Mesenchymal stromal cell-derived extracellular matrix influences gene expression of chondrocytes

Abstract

Decellularized extracellular matrix (ECM) has recently gained a lot of interest as an instructive biomaterial for regenerative medicine applications. In this study, the ability of adult human mesenchymal stem cell (hMSC)-derived ECM to rescue the phenotype of osteoarthritic (OA) chondrocytes and to further stimulate the differentiation of healthy (HL) chondrocytes was evaluated. ECMs were prepared by decellularizing hMSCs cultured in basic medium (BM) and chondrogenic medium (CM). The obtained ECM was then combined with a polymeric solution of Poly (ε-caprolactone) (PCL) dissolved in 1, 1, 1, 3, 3, 3-hexafluoro-2-propanol (HFIP) and electrospun meshes were fabricated. Electrospun ECM scaffolds were characterized using scanning electron microscopy (SEM) and picrosirius red staining was used to confirm the presence of collagen. OA and HL chondrocytes were cultured on scaffolds containing hMSC ECM in BM or CM and compared to PCL electrospun scaffolds without ECM. Metabolic activity and chondrogenic gene expression were assessed by Alamar blue assay and quantitative PCR (qPCR) analysis, respectively. The ECM presence resulted in a significant difference in chondrocyte metabolic activity compared to PCL scaffolds alone. HL chondrocytes cultured for 21 days in chondrogenic medium on electrospun scaffolds containing hMSC ECM from BM showed a significant increase in collagen II and aggrecan expression compared to hMSC ECM from CM and PCL scaffolds without ECM incorporation. No significant influence of hMSC ECM presence on the chondrogenic signature of OA chondrocytes was found. The influence of decellularized hMSC ECM on HL chondrocytes suggests that hMSC-derived ECM scaffolds are promising candidates for cartilage tissue engineering applications.

Cell-derived matrices for tissue engineering and regenerative medicine applications

“…Furthermore, this effect was TGF-β dependent, as the matrix alone was not sufficient to induce chondrogenesis 25 . In the reverse scenario, in vitro culture of chondrocytes on MSC-derived matrices had increased collagen type II and aggrecan expression in the presence of chondrogenic media 37 , but this improvement was only observed when the matrices were derived from MSCs cultured in a basic growth medium and not chondrogenic medium containing TGF-β3 and dexamethasone. Unfortunately, in vitro culture on either MSC-derived matrix did not rescue the phenotype of chondrocytes obtained from an osteoarthritic donor 37 .…”

Scaffolds Derived from ECM Produced by Chondrogenically Induced Human MSC Condensates Support Human MSC Chondrogenesis

“…In this work, 4-week-old sheets were the base materials, but it may be possible to use +MS engineered cartilage at an earlier time point to gain the potential advantage of having less GAG 27 while still benefiting from enhanced matrix production compared to that of −MS tissues. At the same time, scaffolds made by ECMs of chondrogenically induced hMSCs have been previously shown to be more chondroinductive than ECMs of undifferentiated hMSCs, 30 suggesting that some neocartilaginous ECM components of the base material may be desirable for development of cartilage tissues by the reseeded cells.…”

Polymers in Tissue Engineering

“…However, the addition of natural polymers such as collagen [57] or synthetic peptides nanofibers that coat hydrophobic polymers [48] were shown to improve the proliferative properties of the scaffold over bare polymer. Mesenchymal stem cells cultured in pro-chondrogenic media on electrospun nanofibers made with a blend of polycaprolactone and ECM derived from cultured MSCs expressed more pro-chondrogenic gene activity [58]. While the physical and mechanical architecture of scaffolds is critical, if the cell can't properly attach to the polymer used then the engineering is all for naught.…”