Optimization of ultrasonication-assisted decellularization of porcine liver for biomimetic scaffold fabrication.

Decellularized extracellular matrix (dECM)-based hydrogels in previous research demonstrated as promising biomimetic scaffolds. However, conventional decellularization protocols are commonly based on harsh detergents that disrupt ECM integrity. While milder detergents such as sodium lauryl ether sulfate (SLES) and physical methods like ultrasonication have been proposed to reduce these negative effects, their combined application and the influence of ultrasonication parameters remain insufficiently explored. In this study, we systematically investigated the effect of ultrasonication-assisted decellularization on porcine liver tissue treated with SLES, with particular emphasis on ultrasonication exposure time. Decellularization efficiency and ECM preservation were evaluated through DNA, collagen, and glycosaminoglycan quantification, and structural integrity analysis. The impact of processing conditions on downstream applications was assessed by characterizing the gelation behavior, rheological properties, molecular weight distribution, and microstructure of the resulting dECM-based hydrogels. Biocompatibility was evaluated using HaCaT cells through metabolic assay and adhesion. Ultrasonication-assisted decellularization improved ECM preservation while maintaining effective decellularization, as all samples had < 50 ng of DNA per mg or dry tissue. Furthermore, it enabled the formation of hydrogels with enhanced physicochemical properties and increased cell viability compared to hydrogels obtained by detergent decellularization only. These findings demonstrate that controlled ultrasonication can reduce the need for harsh chemical treatments and improve scaffold preservation, highlighting its potential for optimizing decellularization protocols in tissue engineering applications.