Department of Biomedical Engineering, Eindhoven University of Technology; Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology.
Department of Biomedical Engineering, Eindhoven University of Technology.
J Vis Exp. 2020 Dec 10(166). doi: 10.3791/61824.
The use of resorbable biomaterials to induce regeneration directly in the body is an attractive strategy from a translational perspective. Such materials induce an inflammatory response upon implantation, which is the driver of subsequent resorption of the material and the regeneration of new tissue. This strategy, also known as in situ tissue engineering, is pursued to obtain cardiovascular replacements such as tissue-engineered vascular grafts. Both the inflammatory and the regenerative processes are determined by the local biomechanical cues on the scaffold (i.e., stretch and shear stress). Here, we describe in detail the use of a custom-developed bioreactor that uniquely enables the decoupling of stretch and shear stress on a tubular scaffold. This allows for the systematic and standardized evaluation of the inflammatory and regenerative capacity of tubular scaffolds under the influence of well-controlled mechanical loads, which we demonstrate on the basis of a dynamic co-culture experiment using human macrophages and myofibroblasts. The key practical steps in this approach-the construction and setting up of the bioreactor, preparation of the scaffolds and cell seeding, application and maintenance of stretch and shear flow, and sample harvesting for analysis-are discussed in detail.
从转化医学的角度来看，使用可吸收生物材料直接在体内诱导再生是一种很有吸引力的策略。这种材料在植入后会引发炎症反应，这是随后材料吸收和新组织再生的驱动力。这种策略也被称为原位组织工程，旨在获得心血管替代品，如组织工程血管移植物。炎症和再生过程都取决于支架上的局部生物力学线索（即拉伸和切变应力）。在这里，我们详细描述了使用定制开发的生物反应器的方法，该生物反应器能够独特地分离管状支架上的拉伸和切变应力。这使得在受控机械负荷的影响下，系统地和标准化地评估管状支架的炎症和再生能力成为可能，我们基于使用人巨噬细胞和肌成纤维细胞的动态共培养实验来证明这一点。该方法的关键实际步骤——生物反应器的构建和设置、支架和细胞接种的准备、拉伸和切变流的应用和维持以及用于分析的样本采集——都进行了详细讨论。
