用于模拟羊水运动影响的动态流动胎儿膜器官芯片系统。
A dynamic flow fetal membrane organ-on-a-chip system for modeling the effects of amniotic fluid motion.
机构信息
Department of Electrical and Computer Engineering, Texas A&M University, College Station, TX, USA.
Division of Basic Science and Translational Research, Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, USA.
出版信息
Biomed Microdevices. 2024 Jul 4;26(3):32. doi: 10.1007/s10544-024-00714-1.
Fetal membrane (amniochorion), the innermost lining of the intrauterine cavity, surround the fetus and enclose amniotic fluid. Unlike unidirectional blood flow, amniotic fluid subtly rocks back and forth, and thus, the innermost amnion epithelial cells are continuously exposed to low levels of shear stress from fluid undulation. Here, we tested the impact of fluid motion on amnion epithelial cells (AECs) as a bearer of force impact and their potential vulnerability to cytopathologic changes that can destabilize fetal membrane functions. A previously developed amnion membrane (AM) organ-on-chip (OOC) was utilized but with dynamic flow to culture human fetal amnion membrane cells. The applied flow was modulated to perfuse culture media back and forth for 48 h to mimic fluid motion. A static culture condition was used as a negative control, and oxidative stress (OS) condition was used as a positive control representing pathophysiological changes. The impacts of fluidic motion were evaluated by measuring cell viability, cellular transition, and inflammation. Additionally, scanning electron microscopy (SEM) imaging was performed to observe microvilli formation. The results show that regardless of the applied flow rate, AECs and AMCs maintained their viability, morphology, innate meta-state, and low production of pro-inflammatory cytokines. E-cadherin expression and microvilli formation in the AECs were upregulated in a flow rate-dependent fashion; however, this did not impact cellular morphology or cellular transition or inflammation. OS treatment induced a mesenchymal morphology, significantly higher vimentin to cytokeratin 18 (CK-18) ratio, and pro-inflammatory cytokine production in AECs, whereas AMCs did not respond in any significant manner. Fluid motion and shear stress, if any, did not impact AEC cell function and did not cause inflammation. Thus, when using an amnion membrane OOC model, the inclusion of a dynamic flow environment is not necessary to mimic in utero physiologic cellular conditions of an amnion membrane.
胎膜(羊膜绒毛膜)是子宫腔内最内层的衬里,围绕着胎儿并包裹着羊水。与单向血流不同,羊水会微妙地前后晃动,因此,最内层的羊膜上皮细胞会不断受到来自流体波动的低水平剪切力的影响。在这里,我们测试了流体运动对羊膜上皮细胞(AEC)的影响,因为它们是力冲击的载体,以及它们对可能使胎膜功能不稳定的细胞病理变化的潜在脆弱性。我们使用了之前开发的羊膜膜(AM)器官芯片(OOC),但采用动态流动来培养人胎儿羊膜膜细胞。施加的流动被调节为在 48 小时内来回灌注培养基,以模拟流体运动。静态培养条件用作阴性对照,氧化应激(OS)条件用作代表病理生理变化的阳性对照。通过测量细胞活力、细胞转化和炎症来评估流体运动的影响。此外,还进行了扫描电子显微镜(SEM)成像以观察微绒毛形成。结果表明,无论施加的流速如何,AEC 和 AMCs 都保持其活力、形态、固有状态和低水平促炎细胞因子的产生。E-钙粘蛋白在 AEC 中的表达和微绒毛形成呈流速依赖性上调;然而,这并没有影响细胞形态、细胞转化或炎症。OS 处理诱导 AEC 出现间充质形态,显著增加波形蛋白与细胞角蛋白 18(CK-18)的比值,并增加促炎细胞因子的产生,而 AMCs 没有任何显著反应。如果存在流体运动和剪切力,则不会影响 AEC 细胞功能,也不会引起炎症。因此,在使用羊膜膜 OOC 模型时,不需要包含动态流动环境来模拟羊膜膜在子宫内的生理细胞条件。