Department of Biomedical Engineering, St. Louis University, St. Louis, MO 63103, USA.
J Tissue Eng Regen Med. 2010 Dec;4(8):619-27. doi: 10.1002/term.277.
Scientists are always looking for new tools to better mimic human anatomy and physiology, especially to study chronic respiratory disease. Airway remodelling is a predominant feature in asthma and occurs in conjunction with chronic airway inflammation. Both the inflammatory and repair processes alter the airway wall which is marked by anatomical, physiological and functional changes. A tissue-engineered model of bronchiole remodelling presents a novel approach to investigating the initiation and progression of airway remodelling. By developing a unique bioreactor system, cylindrical-shaped bronchioles constructed from well-characterized human lung primary cells have been engineered and examined with a much greater control over experimental variables. We have grown human bronchioles composed of fibroblasts, airway smooth muscle cells, small airway epithelial cells and extracellular matrices. The various cell types are in close proximity to one another for cell-cell signalling and matrix interactions. The cylindrical geometry of the tissue applies radial distension for mechanotransduction and the air interface provides a natural environment for the epithelial cells. Optimal cell density, extracellular matrix concentration and media composition were determined. Immunohistochemistry verified bronchiole phenotypic stability. Quiescence was gauged by protein expression which verified a change in phenotype after the initial fabrication stage and implementation of the air interface. A fabrication timeline was devised for repeatable bronchiole fabrication and to understand tissue contraction and cell-seeding duration. The stability of the bronchiole structures and their cellular composition lends these bronchioles to study cell-cell interactions and remodelling events while maintaining in vivo geometrical dimensions and relationships.
科学家一直在寻找新的工具来更好地模拟人体解剖结构和生理学,特别是研究慢性呼吸道疾病。气道重塑是哮喘的主要特征,与慢性气道炎症同时发生。炎症和修复过程都会改变气道壁,从而导致解剖、生理和功能的变化。支气管重塑的组织工程模型为研究气道重塑的发生和进展提供了一种新的方法。通过开发独特的生物反应器系统,使用经过充分表征的人肺原代细胞构建的圆柱形支气管已经被工程化,并可以更好地控制实验变量进行检查。我们已经培养了由成纤维细胞、气道平滑肌细胞、小气道上皮细胞和细胞外基质组成的人支气管。各种细胞类型彼此靠近,以便进行细胞间信号传递和基质相互作用。组织的圆柱形状适用于机械转导的径向扩张,气界面为上皮细胞提供了自然环境。确定了最佳细胞密度、细胞外基质浓度和培养基组成。免疫组织化学验证了支气管的表型稳定性。通过蛋白质表达来衡量静止状态,验证了初始制造阶段和空气界面实施后表型的变化。制定了制造时间线,以实现可重复的支气管制造,并了解组织收缩和细胞接种持续时间。支气管结构及其细胞组成的稳定性使这些支气管能够研究细胞-细胞相互作用和重塑事件,同时保持体内几何尺寸和关系。