Physical decellularization of fish skin utilizing electrical fields

Decellularized tissues have attracted considerable attention in tissue engineering and regenerative medicine due to their diverse sources and excellent biocompatibility. However, current decellularization techniques often compromise the integrity of the extracellular matrix, leaving harmful chemical residues or inadequately removing immunogenic cellular components. Consequently, the biocompatibility and clinical efficacy of decellularized tissues are undermined. To address these issues, a novel decellularization technique employing an electrical field has been proposed. In the resulting decellularized tissue, the residual DNA concentration was measured at 27.44 ± 7.27 ng/mg, satisfying the evaluation criteria (<50 ng/mg) in a significantly shorter process compared to chemical/enzymatic decellularization (∼2 h vs. ∼15 h) and without the use of toxic reagents. The microstructure was better preserved in electrically decellularized skin than in chemically/enzymatically decellularized skin, maintaining an interconnected porous microstructure that facilitates neo-tissue ingrowth. Cytotoxicity assessments confirmed the non-cytotoxic nature of electrically decellularized fish skin, with a cell survival rate (115.84 ± 10.78%) higher than that of chemically/enzymatically decellularized and native skins (105.57 ± 3.43% and 118.69 ± 6.53%, respectively). The hemolysis rate of decellularized skin using electrical techniques was the lowest (0.35 ± 0.26%) compared to that of chemically/enzymatically decellularized and native skins (1.089 ± 0.03% and 1.11 ± 0.06%), indicating non-hemolytic material. Furthermore, subcutaneous implantation demonstrated that electrically decellularized skin did not elicit severe immune responses and exhibited faster tissue integration than chemically/enzymatically decellularized skin. Therefore, electrical decellularization effectively removes cellular components while preserving the natural architecture without the use of harmful reagents, establishing a secure foundation for extensive applications in tissue engineering and regenerative medicine using decellularized tissues in the future.