Department of Bioengineering, Clemson University, Clemson, SC, USA.
Gazes Cardiac Research Institute, Division of Cardiology, Department of Medicine, Medical University of South Carolina, USA.
J Biomech. 2023 Jan;147:111458. doi: 10.1016/j.jbiomech.2023.111458. Epub 2023 Jan 18.
Cardiac fibrosis is a key contributor to the onset and progression of heart failure and occurs from extracellular matrix accumulation via activated cardiac fibroblasts. Cardiac fibroblasts activate in response to mechanical stress and have been studied in the past by applying forces and deformations to three-dimensional, cell-seeded gels and tissue constructs in vitro. Unfortunately, previous stretching platforms have traditionally not enabled mechanical property assessment to be performed with an efficient throughput, thereby limiting the full potential of in vitro mechanobiology studies. We have developed a novel in vitro platform to study cell-populated tissue constructs under dynamic mechanical stimulation while also performing repeatable, non-destructive stress-strain tests in living constructs. Additionally, this platform can perform these tests across all constructs in a multi-well plate simultaneously, providing exciting potential for direct, functional readouts in future screening applications. In our pilot application, we showed that cyclically stretching cell-populated tissue constructs composed of murine cardiac fibroblasts within a 3D fibrin matrix leads to collagen accumulation and increased tissue stiffness over a three-day time course. Results of this study validate our platform's ability to apply mechanical loads to tissues while performing live mechanical analyses to observe cell-mediated tissue remodeling.
心肌纤维化是心力衰竭发生和进展的关键因素,它通过激活的心肌成纤维细胞导致细胞外基质的积累而发生。心肌成纤维细胞在受到机械应激时会被激活,过去曾通过在体外向三维细胞接种凝胶和组织构建体施加力和变形来研究这些细胞。不幸的是,传统的拉伸平台通常无法以高效的吞吐量进行机械性能评估,从而限制了体外力学生物学研究的全部潜力。我们开发了一种新的体外平台,可在动态机械刺激下研究细胞填充的组织构建体,同时还可以对活构建体进行可重复、非破坏性的应力-应变测试。此外,该平台可以同时在多孔板中的所有构建体上进行这些测试,为未来的筛选应用提供了直接、功能读数的令人兴奋的潜力。在我们的初步应用中,我们表明,在 3D 纤维蛋白基质中周期性地拉伸由小鼠心肌成纤维细胞组成的细胞填充组织构建体,会导致胶原蛋白的积累,并在三天的时间内增加组织的硬度。这项研究的结果验证了我们的平台在对组织施加机械负载的同时进行实时机械分析以观察细胞介导的组织重塑的能力。