Push out test; Integration; Chondrogenesis; TGF-β3; Stem cells; Chondrocytes.
Vinardell, T; Thorpe, S D; Buckley, C T; Kelly, D J, Chondrogenesis and integration of mesenchymal stem cells within an in vitro cartilage defect repair model, Annals of Biomedical Engineering, 37, 12, 2009, 2556-2565
Annals of Biomedical Engineering 37 12
Integration of repair tissue is a key indicator of the long-term success of cell-based
therapies for cartilage repair. The objective of this study was to compare the in vitro
chondrogenic differentiation and integration of agarose hydrogels seeded with either
chondrocytes or bone marrow derived mesenchymal stem cells (MSCs) in defects
created in cartilage explants. Chondrocytes and MSCs were isolated from porcine
donors, suspended in 2% agarose and then injected into cylindrical defects within the
explants. These constructs were maintained in a chemically defined medium
supplemented with 10 ng/ml of TGF-β3. Cartilage integration was assessed by
histology and mechanical push-out tests. After 6 weeks in culture, chondrocyte seeded
constructs demonstrated a higher integration strength (64.4 ± 8.3 kPa) compared to
MSC seeded constructs (22.7 ± 5.9 kPa). GAG (1.27 ± 0.3 kPa vs 0.19 ± 0.03 kPa)
and collagen (0.31 ± 0.08 kPa vs 0.09 ± 0.01 kPa) accumulation in chondrocyte
seeded constructs was greater than that measured in the MSC seeded group. The
GAG, collagen and DNA content of both chondrocyte and MSC-seeded hydrogels
cultured in cartilage explants was significantly lower than control constructs cultured
in free swelling conditions. The results of this study suggest that the explant model
may constitute a more rigorous in vitro test to assess MSC therapies for cartilage
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