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

Area of Science:

• Biomaterials Engineering
• Tissue Engineering
• Regenerative Medicine

Background:

• Xenogeneic scaffolds from porcine skin are promising for human skin regeneration due to structural and biochemical similarities.
• Current decellularization methods often damage the extracellular matrix (ECM), compromising scaffold integrity and performance.

Purpose of the Study:

• To develop an optimized decellularization strategy for porcine skin to create a dermal scaffold (DEPOMA) with preserved ECM architecture and enhanced biological properties.
• To evaluate the efficacy of an ultrasound-assisted, low-detergent protocol in preserving ECM components and structural integrity.

Main Methods:

• Developed a decellularization protocol combining ultrasonication, hypertonic/hypotonic treatments, and reduced Triton X-100 exposure.
• Quantified cellular component removal (DNA) and retention of key ECM proteins (Laminin, Collagen IV, Elastin) using immunohistochemistry.
• Assessed scaffold ultrastructure and porosity via scanning electron microscopy (SEM) and image analysis.
• Evaluated mechanical properties using uniaxial tensile testing and biocompatibility through fibroblast metabolic activity assays.
• Tested scaffold integration in an ex vivo porcine wound model and compared performance against commercial alternatives.

Main Results:

• Achieved over 99% DNA removal while preserving significant amounts of key ECM proteins (76% Laminin, 66% Collagen IV, 889% Elastin).
• SEM and image analysis confirmed maintenance of native dermal ultrastructure with enhanced, uniform porosity.
• DEPOMA exhibited preserved mechanical properties comparable to native skin.
• Demonstrated a 3.4-fold increase in human fibroblast metabolic activity, indicating enhanced biocompatibility.
• Ex vivo wound model showed progressive host cell infiltration (147 µm depth at 21 days), indicating scaffold integration.
• Outperformed detergent-based scaffolds and commercial templates in cell viability and proliferation.

Conclusions:

• The ultrasound-assisted, low-detergent decellularization strategy effectively preserves ECM integrity, porosity, and mechanical properties of porcine skin scaffolds.
• DEPOMA exhibits superior biocompatibility and promotes host cell infiltration, making it a promising candidate for wound healing and regenerative medicine.
• This optimized decellularization approach offers a translationally relevant dermal scaffold for advanced therapeutic applications.