Establishing a novel 3D printing bioinks system with recombinant human collagen

Abstract

Bioinks are one of the key elements in realizing three-dimensional (3D) bioprinting. However, bioinks prepared from conventional collagen are hindered to their further applications due to concerns of collagen purity, unstable mechanical properties, and low solubility under neutralized conditions. This study aimed to develop a reliable UV-curable bioink system from a novel water-soluble recombinant human collagen (RHC). RHC was modified by methacrylic anhydride (MAA) and later crosslinked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS) to obtain Pro-RHCMA. ¹H nuclear magnetic resonance (¹H NMR) confirmed the methacryloyl grafts, Fourier transform-infrared spectroscopy (FT-IR) illustrated the chemical crosslinking in producing the Pro-RHCMA. Internal morphology, mechanical properties and degradation of UV cured boinks were MAA and EDC/NHS modification-dependent. Photorheological properties and printability of the bioinks were determined. Cellular bioactivities were sustained within the printed bioinks, validating the bioinks biocompatibility in vitro. Finally, qRT-PCR revealed that the Pro-RHCMA bioinks provided a cell-friendly microenvironment for human umbilical vein endothelial cells (HUVECs) and human foreskin fibroblasts (HFFs), by supporting the expression of extracellular matrix (ECM) and angiogenesis-associated proteins, respectively. Taken together, this novel RHC-based bioink system shows great potential in tissue engineering and regenerative medicine.

Keywords: 3D bioprinting; Recombinant human collagen; UV-curable bioinks.