cell RNA Sequencing Guided Multienzymatic Hydrogel Design for Self‐Regenerative Repair in Diabetic Mandibular Defects

Abstract Conventional bone tissue engineering materials struggle to reinstate physiological bone remodeling in a diabetic context, primarily due to the compromised repolarization of proinflammatory macrophages to anti‐inflammatory macrophages. Here, leveraging single‐cell RNA sequencing (scRNA‐seq) technology, the pivotal role of nitric oxide (NO) and reactive oxygen species (ROS) is unveiled in impeding macrophage repolarization during physiological bone remodeling amidst diabetes. Guided by scRNA‐seq analysis, we engineer a multienzymatic bone tissue engineering hydrogel scaffold (MEBTHS) composed is engineered of methylpropenylated gelatin hydrogel integrated with ruthenium nanozymes, possessing both Ru 0 and Ru 4+ components. This design facilitates efficient NO elimination via Ru 0 while simultaneously exhibiting ROS scavenging properties through Ru 4+ . Consequently, MEBTHS orchestrates macrophage reprogramming by neutralizing ROS and reversing NO‐mediated mitochondrial metabolism, thereby rejuvenating bone marrow‐derived mesenchymal stem cells and endothelial cells within diabetic mandibular defects, producing newly formed bone with quality comparable to that of normal bone. The scRNA‐seq guided multienzymatic hydrogel design fosters the restoration of self‐regenerative repair, marking a significant advancement in bone tissue engineering.