Three-dimensional printing of stem cell-laden hydrogels submerged in a hydrophobic high-density fluid
Three-dimensional printing of stem cell-laden hydrogels submerged in a hydrophobic high-density fluid
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DOI：
10.1088/1758-5082/5/1/015003
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发表时间：
2013-03-01
期刊：
影响因子：
9
通讯作者：
Fischer, Horst
中科院分区：
文献类型：
--
作者：
Campos, Daniela F. Duarte;Blaeser, Andreas;Fischer, Horst
在过去的十年里，生物打印技术已经开始为再生医学和器官移植提供重要的组织工程策略。过去方法的主要缺点是材料印刷设备和基材组合不佳或不充分，以及印刷构件的相对较小尺寸。在这里，我们假设，当浸泡在全氟三丁胺(C12F27N)(一种疏水的高密度流体)中时，可以打印出携带细胞的水凝胶，并且这些放置在三维结构中的细胞仍然可以存活，从而允许细胞增殖和产生细胞外基质。人骨髓间充质干细胞和MG-63细胞被包裹在琼脂糖水凝胶中，然后在高密度氟碳支撑的三维结构中以高纵横比打印。制作了各种形状和大小的三维结构，并在六个多月的时间里保持稳定。活/死染色和DAPI染色显示，打印过程后24小时以及培养21天后细胞仍有活力。第14天和第21天的组织学和免疫组织化学分析显示，活细胞有明显的基质生成和增殖迹象。因此，在培养的两周内，打印的凝胶的压缩强度值增加，这表明未来在再生医学中的应用是令人鼓舞的结果。
Over the last decade, bioprinting technologies have begun providing important tissue engineering strategies for regenerative medicine and organ transplantation. The major drawback of past approaches has been poor or inadequate material-printing device and substrate combinations, as well as the relatively small size of the printed construct. Here, we hypothesise that cell-laden hydrogels can be printed when submerged in perfluorotributylamine (C12F27N), a hydrophobic high-density fluid, and that these cells placed within three-dimensional constructs remain viable allowing for cell proliferation and production of extracellular matrix. Human mesenchymal stem cells and MG-63 cells were encapsulated into agarose hydrogels, and subsequently printed in high aspect ratio in three dimensional structures that were supported in high density fluorocarbon. Three-dimensional structures with various shapes and sizes were manufactured and remained stable for more than six months. Live/dead and DAPI stainings showed viable cells 24 h after the printing process, as well as after 21 days in culture. Histological and immunohistochemical analyses after 14 and 21 days revealed viable cells with marked matrix production and signs of proliferation. The compressive strength values of the printed gels consequently increased during the two weeks in culture, revealing encouraging results for future applications in regenerative medicine.