Decellularization of Articular Cartilage: A Hydrochloric Acid-Based Strategy

Abstract

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.

Keywords: Cartilage; Decellularization; Decontamination; Glycosaminoglycans; Hydrochloric acid.