Soyoung Hong (a), Ji Seon Kim (a), Boyoung Jung (a), Chonghyun Won *(b), Changmo Hwang *(a, c)
Herein, three-dimensional (3D) bioprinting of engineered constructs with cell-laden biomaterials was investigated for the development of 3D tissue constructs in vitro. The present article proposes a simple coaxial-nozzle-based printing method using a one-step gelling gelatin bioink containing different cell types for vascular structure generation. First, a gelatin bioink prepolymer with a tyramine functional group was synthesized. To facilitate rapid gelation, polyethylene glycol (PEG) was introduced as a spacer between gelatin and tyramine. The gelatin–PEG–tyramine (GPT) prepolymer underwent enzymatic crosslinking, which yielded a higher gelation rate of up to 4.24 ± 0.08 s. Second, one-step bioprinting of a cell-laden tubular structure was demonstrated using a coaxial type extruder and the GPT bioink with human umbilical vein endothelial cells (HUVECs) with or without human dermal fibroblasts (HDFs). The printed no-cell GPT tube was demonstrated to possess a perfusable vascular structure. The extruded tube with HUVECs-in-GPT sheath configuration resulted in an endothelial cell-lined hollow structure and was maintained for up to 8 days in vitro. Additionally, the coaxially extruded tube with HUVECs-in-core (gelatin) and HDFs-in-GPT sheath (GPT) configuration exhibited a distribution of these two cell types along the tube axis. In the current study, it was demonstrated that a radial distribution of multiple vascular cells can be simply achieved using a synthetic GPT bioink combined with a coaxial nozzle printing system, serving as a proof-of-concept for one-step generation of vascular constructs. The rapid gelling bioink prepolymer, in combination with a coaxial bioprinter nozzle mechanism, has great potential for the development of designed, printed, and organized 3D tissue architecture vascularization.