Anil Kumar Shweta, Alonzo Matthew, Allen Shane C, Abelseth Laila, Thakur Vikram, Akimoto Jun, Ito Yoshihiro, Willerth Stephanie M, Suggs Laura, Chattopadhyay Munmun, Joddar Binata
Inspired Materials & Stem-Cell Based Tissue Engineering Laboratory (IMSTEL), Department of Metallurgical, Materials and Biomedical Engineering, M201 Metallurgy Building, United States.
Department of Biomedical Engineering, The University of Texas at Austin, 110 Inner Campus Drive, Austin, Texas 78712, United States.
ACS Biomater Sci Eng. 2019 Sep 9;5(9):4551-4563. doi: 10.1021/acsbiomaterials.9b00505. Epub 2019 Aug 1.
In this study, fibrin was added to a photo-polymerizable gelatin-based bioink mixture to fabricate cardiac cell-laden constructs seeded with human induced pluripotent stem cell-derived cardiomyocytes (iPS-CM) or CM cell lines with cardiac fibroblasts (CF). The extensive use of platelet-rich fibrin, its capacity to offer patient specificity, and the similarity in composition to surgical glue prompted us to include fibrin in the existing bioink composition. The cell-laden bioprinted constructs were cross-linked to retain a herringbone pattern via a two-step procedure including the visible light cross-linking of furfuryl-gelatin followed by the chemical cross-linking of fibrinogen via thrombin and calcium chloride. The printed constructs revealed an extremely porous, networked structure that afforded long-term in vitro stability. Cardiomyocytes printed within the sheet structure showed excellent viability, proliferation, and expression of the troponin I cardiac marker. We extended the utility of this fibrin-gelatin bioink toward coculturing and coupling of CM and cardiac fibroblasts (CF), the interaction of which is extremely important for maintenance of normal physiology of the cardiac wall in vivo. This enhanced "cardiac construct" can be used for drug cytotoxicity screening or unraveling triggers for heart diseases in vitro.
在本研究中，将纤维蛋白添加到可光聚合的明胶基生物墨水混合物中，以制备接种有人诱导多能干细胞衍生的心肌细胞（iPS-CM）或心肌细胞系与心脏成纤维细胞（CF）的载有心脏细胞的构建体。富含血小板的纤维蛋白的广泛应用、其提供患者特异性的能力以及与手术胶水在成分上的相似性促使我们将纤维蛋白纳入现有的生物墨水成分中。载有细胞的生物打印构建体通过两步程序交联以保留人字形图案，该程序包括糠基明胶的可见光交联，随后通过凝血酶和氯化钙对纤维蛋白原进行化学交联。打印的构建体显示出极其多孔的网络结构，具有长期的体外稳定性。打印在片状结构内的心肌细胞表现出优异的活力、增殖能力以及心肌肌钙蛋白I标志物的表达。我们将这种纤维蛋白-明胶生物墨水的应用扩展到心肌细胞与心脏成纤维细胞的共培养和耦合，它们之间的相互作用对于维持体内心脏壁的正常生理功能极为重要。这种增强的“心脏构建体”可用于体外药物细胞毒性筛选或揭示心脏病的触发因素。
