Chondrocytes and bone marrow-derived mesenchymal stem cells undergoing chondrogenesis in agarose hydrogels of solid and channelled architectures respond differentially to dynamic culture conditions

Abstract

The objective of this study was to investigate how a combination of different scaffold architectures and rotational culture would influence the functional properties of thick cartilaginous tissues engineered using either chondrocytes or bone marrow-derived mesenchymal stem cells (BM-MSCs). Expanded porcine chondrocytes and BM-MSCs were suspended in 2% agarose and cast in custom-designed moulds to produce either regular solid or channelled construct cylinders. The study consisted of three seperate experimental arms. First, chondrocyte and BM-MSC constructs were cultured in free swelling conditions for 9 weeks. Second, constructs were subjected to rotational culture for a period of 3 weeks. Finally, BM-MSC-seeded constructs were subjected to delayed rotational culture, in which constructs were first cultured for 3 weeks in free swelling conditions, followed by an additional 3 weeks in rotating culture conditions. Constructs were supplemented with TGFβ3 during the first 3 weeks of all experiments. The introduction of channels alone had little effect on the spatial patterns of tissue accumulation in either chondrocyte- or BM-MSC-seeded constructs. The two cell types responded differentially to rotational culture, resulting in the formation of a more homogeneous tissue in chondrocyte-seeded constructs, but significantly inhibiting chondrogenesis of BM-MSCs. This inhibition of chondrogenesis in response to dynamic culture conditions was not observed if BM-MSC-seeded constructs were first maintained in free swelling conditions for 3 weeks prior to rotation. The results of this study demonstrate that bioreactor culture conditions that are beneficial for chondrocyte-based cartilage tissue engineering may be suboptimal for BM-MSCs.