Inspired Materials & Stem-Cell Based Tissue Engineering Laboratory (IMSTEL), The University of Texas at El Paso, El Paso, Texas 79968, United States.
Department of Metallurgical, Materials, and Biomedical Engineering, M201 Engineering, The University of Texas at El Paso, 500 W. University Avenue, El Paso, Texas 79968, United States.
ACS Appl Mater Interfaces. 2022 May 18;14(19):21800-21813. doi: 10.1021/acsami.1c23883. Epub 2022 May 9.
In this study, we developed three-dimensional (3D) printed annular ring-like scaffolds of hydrogel (gelatin-alginate) constructs encapsulated with a mixture of human cardiac AC16 cardiomyocytes (CMs), fibroblasts (CFs), and microvascular endothelial cells (ECs) as cardiac organoid models in preparation for investigating the role of microgravity in cardiovascular disease initiation and development. We studied the mechanical properties of the acellular scaffolds and confirmed their cell compatibility as well as heterocellular coupling for cardiac tissue engineering. Rheological analysis performed on the acellular scaffolds showed the scaffolds to be elastogenic with elastic modulus within the range of a native heart tissue. The microstructural and physicochemical properties of the scaffolds analyzed through scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy-attenuated total reflectance (ATR-FTIR) confirmed the mechanical and functional stability of the scaffolds for long-term use in cell culture studies. HL-1 cardiomyocytes bioprinted in these hydrogel scaffolds exhibited contractile functions over a sustained period of culture. Cell mixtures containing CMs, CFs, and ECs encapsulated within the 3D printed hydrogel scaffolds exhibited a significant increase in viability and proliferation over 21 days, as shown by flow cytometry analysis. Moreover, via the expression of specific cardiac biomarkers, cardiac-specific cell functionality was confirmed. Our study depicted the heterocellular cardiac cell interactions, which is extremely important for the maintenance of normal physiology of the cardiac wall and significantly increased over a period of 21 days in . This 3D bioprinted "cardiac organoid" model can be adopted to simulate cardiac environments in which cellular crosstalk in diseased pathologies like cardiac atrophy can be studied and can further be used for drug cytotoxicity screening or underlying disease mechanisms.
在这项研究中，我们开发了一种三维（3D）打印的水凝胶（明胶-海藻酸盐）环形支架，其中封装了人心脏 AC16 心肌细胞（CMs）、成纤维细胞（CFs）和微血管内皮细胞（ECs）的混合物，作为心脏类器官模型，用于研究微重力在心血管疾病发生和发展中的作用。我们研究了无细胞支架的机械性能，并确认了它们对心脏组织工程的细胞相容性和异细胞偶联性。对无细胞支架进行的流变分析表明，支架具有弹性，弹性模量在天然心脏组织的范围内。通过扫描电子显微镜（SEM）和傅里叶变换衰减全反射红外光谱（ATR-FTIR）分析支架的微观结构和物理化学性质，证实了支架的机械和功能稳定性，可长期用于细胞培养研究。在这些水凝胶支架中生物打印的 HL-1 心肌细胞在培养过程中表现出持续的收缩功能。通过流式细胞术分析，包含 CMs、CFs 和 ECs 的细胞混合物在封装在 3D 打印水凝胶支架中 21 天内表现出显著的活力和增殖增加。此外，通过表达特定的心脏生物标志物，证实了心脏特异性细胞功能。我们的研究描述了异细胞心脏细胞相互作用，这对于维持心脏壁的正常生理功能非常重要，并且在 21 天的时间内显著增加。这种 3D 生物打印的“心脏类器官”模型可用于模拟心脏环境，研究心脏萎缩等疾病病理中的细胞串扰，并可进一步用于药物细胞毒性筛选或潜在疾病机制的研究。
