Microenvironment Preconditioning by Sequential OLA1 and Cytl1 Release Using a Double-Barrier Hydrogel for Osteoarthritis Therapy
Description
Osteoarthritis (OA) is a degenerative joint disease in which early mitochondrial dysfunction and cuproptosis blunt endogenous repair. Because these two processes demand intervention at different times, we asked whether a “microenvironment preconditioning” strategy could meet both, and built a double-barrier hydrogel (CFG@M/E) that releases OLA1 early and Cytl1 late. OLA1, a protein we identified by proteomic profiling of synovial fibroblast-derived exosomes, is delivered rapidly under ROS-enriched conditions to suppress cuproptosis, whereas Cytl1, identified by single-cell RNA sequencing, is released slowly and in a pH-dependent manner to drive matrix synthesis. In vitro, early OLA1 suppressed Elesclomol-induced cuproptosis, restored mitochondrial membrane potential, and lowered ROS, and the subsequent Cytl1 exposure then promoted anabolism (COL2A1/SOX9 up, MMP13 down). In a DMM mouse model, intra-articular CFG@M/E reduced OARSI scores, preserved cartilage, and restored locomotion without systemic toxicity. The double-barrier design thus matches the bioavailability of OLA1 and Cytl1 to the distinct pathological windows of OA—a more rational approach than co-delivering two drugs at once.