ECM-Preserving Decellularization of Dermis via Subtle Cell Membrane Disruption Induced by Subcritical Dimethyl Ether.

Decellularized tissues are used as biomaterials for transplantation. Many decellularized tissues in clinical applications are prepared using surfactants; however, we have developed a new decellularization method that uses subcritical dimethyl ether (DME) instead of surfactants. Subcritical DME perfusion is usually used for lipid extraction; therefore, by perfusing tissues with subcritical DME, phospholipid cell membranes may be destroyed. DME vaporizes at room temperature and pressure, therefore, it is expected that it will not remain in the decellularized tissues and will not be toxic. In this study, subcritical DME was perfused into the porcine dermis, and the sample was subjected to DNA degradation to produce a subcritical DME-decellularized dermis. The subcritical DME-decellularized dermis showed good cell response in vitro and in vivo. In addition, we investigated the mechanism of the subcritical DME decellularization method and found that surfactants dissolve the entire cell and almost remove it; however, subcritical DME causes minor damage to the cell membrane and removes the cell nucleus through DNase treatment while leaving some of the cell membrane intact. These results suggest that subcritical DME-decellularized dermis is nontoxic and has the potential to develop highly functional decellularized tissues, such as extracellular vesicles, unlike decellularized dermis prepared with surfactants.