Ludwig Boltzmann Institute for Traumatology, The Research Center in Cooperation with AUVA, Austrian Cluster for Tissue Regeneration, Vienna, Austria.
Medical University of Vienna, Department of Orthopedics and Trauma Surgery, Division of Trauma Surgery, Vienna, Austria.
Methods Mol Biol. 2023;2598:301-311. doi: 10.1007/978-1-0716-2839-3_21.
Removing cellular material from a tissue, a process called decellularization, reduces the risk of adverse host reactions, allows for efficient decontamination, and extends the shelf-life of the matrix. It facilitates the use of cartilage tissue as human-derived allograft, thus providing the field of cartilage regeneration with a biomaterial unmatched in its similarity to native cartilage in terms of structure, composition, and mechanical properties.The dense extracellular matrix of articular cartilage requires a particularly thorough process to achieve the removal of cells, cell debris, and reagents used in the process. In our studies (Nürnberger et al., EBioMedicine 64:103196, 2021; Schneider et al., Tissue Eng Part C Methods 22(12):1095-1107, 2016), we have successfully developed a protocol for achieving decellularization via physical, chemical, and enzymatic steps. Combining freeze-thaw cycles for devitalization, hydrochloric acid as decellularization agent and the enzymatic removal of glycosaminoglycans, results in an acellular scaffold that is fully biocompatible and promotes cellular attachment. The structure and sophisticated architecture of collagen type II is left intact.This chapter provides a comprehensive guide to the steps and reagents needed to decellularize articular cartilage. In addition to the standard decell-deGAG protocol, a fast option is given which is suitable for thin specimen. Histological evaluation is presented to illustrate treatment success.
从组织中去除细胞物质，即脱细胞过程，降低了宿主不良反应的风险，使有效的去污成为可能，并延长了基质的保质期。它促进了软骨组织作为人类来源的同种异体移植物的使用，从而为软骨再生领域提供了一种生物材料，在结构、组成和机械性能方面与天然软骨非常相似。关节软骨的密集细胞外基质需要特别彻底的过程才能去除细胞、细胞碎片和过程中使用的试剂。在我们的研究中（Nürnberger 等人，EBioMedicine 64:103196，2021 年；Schneider 等人，组织工程部分 C 方法 22(12):1095-1107，2016 年），我们已经成功开发了一种通过物理、化学和酶步骤实现脱细胞的方案。结合用于失活的冻融循环、盐酸作为脱细胞剂和糖胺聚糖的酶去除，产生完全生物相容并促进细胞附着的无细胞支架。胶原 II 型的结构和复杂架构得以保留。本章提供了一个全面的指南，介绍了脱细胞化关节软骨所需的步骤和试剂。除了标准的脱细胞去糖胺聚糖方案外，还提供了一种适用于薄片的快速选项。提供了组织学评估以说明处理成功。
