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

Journal article
Authors Peter Apelgren
E. Karabulut
Matteo Amoroso
A. Mantas
H. M. Avila
Lars Kölby
T. Kondo
G. Toriz
P. Gatenholm
Published in Acs Biomaterials Science & Engineering
Volume 5
Issue 5
Pages 2482-2490
ISSN 2373-9878
Publication year 2019
Published at Institute of Clinical Sciences, Department of Plastic Surgery
Pages 2482-2490
Language en
Keywords 3D-bioprinting, bacterial nanocellulose, aqueous counter collision, bioinks, neocartilage formation, mesenchymal stem-cells, cellulose nanofibers, acetobacter-xylinum, microbial cellulose, potential scaffold, implant material, blood-vessels, biocompatibility, chondrocytes, vascularizatio
Subject categories Biomaterials Science


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 X 5 X 1 mm(3)) 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 mm(2) observed after 30 days and 85.6 +/- 30.0 cells per mm(2) 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.

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