A novel recirculating flow-perfusion bioreactor for periosteal chondrogenesis

Previous research indicated that engineered cartilage was soft and fragile due to less extracellular matrix than native articular cartilage. Consequently, the focus of this study was mostly confined to application in vitro function. In order to generate 3D engineered cartilage resembling native articular cartilage, we developed a recirculating flow-perfusion bioreactor to simulate the motion of a native diarthrodial joint by offering shear stress and hydrodynamic pressure simultaneously.

The bioreactor we developed offers steady oscillating laminar flow (maximum shear stress of 250 dyne/cm(2)) and hydrodynamic pressure (increased from 0 to 15 psi) simultaneously. The periosteal explants were harvested from the proximal medial tibiae of rabbits and fixed onto PCL scaffold with four corner sutures and cambium layer facing upward, then these periosteal composites (periosteum/ PCL) were placed into the culture chamber of our bioreactor for six weeks in vitro culture.

The cartilage yield in our recirculating bioreactor was 75-85%. The outcome was better than the 65-75% in the spinner flask bioreactor (shear stress only) and 17% in static culture. In addition, there was a significant difference in the cell morphology and zonal organisation among the three methods of culture; the engineered cartilage in the recirculating bioreactor presented many more characteristics of native articular cartilage.

If the environment of culture provides the shear stress and hydrodynamic pressure simultaneously, the composition of the engineered cartilage resembles native articular cartilage, including their ECM composition, cell distribution, zonal organisation and mechanical properties.