Kappa-Carrageenan-Based Dual Crosslinkable Bioink for Extrusion Type Bioprinting

Abstract

Bioink based 3D bioprinting is a promising new technology that enables fabrication of complex tissue structures with living cells. The printability of the bioink depends on the physical properties such as viscosity. However, the high viscosity bioink puts shear stress on the cells and low viscosity bioink cannot maintain complex tissue structure firmly after the printing. In this work, we applied dual crosslinkable bioink using Kappa-carrageenan (κ-CA) to overcome existing shortcomings. κ-CA has properties such as biocompatibility, biodegradability, shear-thinning and ionic gelation but the difficulty of controlling gelation properties makes it unsuitable for application in 3D bioprinting. This problem was solved by synthesizing methacrylated Kappa-carrageenan (MA-κ-CA), which can be dual crosslinked through ionic and UV (Ultraviolet) crosslinking to form hydrogel using NIH-3T3 cells. Through MA substitutions, the rheological properties of the gel could be controlled to reduce the shear stress. Moreover, bioprinting using the cell-laden MA-κ-CA showed cell compatibility with enhanced shape retention capability. The potential to control the physical properties through dual crosslinking of MA-κ-CA hydrogel is expected to be widely applied in 3D bioprinting applications.

Recent Advances in Chemically-Modified and Hybrid Carrageenan-Based Platforms for Drug Delivery, Wound Healing, and Tissue Engineering

“…Table 1 summarizes the various functionalization of the carrageenan backbone using various types of monomers for different biomedical applications. Between them, the methacrylation of carrageenan, and the synthesis of methacrylate κ-carrageenan (KaMA), has attracted wide attention in the recent years [ 10 , 11 , 13 , 14 , 49 , 50 ]. According to Figure 1 A, by substituting the hydroxyl groups on κ-carrageenan with methacrylate groups, the methacrylation process occurred.…”

“…This hydrogel might be employed to reconstruct the bone–cartilage interface, where a natural gradient in the structural, mechanical, and cell interaction is present. Lim et al [ 10 ] applied a dual-crosslinkable bionic using methacrylated κ-carrageenan for soft TE. Bioprinting using the cell-laden (NIH-3T3 cells) hydrogel showed cell compatibility with an enhanced shape retention capability.…”

“…: ( A ) Schematic representation of the synthesis methacrylate κ-carrageenan. Reproduced with permission from [ 10 ]. ( B ) Assessment of dual-crosslinked hydrogel.…”

“…: ( A ) 3D incorporating NIH-3T3 cells in carrageenan hydrogel assay with DAPI/F-actin staining image at days seven and nine. Reproduced with permission from [ 10 ]. ( B ) Schematic outlining the design strategy to synthesize nanohybrid KaMA–GOPD hydrogels.…”

“…Additionally, carrageenans were employed to synthesize carrageenan-based hydrogels for various bio-applications including tissue engineering (TE) and drug delivery (DD) systems and regenerative medicine [ 9 ]. However, these applications have been limited due to the difficulty of controlling gelation properties, stability, and degradation rate in the physiological environment [ 10 ]. Various physical or chemical crosslinking methods allow the formation of networks with better properties to overcome these challenges.…”