人体软骨组织制作采用三维喷墨打印技术
Summary
本文介绍的方法显示了如何将一个商用喷墨打印机与紫外线同时聚合生物印刷。该打印机能够构建三维组织结构，细胞和生物材料。这里展示的研究构建了一个3D neocartilage。
Abstract
生物印刷，这是基于热喷墨印刷，是最有吸引力的应用技术在组织工程和再生医学的领域之一。与数字控制单元，支架和生长因子可以精确地沉积到所需的二维（2D）和三维（3D）位置迅速。因此，这种技术是制造组织模仿他们的母语解剖结构的理想方法。为了与原生地带性的组织，细胞外基质成分（ECM）和机械性能工程师软骨，我们开发了使用商用喷墨打印机能够进行3D软骨组织工程同时进行光聚合生物印刷一个平台。人软骨细胞悬浮于聚（乙二醇）二丙烯酸酯（PEGDA），被印刷于通过层 – 层组装三维neocartilage施工。印刷细胞固定在原来的位置存放，由surro支持unding脚手架在同步光聚合。印刷组织的机械性能类似于天然软骨。相对于传统的组织的制造，这需要较长的紫外线曝光，在印刷单元和同步光聚合的存活率显著更高。印刷neocartilage展现了出色的糖胺聚糖（GAG）和Ⅱ型胶原的生产，这与基因的表达相一致。因此，这个平台是理想的精确细胞分布和安排解剖组织工程。
Introduction
生物印刷基于热喷墨印刷是最有前途的应用技术在组织工程和再生医学的领域之一。数字控制和高吞吐量的打印头单元，支架和生长因子可以精确地沉积到所需的二维（2D）和三维（3D）位置的迅速发展。采用该技术在组织工程和再生医学1-9许多成功的应用已经实现。在本文中，生物印刷一个平台，建立与修改后的惠普（HP）DESKJET 500热喷墨打印机和同步光聚合体系。由聚（乙二醇）（PEG）配制合成的水凝胶已经表明维持软骨细胞的生存力和促进软骨ECM生成10,11的容量。此外，光交联聚乙二醇在水中具有低粘度，这使得它非常适合同时聚合物：水溶性高在生物印刷3D化。在本文中，人类软骨细胞悬浮在聚（乙烯）乙二醇二丙烯酸酯（PEGDA; 3,400兆瓦）的精确打印，构建neocartilage层 – 层与1,400 dpi的三维解析。观察到沉积的细胞在三维支架材料中均匀分布，即生成软骨组织具有优异的机械性能和增强的ECM生成。与此相反，在手动制造的凝胶，而不是其最初沉积仓的底部由于较慢的聚合骨架，而导致不均匀的软骨形成培养后2,3蓄积的细胞。
Protocol
Representative Results
Discussion
这种3D生物印刷系统和同步光聚合产能提供比使用注射器挤出蜂窝海藻酸钠水凝胶16骨软骨缺损的原位印刷的最佳先前报道的方法一个显著更高的打印分辨率。较高的打印分辨率是特别关键的软骨组织工程恢复解剖软骨带状组织。层 – 层组装过程中同时进行光聚合反应是至关重要的，以维持细胞和生物材料支架的三维结构的精确沉积。微细加工与各印刷层也导致带状层之间的?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
作者要感谢来自纽约首都地区研究联盟资助的支持。
Materials
| HP Deskjet 500 thermal inkjet printer | Hewlett-Packard | C2106a | Discontinued. Purchased refurbished from internet vendor. |
| HP black ink cartridge | Hewlett-Packard | 51626a | |
| Ultraviolet lamp | UVP | B-100AP | |
| UV light meter | General Tools | UV513AB | |
| Zeiss LSM 510 laser scanning microscope | Carl Zeiss | LSM 510 | |
| Dulbeccos Modified Eagles Medium (DMEM) | Mediatech | 10-013 | |
| Penicillin-streptomycin-glutamine (PSG) | Invitrogen | 10378-016 | |
| Accutase cell dissociation reagent | Invitrogen | A11105-01 | |
| Phosphate buffered saline (PBS) | Invitrogen | 10010-023 | |
| Live/Dead viability/cytotoxicity Kit | Invitrogen | L-3224 | |
| Poly(ethylene glycol) diacrylate (PEGDA) | Glycosan Biosystems | GS700 | |
| Irgacure 2959 | Ciba Specialty Chemicals | I-2959 | |
| Human articular chondrocytes | Lonza | CC-2550 |
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