Decellularized Cartilage May Be a Chondroinductive Material for Osteochondral Tissue Engineering

Abstract

Extracellular matrix (ECM)-based materials are attractive for regenerative medicine in their ability to potentially aid in stem cell recruitment, infiltration, and differentiation without added biological factors. In musculoskeletal tissue engineering, demineralized bone matrix is widely used, but recently cartilage matrix has been attracting attention as a potentially chondroinductive material. The aim of this study was thus to establish a chemical decellularization method for use with articular cartilage to quantify removal of cells and analyze the cartilage biochemical content at various stages during the decellularization process, which included a physically devitalization step. To study the cellular response to the cartilage matrix, rat bone marrow-derived mesenchymal stem cells (rBMSCs) were cultured in cell pellets containing cells only (control), chondrogenic differentiation medium (TGF-β), chemically decellularized cartilage particles (DCC), or physically devitalized cartilage particles (DVC). The chemical decellularization process removed the vast majority of DNA and about half of the glycosaminoglycans (GAG) within the matrix, but had no significant effect on the amount of hydroxyproline. Most notably, the DCC group significantly outperformed TGF-β in chondroinduction of rBMSCs, with collagen II gene expression an order of magnitude or more higher. While DVC did not exhibit a chondrogenic response to the extent that DCC did, DVC had a greater down regulation of collagen I, collagen X and Runx2. A new protocol has been introduced for cartilage devitalization and decellularization in the current study, with evidence of chondroinductivity. Such bioactivity along with providing the ‘raw material’ building blocks of regenerating cartilage may suggest a promising role for DCC in biomaterials that rely on recruiting endogenous cell recruitment and differentiation for cartilage regeneration.

Chondroinduction from Naturally Derived Cartilage Matrix: A Comparison Between Devitalized and Decellularized Cartilage Encapsulated in Hydrogel Pastes

“…1A-C). In prior work, it was established that no significant reduction in biochemical content was observed between native cartilage and cartilage that was cryoground, 17 so the prior mentioned reductions are in reference to DVC powder.…”

“…17 In the pellet culture study, we observed that rat bone marrow stem cells (rBMSCs) exposed to DCC outperformed those cells exposed to DVC or transforming growth factorb 3 (TGF-b 3 ) in chondroinductivity. 17 However, gene expression was only observed over a period of 7 days and was only monitored for cells in pellet culture and not within a 3D scaffold.…”

“…We and other groups have already established that cartilage matrix has chondroinductive potential, 7,[13][14][15][16][17] and we recently were the first to compare the chondroinductive potential of two different types of cartilage matrix in pellet culture: devitalized cartilage (DVC), where the matrix was exposed to a freeze/thaw process to devitalize the living chondrocytes within the matrix, and decellularized cartilage (DCC), in which the cells were not only devitalized but also removed from the matrix entirely. 17 In the pellet culture study, we observed that rat bone marrow stem cells (rBMSCs) exposed to DCC outperformed those cells exposed to DVC or transforming growth factorb 3 (TGF-b 3 ) in chondroinductivity.…”

“…Our group was the first to explore the short-term chondrogenic potential of DCC versus DVC. 17 However, for the first time in this study, not only were DVC and DCC compared for their longterm chondrogenic potential, but they were also evaluated for their ability to exhibit a yield stress in hydrogel precursor solutions. In this study, chondrogenesis was induced through incorporating native cartilage ECM into the MeHA/cartilage matrix gel networks, which furthermore resulted in the pastelike behavior and yield stress that was observed before crosslinking.…”

Decellularized Bovine Articular Cartilage Matrix Reinforced by Carboxylated-SWCNT for Tissue Engineering Application

“…The nanotubes Can affect the cells and the cell morphology alter through the nanostructured surface. thus CNT-based surfaces provide a very good scaffold that can be used to stimulate the formation of cartilage tissue (Sutherland et al 2015 andTrzeciak et al 2016). We tried to keep natural structure of decellularized cartilage intact and put it close to nanotubes in such an environment.…”