In Vivo Human Cartilage Formation in Three-Dimensional Bioprinted Constructs with a Novel Bacterial Nanocellulose Bioink
In Vivo Human Cartilage Formation in Three-Dimensional Bioprinted Constructs with a Novel Bacterial Nanocellulose Bioink
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DOI：
10.1021/acsbiomaterials.9b00157
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发表时间：
2019-05-01
影响因子：
5.8
通讯作者：
Gatenholm, Paul
中科院分区：
文献类型：
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作者：
Apelgren, Peter;Karabulut, Erdem;Gatenholm, Paul
细菌纳米纤维素（BNC）是一种具有良好生物相容性的纳米纤维三维网络。在这里，我们提出了BNC拆卸的水逆碰撞（ACC）方法，以创建具有适合软骨特异性3D生物打印特性的生物墨水。用ACC对BNC进行解缠结，并分析原纤维特征。评价了生物油墨打印保真度和剪切稀化特性。将含有人鼻软骨细胞（10 M mL（-1））的载有细胞的生物打印网格构建体（5 × 5 × 1 mm（3））植入裸鼠体内，并在30天和60天后接种。ACC和水解都导致纤维长度显著降低，ACC导致相对于水解的更长的原纤维和更少的负电荷。此外，ACC-BNC生物油墨显示出出色的可印刷性、印刷后机械稳定性和结构完整性。在体内，载有细胞的结构迅速整合，保持结构完整性，并显示软骨细胞增殖，30天后观察到32.8 +/- 13.8个细胞/mm 2，60天为85.6 +/- 30.0个细胞/mm 2（p = 0.002）。此外，将全层皮肤移植物连接并完全整合在3D生物打印结构的顶部。新的ACC解缠结技术使BNC生物材料非常适合3D生物打印和临床翻译，这表明载有细胞的3D生物打印ACC-BNC是软骨修复的一种有前途的解决方案。
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.