Wan Weimin, Wang Xi, Zhang Rongtao, Li Yixuan, Wu Haonan, Liu Yiman, Zhang Fan, Liu Jia, Liu Guiquan, Zhou Lin, Wu Zhenhua, Mao Hongju, Yang Jian
State Key Laboratory of Component-Based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, China.
Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, China.
J Tissue Eng. 2025 Mar 31;16:20417314251328128. doi: 10.1177/20417314251328128. eCollection 2025 Jan-Dec.
By integrating 3D-inkjet bioprinting technology, differentiated human cells can be assembled into artificial lung tissue structure to achieve a rapid, efficient, and reproducible disease model construction process. Here, we developed a novel 3D-inkjet bioprinting-based method to construct artificial lung tissue structure (ALTs) for acute lung injury (ALI) disease modeling, research and application. It can also be used to study the role of relevant cells in the disease by adjusting the cell type and adapted to study the bio-functions of immune cells during the cell-cell interactions. Firstly, a series of process optimizations were done to mass-produce the alginate hydrogel microspheres (Alg) with a particle size of 262.63 ± 5 μm using a 3D bioprinter, then the type I collagen and polydopamine were deposited in turns to construct a cell adhesion layer on the surfaces of Alg (P-Alg) and the particle size was increased to 328.41 ± 3.81 μm. This platform exhibites good stability, timescale-dependent behavior, and long-term cell adhesion. Subsequently, several human cells including endothelial, epithelial, fibroblast, and even immune cells such as macrophages were adhered to P-Alg through rotational culture, leading to cell contractions and aggregation, subsequently formed ALTs or ALTs with macrophages (ALTs@M) with human alveolar-like structure. Finally, we successfully constructed an ALI model with lung barrier damage on ALTs using lipopolysaccharide stimulation in vitro, and comparison of secreted inflammatory factors between ALTs and ALTs@M. Results demonstrated that ALTs@M was more effective than ALTs in stimulating the inflammatory microenvironment of the lungs, providing a novel in vitro model for cellular interactions and human macrophage research. Altogether, this artificial lung tissue structure construction strategy using 3D-inkjet bioprinting technology allowed the flexible development of artificial lung tissue structures as potential disease models for preclinical studies.
通过整合3D喷墨生物打印技术，可将分化的人类细胞组装成人工肺组织结构，以实现快速、高效且可重复的疾病模型构建过程。在此，我们开发了一种基于3D喷墨生物打印的新方法来构建用于急性肺损伤（ALI）疾病建模、研究及应用的人工肺组织结构（ALTs）。它还可通过调整细胞类型用于研究相关细胞在疾病中的作用，并适用于研究细胞间相互作用过程中免疫细胞的生物功能。首先，进行了一系列工艺优化，使用3D生物打印机批量生产粒径为262.63±5μm的海藻酸盐水凝胶微球（Alg），然后依次沉积I型胶原蛋白和聚多巴胺，在Alg（P-Alg）表面构建细胞粘附层，粒径增大至328.41±3.81μm。该平台具有良好的稳定性、时间尺度依赖性行为和长期细胞粘附性。随后，包括内皮细胞、上皮细胞、成纤维细胞等几种人类细胞，甚至巨噬细胞等免疫细胞通过旋转培养粘附到P-Alg上，导致细胞收缩和聚集，随后形成具有人肺泡样结构的ALTs或带有巨噬细胞的ALTs（ALTs@M）。最后，我们在体外使用脂多糖刺激成功构建了具有肺屏障损伤的ALI模型，并比较了ALTs和ALTs@M之间分泌的炎症因子。结果表明，ALTs@M在刺激肺部炎症微环境方面比ALTs更有效，为细胞间相互作用和人类巨噬细胞研究提供了一种新的体外模型。总之，这种使用3D喷墨生物打印技术的人工肺组织结构构建策略允许灵活开发人工肺组织结构作为临床前研究的潜在疾病模型。
