Ambiguity in the Presentation of Decellularized Tissue Composition: The Need for Standardized Approaches

Abstract

Decellularization offers great potential to the field of tissue engineering, as this method gives rise to scaffold material with the native organ architecture by removing all cellular material and leaving much of the extracellular matrix (ECM) intact. However, many parameters may affect decellularization efficacy and ECM retention and, therefore, decellularization protocols need to be optimized for specific needs. This requires robust methods for comparison of decellularized tissue composition. Various representation methods are used in literature to express tissue composition (DNA, glycosaminoglycans, collagen, other ECM proteins, and growth factors). Here, we present and compare the various methods used and demonstrate that normalization to either dry or wet decellularized weight might be misleading and may overestimate true component retention. Moreover, the magnitude of the confounding effect is likely to be decellularization treatment dependent. As a result, we propose alternative comparison strategies: normalization to whole organ or to a unit of whole initial organ weight. We believe proper assessment of decellularized tissue composition is paramount for the successful comparison of different decellularization protocols and clinical translation.

Decellularized liver matrix as substrates for rescue of acute hepatocytes toxicity

“…Liver cirrhosis is accompanied by abnormal changes in ECM structure and composition (Klaas et al, ), remodeling in the ECM composition may alter cell signaling and facilitate liver regeneration for liver diseases (Klaas et al, ; Williams, Clouston, & Forbes, ). On the other hand, the decellularization procedure without using harsh detergents may result in structural integrity of ECM (Maghsoudlou et al, ); moreover, Triton X‐100, a mild detergent and typically used for liver decellularization (Badylak, Taylor, & Uygun, ), can only reduce cell component/DNA content without depleting any glycosaminoglycans (GAGs) during the washing process (Bruyneel & Carr, ). The effect could be attributed that GAGs and other components (collagen, proteoglycans) constitute the liver extracellular microenvironment, and also play an important role in hepatic functions and viability (Damania et al, ; Lee et al, ); furthermore, the wound ECM of GaIN‐damaged cells can be subsequently replaced by mature ECM and return to the preinjury state (Jonker, Dijkhuis, Boes, Hardonk, & Grond, ).…”

Applications of decellularized extracellular matrix in bone and cartilage tissue engineering

“…Decellularization refers to the process of treating a tissue with any combination of physical stress and chemical/enzymatic agents to remove cellular components, leaving behind only the noncellular ECM that can be used for therapeutic applications. The specific method of decellularization used depends on the tissue type; for instance, while cartilage tissue is able to undergo a relatively harsh treatment, lung tissue requires a more sensitive decellularization method to preserve its tissue composition . The resulting decellularized extracellular matrix (dECM) can then be processed further for different tissue engineering applications.…”

“…The specific method of decellularization used depends on the tissue type; for instance, while cartilage tissue is able to undergo a relatively harsh treatment, lung tissue requires a more sensitive decellularization method to preserve its tissue composition. 14 The resulting decellularized extracellular matrix (dECM) can then be processed further for different tissue engineering applications. These applications are summarized in Table 1.…”

Bovine Decellularized Amniotic Membrane: Extracellular Matrix as Scaffold for Mammalian Skin

“…ANOVA test showed that no significant differences were evidenced in the tested protocols. It has been reported that a lower concentration of <50 ng/mg in a membrane implies that the matrix can be considered as decellularized [32,56]. Therefore, the chemical and mechanical methods used were effective at eliminating the DNA content from the DM.…”