Microfluidic-enhanced 3D bioprinting of aligned myoblast-laden hydrogels leads to functionally organized myofibers in vitro and in vivo
Microfluidic-enhanced 3D bioprinting of aligned myoblast-laden hydrogels leads to functionally organized myofibers in vitro and in vivo
复制标题
DOI：
10.1016/j.biomaterials.2017.03.026
复制
发表时间：
2017-07-01
期刊：
影响因子：
14
通讯作者：
Gargioli, Cesare
中科院分区：
文献类型：
--
作者：
Costantini, Marco;Testa, Stefano;Gargioli, Cesare
我们提出了一种基于创新的3D生物打印方法来制造具有功能形态的人工骨骼肌组织的新策略。该方法基于连接到同轴针挤出机的微流控打印头，用于高分辨率3D生物打印充满肌肉前体细胞(C2C12)的水凝胶纤维。为了促进成肌细胞分化，我们研制了一种定制的生物墨水，其中含有可光固化的半合成生物聚合物(聚乙二醇-纤维蛋白原)，将细胞封装成由排列的水凝胶纤维组成的3D结构。培养3-5天后，包裹的成肌细胞开始迁移和融合，在3D生物打印的纤维中形成多核肌管。所获得的肌管沿水凝胶纤维沉积方向高度排列，进一步揭示了成熟、肌瘤形成和功能。在免疫功能低下的小鼠背部皮下植入后，生物打印构建物在体内产生了有组织的人造肌肉组织。最后，我们展示了我们的微流控打印头允许设计三维多细胞组件，并对封装的细胞进行了精细的分隔。我们的结果表明，随着平行排列的长距离肌管的形成，肌源性分化增强。我们在此报道的方法代表了一种健壮和有效的方法来制造宏观人工肌肉，以扩大骨骼肌组织工程用于人类临床应用。(三)2017年提交人(S)。爱思唯尔有限公司出版。
We present a new strategy for the fabrication of artificial skeletal muscle tissue with functional morphologies based on an innovative 3D bioprinting approach. The methodology is based on a microfluidic printing head coupled to a co-axial needle extruder for high-resolution 3D bioprinting of hydrogel fibers laden with muscle precursor cells (C2C12). To promote myogenic differentiation, we formulated a tailored bioink with a photocurable semi-synthetic biopolymer (PEG-Fibrinogen) encapsulating cells into 3D constructs composed of aligned hydrogel fibers. After 3-5 days of culture, the encapsulated myoblasts started migrating and fusing, forming multinucleated myotubes within the 3D bioprinted fibers. The obtained myotubes showed high degree of alignment along the direction of hydrogel fiber deposition, further revealing maturation, sarcomerogenesis, and functionality. Following subcutaneous implantation in the back of immunocompromised mice, bioprinted constructs generated organized artificial muscle tissue in vivo. Finally, we demonstrate that our microfluidic printing head allows to design three dimensional multi-cellular assemblies with an exquisite compartmentalization of the encapsulated cells. Our results demonstrate an enhanced myogenic differentiation with the formation of parallel aligned long-range myotubes. The approach that we report here represents a robust and valid candidate for the fabrication of macroscopic artificial muscle to scale up skeletal muscle tissue engineering for human clinical application. (C) 2017 The Author(s). Published by Elsevier Ltd.