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

Abstract

Hydrogel precursors are liquid solutions that are prone to leaking after surgical placement. This problem was overcome by incorporating either decellularized cartilage (DCC) or devitalized cartilage (DVC) microparticles into traditional photocrosslinkable hydrogel precursors in an effort to achieve a paste-like hydrogel precursor. DCC and DVC were selected specifically for their potential to induce chondrogenesis of stem cells, given that materials that are chondroinductive on their own without growth factors are a revolutionary goal in orthopedic medicine. We hypothesized that DVC, lacking the additional chemical processing steps in DCC to remove cell content, would lead to a more chondroinductive hydrogel with rat bone marrow-derived mesenchymal stem cells. Hydrogels composed of methacrylated hyaluronic acid (MeHA) and either DCC or DVC microparticles were tested with and without exposure to transforming growth factor (TGF)-b 3 over a 6 week culture period, where swelling, mechanical analysis, and gene expression were observed. For collagen II, Sox-9, and aggrecan expression, MeHA precursors containing DVC consistently outperformed the DCC-containing groups, even when the DCC groups were exposed to TGF-b 3 . DVC consistently outperformed all TGF-b 3 -exposed groups in aggrecan and collagen II gene expression as well. In addition, when the same concentrations of MeHA with DCC or DVC microparticles were evaluated for yield stress, the yield stress with the DVC microparticles was 2.7 times greater. Furthermore, the only MeHA-containing group that exhibited shape retention was the group containing DVC microparticles. DVC appeared to be superior to DCC in both chondroinductivity and rheological performance of hydrogel precursors, and therefore DVC microparticles may hold translational potential for cartilage regeneration.

Cartilage extracellular matrix as a biomaterial for cartilage regeneration

“…While press‐fit scaffolds may be appropriate for regularly shaped cartilage defects, it may not be suitable for irregularly shaped defects. Our research group has introduced paste‐like cartilage matrix–based materials that are intended to fill irregularly shaped cartilage defects, where the paste can be crosslinked after placement to stay in place . Investigating and expanding the different forms of cartilage matrix–based products are needed to encompass various patient‐specific applications.…”

“…Decellularized cell‐derived matrices and devitalized cell‐derived matrices are derived from secreted ECM materials from cells in vitro that are subject to decellularization procedures or only physical methods, respectively. The feasibility of using cartilage matrix was determined in several initial studies that demonstrated its potential chondroinductivity . During the initial studies, similar challenges arose and resulted in parallel developmental themes in the fabrication of cartilage ECM scaffolds using both DCC and DVC.…”

Applications of decellularized extracellular matrix in bone and cartilage tissue engineering

“…Tissues that undergo only physical decellularization, specifically freeze‐thawing, are considered to be devitalized but not decellularized, as the cells have been lysed, but the cell debris and genetic material still remain within the processed tissue. These samples are most often processed into particles, during which the tissues go through a combination of freeze‐thawing and lyophilization, and are then ground into powder using a freezer/mill . Devitalized tissue particles have been shown to have higher quantities of ECM components, such as GAGs, than those that have been additionally treated with chemical/enzymatic decellularization methods .…”

Recent Advances in Photo-Crosslinkable Hydrogels for Biomedical Applications

“…Cells (chondrocytes or mesenchymal stem cells) are encapsulated in MeHA hydrogels functionalized with methacrylated chondroitin sulfate, devitalized cartilage microparticles or N-cadherin mimetic peptides (Figure 4C). These encapsulated cells demonstrate increased expression of cartilage markers and deposition of collagen type II compared with those encapsulated in pristine MeHA hydrogels [98,112,113]. In addition, MeHA/GelMA hybrid hydrogel has been used to engineer 3D in vitro fibrotic heart valve tissue models for understanding the process of fibrotic remodeling and calcification in heart valve diseases [107,114,115].…”