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

## TL;DR

A new method using supercritical CO2 helps create better bioactive scaffolds for healing skin wounds.

## Contribution

A novel scCO2-assisted co-solvent system for decellularization that preserves bioactivity and mechanical properties.

## Key findings

- scCO2-derived ADM showed enhanced bioactive component retention and better mechanical properties.
- In vitro tests showed good biocompatibility with L929 cells and minimal cytotoxicity.
- In mice, the ADM accelerated wound healing and improved collagen and angiogenesis.

## Abstract

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.