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.

中文翻译：

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人体皮肤的脱细胞和保存：组织工程和重建手术的平台

源自天然组织的基质用作组织工程应用的高度生物相容性和多功能支架。它可以作为一种支持性结构，为宿主细胞的定殖提供一个利基。在这项工作中，我们描述了一种具有成本效益、可靠和可重复的协议，用于人类皮肤的脱细胞和保存，作为潜在的软组织替代品。脱细胞人体皮肤是使用纯化学试剂实现的，无需任何酶促步骤。使用组织学和免疫组织化学分析评估了所提出的保存细胞外基质 (ECM) 结构及其主要成分和完整性的方法的适用性。使用可编程冷冻干燥和伽马辐射进行冷冻保存和最终灭菌。架构，ECM 的基底膜和 3D 结构可以在脱细胞后成功保存。我们的协议被发现适用于在最终支架的结构中保持关键蛋白质，如 I、III、IV 型胶原蛋白和层粘连蛋白。该协议为制备潜在临床应用的皮肤替代品提供了一个新平台。