Ultrasound-boosted low-detergent decellularization yields biocompatible skin scaffolds with native ECM integrity.

Xenogeneic scaffolds derived from porcine skin offer a promising alternative due to their structural and biochemical similarities to human skin. However, current decellularization strategies compromise extracellular matrix (ECM) integrity, porosity, or mechanical performance, limiting applicability. Here, we developed a decellularized porcine matrix (DEPOMA) scaffold using an ultrasound-assisted low-detergent strategy designed to achieve effective cellular removal while preserving ECM architecture. Specifically, we focused on combining ultrasonication, hypertonic/hypotonic treatments, and reduced Triton X-100 exposure. Our protocol effectively removed cellular components with over 99% deoxyribonucleic acid (DNA) removal, while preserving key basement membrane and dermal proteins, as seen by quantitative immunohistochemistry (IHC) demonstrating 76% Laminin, approximately 66% Collagen IV, and 889% Elastin retention relative to native tissue. Scanning electron microscopy (SEM) demostrated that DEPOMA maintained native dermal ultrastructure with enhanced and uniformly distributed porosity, quantified using DIGIMIZER image analysis. Uniaxial tensile testing on DEPOMA demonstrated preserved mechanical properties comparable to native skin. The DEPOMA scaffold demonstrated markedly enhanced biocompatibility, supporting a 3.4-fold increase in primary human fibroblast metabolic activity compared to controls. In a porcine full-thickness ex vivo wound model, DEPOMA showed progressive host-derived cellular infiltration reaching a penetration depth of 147 µm after 21 days, consistent with active scaffold integration and remodeling. When benchmarked against a detergent-based decellularized scaffold and a commercial dermal regeneration template, DEPOMA exhibited significantly improved cell viability and proliferative capacity. Collectively, these findings demonstrate that ultrasound-assisted low-detergent decellularization enables superior ECM preservation, structural integrity, and biological performance, supporting DEPOMA as a translationally optimized dermal scaffold for wound healing and regenerative medicine applications.