Decellularization of porcine dermis via supercritical CO₂-assisted co-solvent system for cutaneous wound healing

Efficient generation of bioactive and structurally preserved decellularized extracellular matrix (dECM) is critical for regenerative medicine applications. Conventional decellularization techniques, however, frequently utilize detergents such as sodium dodecyl sulfate and Triton X-100, which may compromise ECM integrity, diminish retention of bioactive components and impair mechanical properties. To address these limitations, we present a supercritical CO₂ (scCO₂)-assisted co-solvent strategy for efficient decellularization of porcine dermis, yielding a bioactive acellular dermal matrix (ADM) patch with high clinical potential in cutaneous wound healing. Porcine dermal sheets were first subjected to scCO₂-ethanol co-solvent treatment (28 MPa, 40°C, 6 h) followed by rapid depressurization. The solvent system was then switched to scCO₂-water co-solvent with concurrent ultrasonic washing. After subsequent depressurization, pure scCO₂ was introduced to achieve supercritical drying. Compared with detergent-based methods, scCO₂-derived ADM demonstrated enhanced retention of bioactive components and superior mechanical properties. In vitro experiments demonstrated excellent biocompatibility, promoting L929 cell proliferation and migration, with minimal cytotoxicity and low apoptosis rates. In a full-thickness skin defect model in mice, the scCO₂-derived ADM accelerated wound closure, enhanced collagen deposition and significantly improved angiogenesis and ECM remodeling. Overall, this scCO₂-based decellularization approach provides a robust, efficient and environmentally friendly platform for preparing high-performance dECM scaffolds, offering promising prospects for clinical applications in skin regeneration.