Three-Dimensional Bioprinting of Decellularized Extracellular Matrix-Based Bioinks for Tissue Engineering

Abstract

Three-dimensional (3D) bioprinting is one of the most promising additive manufacturing technologies for fabricating various biomimetic architectures of tissues and organs. In this context, the bioink, a critical element for biofabrication, is a mixture of biomaterials and living cells used in 3D printing to create cell-laden structures. Recently, decellularized extracellular matrix (dECM)-based bioinks derived from natural tissues have garnered enormous attention from researchers due to their unique and complex biochemical properties. This review initially presents the details of the natural ECM and its role in cell growth and metabolism. Further, we briefly emphasize the commonly used decellularization treatment procedures and subsequent evaluations for the quality control of the dECM. In addition, we summarize some of the common bioink preparation strategies, the 3D bioprinting approaches, and the applicability of 3D-printed dECM bioinks to tissue engineering. Finally, we present some of the challenges in this field and the prospects for future development.

Keywords: 3D bioprinting; bioink; decellularized extracellular matrix; tissue engineering.

Figure 1

Schematic illustration highlighting the various decellularized extracellular matrix (dECM) preparation methods, and the bioink preparation strategies and applications of 3D bioprinting dECM-based bioinks in tissue engineering.

Figure 2

Representative illustration of extracellular matrix (ECM) compositional layout indicating cellular engagement with ECM biomolecules and primary components of general ECM space [28].

Figure 3

Strategies for preparing decellularized bioinks. (A) Illustration of cdECMMA bioink formulation containing cells and three-dimensional (3D) bioprinting process using cell-laden cdECMMA bioink: (i) Preparation of cdECMMA; and (ii) schematic diagram of preparation mechanism of cdECMMA [23]; (B) Schematic representation of the preparation of dECM powder–based bioink (i) and its application to bioprinting (ii) [25].

Figure 4

Schematic diagram of three representative 3D bioprinting technology devices: (A) extrusion bioprinting; (B) inject bioprinting; (C) DLP bioprinting.

Figure 5

3D bioprinting of biocompatible and functional meniscus constructs using meniscus-derived bioink: (i) decellularization process of meniscus; (ii) rheological properties of me-dECM bioink and COL bioink; (iii) CAD-based 3D bioprinting of diverse meniscus constructs of rabbit, beagle, and human models [138].

Figure 6

A typical example of liver-derived dECM bioink for 3D printing application: (i) digital image of fresh porcine liver/liver dECM/lyophilized liver dECM; (ii) schematic of DLP-based 3D printer; (iii) designed liver microtissue model and DLP printing results [68].

Figure 7

Two examples of skin-derived dECM bioinks for 3D printing applications. (A) Skin-derived bioink formulation and its properties analysis: (i) S-dECM bioink preparation process; (ii) quantitative analyses of dECM bioink, including collagen, GAGs, elastin, hyaluronic acid, and DNA; (iii) sol-gel transition of dECM bioink; and (iv) printability test of dECM bioink [142]; (B) Structure of the 3D-printed construct using skin bioink and gene expression: (i) cell-laden 3D scaffold; and (ii) changes in gene expression in the 3D-printed cell-laden construct [80].

Figure 8

Two examples of cardiac-derived dECM bioinks for 3D printing applications. (A) Schematic illustration of a new cardiac-derived dECM bioink for 3D printing: (i) schematic illustration of a two-step cross-linking mechanism that applies concurrent cross-linking of vitamin B2-induced covalent cross-linking and thermal cross-linking; (ii) 3D printing and cross-linking; and (iii) digital image of the scaffold [144]; (B) Schematic depicting the stages starting with the preparation of the hdECM bioink to fabrication of the cell-laden EHT: (i) development of the hdECM bioink; and (ii) fabrication of the cardiomyocyte-laden EHT using a 3D bioprinter [145].

Figure 9

Schematic of research strategy: (i) schematic diagram of manufacturing coaxial vascular device and materials; (ii) schematic diagram of the coaxial blood vessel [146].

Figure 10

Preparation of KdECM and KdECMMA-based bioink formulations: changes in gene expression in the 3D-printed cell-laden construct: (i) gross images of the decellularization process: (a) normal kidney, (b) SDS treatment for 36 h, (c) Triton X-100 treatment for 24 h, and (d) washing in saline for 72 h; (ii) schematic illustration of a photo-cross-linkable kidney-specific ECM hydrogel; (iii) photography of KdECMMA before and after UV cross-linking; (iv) printing code and gross images of the printed KdECMMA-based constructs [147].