3D Bioprinting Engineering Artificial Respiratory Tract Tissue

拟议的项目适合优先领域：前沿卫生背景：需要开发和改进与人类相关的工具，使之能够研究人类生物学中尚未完全理解的机制。目前的活体模型几乎完全使用非灵长类动物，这受到人类和动物系统之间频繁的相关性差的限制。理想的模型将允许在不同的实验条件下直接询问和比较基因等同的人体组织。在体外培养人体组织方面的进展，导致了体外人体器官培养的发展。到目前为止，这项研究的重点是生成用于移植的人体器官。然而，开发的技术也为人类生物学的体外研究提供了革命性的潜力。虽然脱细胞支架有望用于移植，但人类或动物组织生成支架的要求限制了该技术在其他领域的广泛应用，如科学发现。3D生物打印是创建空间控制的细胞模式的过程，在该过程中可以复制生物组织的行为。这一技术理想地扩展到打印完整的、可存活的器官，用于体外模型、组织修复和器官移植。打印可存活的器官目前还不可能，因为器官非常复杂。大多数打印的组织构建物不能存活很长时间，可打印的细胞负载生物墨水和打印后构建体的灌流仍然是悬而未决的问题。尽管面临挑战，但仍通过3D生物打印生产出了具有功能的微型器官，有望进一步发展。研究计划：该项目的目标是开发和优化可用于体外人类呼吸道器官培养的工程化人工支架。我们建议利用3D生物打印技术的进步，研究能够在试管中生长器官的方法，最终目标是显著减少/完全取代在研究中使用动物器官。具体而言，研究方案将包括：(1)印刷人造人体上皮组织1.2影响概要(最多1500个字符)1.3数据共享(最多2000个字符)2第一步是开发可打印的水凝胶和用于制造呼吸道组织的细胞生物墨水。由成纤维细胞基质和上皮细胞组成的多细胞结构的人工气道将被打印和培养，以在气液界面模型中生长完全分化的上皮细胞。通过组织学分析和生物标志物分析，将研究打印支架内的细胞自组装，并将其与来自捐赠者组织的人类肺支架进行比较。(Ii)3D打印微型整体器官支架项目第二阶段将打印整合了呼吸道的人造肺组织，与人类细胞在人体整个器官支架上的自组装进行比较。基于组织学、转录学和蛋白质生物标记物的分析将被用来比较使用微型支架产生的人造组织和整个捐赠者组织。(Iii)微型人造组织的功能研究这项研究中产生的组织将使用先前描述的组织功能介质进行测试(例如，在肺组织模型中辛伐他汀抑制上皮细胞的呼吸道粘液产生，在肠道组织模型中通过白细胞介素1β抑制乙酰胆碱诱导的肠道运动)。将根据第(1)和(2)项的结果界定所调查的具体功能调解人。

The proposed project fits in Priority Area: Leading Edge HealthcareBackground:There is a need to develop and improve human relevant tools enabling the study of mechanisms not yet fully understood in human biology. Current in vivo models almost exclusively utilise non primate animals which are limited by frequent poor correlation between human and animal systems. The ideal model would allow the direct interrogation and comparison of genetically equivalent human tissues under different experimental conditions. Advances in growing human tissues ex vivo, have led to the development of ex vivo human organ culture. To date this has focused on generating human organs for transplantation. However, the techniques developed also offer the potential to revolutionise in vitro studies of human biology. Whilst decellularised scaffolds hold promise for transplantation, the requirement for human or animal tissue to generate scaffolds limits the broad scale application of the technology to other areas such as science discovery.3D bio-printing is the process of creating spatially-controlled cell patterns, in which the behaviour of biological tissues can be reproduced. This ideally extends to printing complete, viable organs for in vitro model, tissue repair and organ transplant. Printing viable organs is not possible as of yet, as organs are very complex. Most printed tissue constructs are not viable for very long, printable cell-laden bioink and perfusion of the construct after printing are still open issues. Despite challenges, the functioning micro- organs have been produced by 3D bioprinting, promising for further development.Research Plan:The aim of this project is to develop and optimise engineered artificial scaffolds amenable to ex vivo human respiratory tract organ culture. We propose to investigate approaches to enable the growth of organs in test tubes, by making use of advances in 3D bioprinting technologies with an ultimate goal to significantly reduce/entirely replace the use of animal organs in research. Specifically the programme of research would consist of:(i) Printing artificial human epithelial tissue1.2 IMPACT SUMMARY (Up to 1500 characters)1.3 DATA SHARING (Up to 2000 characters)2The first step is to develop printable hydrogels and cell loaded bio-ink for fabricating airway tissue. Artificial airway with multicellular structure consisting of fibroblast stroma with epithelium will be printed and cultured for growing a fully differentiated epithelium in air-liquid interface models. Cellular self-assembly within printed scaffold will be studied and compared with human lung scaffolds from donor tissue through histological analysis and biomarker profiling.(ii) 3D printing miniaturised whole organ scaffoldsThe 2nd stage of the project will print artificial lung tissue with integration of airway, compared with human cell self-assembly on human whole organ scaffolds. Histological, transcriptomic and protein biomarker based profiling would be used to compare artificial tissue generated using miniaturised scaffolds to whole donor tissue.(iii) Functional studies of miniaturised artificial tissueThe tissue generated within this study would be tested using previously described mediators of tissue function (e.g. inhibition of airway mucus production of epithelial cells by simvastatin in a lung tissue model, or inhibition of acetylcholine induced intestinal motility by interleukin 1 beta in in a gut model). The specific functional mediators investigated would be defined based on the outcome of (i) and (ii).