Decellularization and preservation of human skin: A platform for tissue engineering and reconstructive surgery

• Published in: Methods (San Diego, Calif.) Vol. 171; pp. 62 - 67
• Main Authors: Brouki Milan, Peiman, Pazouki, Abdolreza, Joghataei, Mohammad Taghi, Mozafari, Masoud, Amini, Naser, Kargozar, Saeid, Amoupour, Moein, Latifi, Noorahmad, Samadikuchaksaraei, Ali
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-01-2020
• Subjects: Acellular Dermis - trends; Animals; Cryopreservation; Decellularization; Dermal substitute; Extracellular matrix; Extracellular Matrix - chemistry; Extracellular Matrix - transplantation; Human skin; Humans; Reconstructive Surgical Procedures - trends; Skin - chemistry; Skin - cytology; Tissue Engineering - trends; Tissue Scaffolds - chemistry; Extracellular matrix; Human skin; Dermal substitute; Decellularization
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2019.07.005

•A highly reproducible method is needed for preparation of any regenerative modality.•An effective protocol for human skin decellularization is still to be developed.•We report a reproducible method for preparation of off-the-shelf decellularized human skin.
A matrix derived from natural tissue functions as a highly biocompatible and versatile scaffold for tissue engineering applications. It can act as a supportive construct that provides a niche for colonization by host cells. In this work, we describe a cost-effective, reliable and reproducible protocol for decellularization and preservation of human skin as a potential soft tissue replacement. The decellularized human skin is achieved using purely chemical agents without any enzymatic steps. The suitability of the proposed method for the preservation of the extracellular matrix (ECM) structure and its main components and integrity were evaluated using histological and immunohistochemical analysis. Cryopreservation and final sterility were conducted using programmable freeze-drying and gamma irradiation. The architecture, basement membrane and 3D structure of ECM can be successfully preserved after decellularization. Our protocol was found to be appropriate to maintain key proteins such as collagen type I, III, IV and laminin in the structure of final scaffold. This protocol offers a novel platform for the preparation of a dermal substitute for potential clinical applications.
Clinical application of naturally-based scaffolds for verity of health problems obliges development of a reproducible and effective technology that does not change structural and compositional material properties during scaffold preparation and preservation. Lack of an effective protocol for the production of biological products using decellularization method is still remaining. This effort is directing to solve this challenge in order to accomplish the off-the –shelf availability of decellularized dermal scaffold in market for clinical application.