In Vivo Human Cartilage Formation in Three-Dimensional Bioprinted Constructs with a Novel Bacterial Nanocellulose Bioink

Abstract

Bacterial nanocellulose (BNC) is a 3D network of nanofibrils exhibiting excellent biocompatibility. Here, we present the aqueous counter collision (ACC) method of BNC disassembly to create bioink with suitable properties for cartilage-specific 3D-bioprinting. BNC was disentangled by ACC, and fibril characteristics were analyzed. Bioink printing fidelity and shear-thinning properties were evaluated. Cell-laden bioprinted grid constructs (5 × 5 × 1 mm3) containing human nasal chondrocytes (10 M mL–1) were implanted in nude mice and explanted after 30 and 60 days. Both ACC and hydrolysis resulted in significantly reduced fiber lengths, with ACC resulting in longer fibrils and fewer negative charges relative to hydrolysis. Moreover, ACC-BNC bioink showed outstanding printability, postprinting mechanical stability, and structural integrity. In vivo, cell-laden structures were rapidly integrated, maintained structural integrity, and showed chondrocyte proliferation, with 32.8 ± 13.8 cells per mm2 observed after 30 days and 85.6 ± 30.0 cells per mm2 at day 60 (p = 0.002). Furthermore, a full-thickness skin graft was attached and integrated completely on top of the 3D-bioprinted construct. The novel ACC disentanglement technique makes BNC biomaterial highly suitable for 3D-bioprinting and clinical translation, suggesting cell-laden 3D-bioprinted ACC-BNC as a promising solution for cartilage repair.

3D Bioprinting of Lignocellulosic Biomaterials

“…[98] Strong nanofibrils and high-water retention capacity of MBC make it attractive for 3D printing; however, the disentanglement of intricated MBC fibrillar networks is one major challenge toward 3D printing of MBC for biomaterial applications which if not correctly disentangled can block the printer nozzles. Using the aqueous counter collision technique which is based on collision energy of dual water jets, Apelgren et al [99] disintegrated bacterial cellulose nanocrystals into fibrils. The biomaterial ink developed based on the aqueous counter collision (ACC) treated MBC showed excellent printability with structural integrity and postprint stability ( Figure 10II).…”

“…Structures bioprinted with high resolution and stability using bacterial cellulose after disassembly. [99] Reproduced with permission. [99] Copyright 2019, American Chemical Society.…”

Nanocellulose-Based Inks for 3D Bioprinting: Key Aspects in Research Development and Challenging Perspectives in Applications—A Mini Review

“…The surface chemistry of the BNC scaffold was also tailored by a coating of collagen to improve cell adhesion, growth, and differentiation [ 67 ]. The cell-laden 3D-bioprinting of BNC-based scaffolds has brought up promising solutions for tissue repair [ 71 , 72 ]. A recent review by A. Sionkowska et al presented recent advances of the most studied medical applications of BNC [ 73 ].…”

Study on 3D printing technology and mechanical properties of a nano‐enhanced composite hydrogel bio‐ink

“…A 3D scaffold must have controllable pore size, high porosity, and good pore connectivity [3–5] because it needs to simulate the tissue extracellular matrix [6, 7]; thus proper choice of bio‐inks [8] and parameter optimisation of printing [2, 9] are two focuses of 3D bioprinting. In previous studies, natural polymers (collagen [10], silk fibroin [11], and nanocellulose [12, 13]) and synthetic polymers (polyvinyl alcohol [14] and polyethylene glycol [15]) hydrogel bio‐inks were successfully applied to 3D bioprinting and made some progress.…”