Coaxial Printing of Double-layered and Free-standing Blood Vessel Analogues Without Ultraviolet Illumination for High-volume Vascularised Tissue

Human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) were coaxially and continuously extruded without ultraviolet illumination using a microfluidic-based nozzle. Type I collagen (3 mg ml) containing HUVECs and a crosslinking reagent (100 mM CaCl) were supplied as the core material. A mixture of 3 mg ml of type I collagen (25%) and 1.8% weight volume of sodium alginate (75%) was provided as the shell layer material surrounding the core material. The HUVECs were well proliferated at the core and reshaped into a monolayer formation along the axial direction of the scaffold. The HASMCs showed more than 90% cell viability in the shell layer. Fluorescent beads were passed through the inside channel of the scaffold with the HUVEC core and HASMC shell using an in-house connector. This double-layered scaffold showed higher angiogenesis in growth factor-free medium than the scaffold with only a HUVEC core. The HASMCs in the shell layer affected angiogenesis, extracellular matrix secretion, and outer diameter. The proposed technique could be applied to three-dimensional bioprinting for the production of high-volume vascularised tissue.