Dual crosslinking silk fibroin/pectin-based bioink development and the application on neural stem/progenitor cells spheroid laden 3D bioprinting

Hao-Wei Lee; Ko-Ting Chen; Yi-Chen Ethan Li; Yu-Chun Yeh; Chao-Ying Chiang; I-Chi Lee

doi:10.1016/j.ijbiomac.2024.131720

Dual crosslinking silk fibroin/pectin-based bioink development and the application on neural stem/progenitor cells spheroid laden 3D bioprinting

Int J Biol Macromol. 2024 Apr 25;269(Pt 2):131720. doi: 10.1016/j.ijbiomac.2024.131720. Online ahead of print.

Authors

Hao-Wei Lee¹, Ko-Ting Chen², Yi-Chen Ethan Li³, Yu-Chun Yeh¹, Chao-Ying Chiang³, I-Chi Lee⁴

Affiliations

¹ Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan.
² Department of Neurosurgery, Chang Gung Memorial Hospital at Linkou, Taoyuan, Taiwan.
³ Department of Chemical Engineering, Feng Chia University, Taichung 40724, Taiwan.
⁴ Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan. Electronic address: iclee@mx.nthu.edu.tw.

PMID: 38677692
DOI: 10.1016/j.ijbiomac.2024.131720

Abstract

The human nervous system is an incredibly intricate physiological network, and neural cells lack the ability to repair and regenerate after a brain injury. 3-dimensional (3D) bioprinting technology offers a promising strategy for constructing biomimetic organ constructs and in vitro brain/disease models. The bioink serves as a pivotal component that emulates the microenvironment of biomimetic construct and exerts a profound influence on cellular behaviors. In this study, a series of mechanically adjustable and dual crosslinking bioinks were developed using photocrosslinkable methacrylated silk fibroin (SilMA) in combination with the ionic crosslinking material, pectin, or pectin methacryloyl (PecMA) with silk fibroin (SF) supplementation. SilMA/pectin exhibited superior properties, with SilMA providing biocompatibility and adjustable mechanical properties, while the addition of pectin enhanced printability. The porous structure supported neural cell growth, and 15 % SilMA/0.5 % pectin bioinks displayed excellent printability and shape fidelity. Neural stem/progenitor cells (NSPCs)-loaded bioinks were used to construct a 3D brain model, demonstrating sustained vitality and high neuronal differentiation without the need for growth factors. The SilMA/pectin bioinks demonstrated adjustable mechanical properties, favorable biocompatibility, and an environment highly conducive to neural induction, offering an alternative approach for neural tissue engineering applications or in vitro brain models.

Keywords: 3D bioprinting; Bioink; In vitro brain model; Neural stem/progenitor cells (NSPCs); Pectin; Silk fibroin/methacrylated silk fibroin (SilMA).