Bacci Cydnee, Wong Vanessa, Barahona Victor, Merna Nick
Bioengineering Program, Fred DeMatteis School of Engineering and Applied Sciences, Hofstra University, Hempstead, NY, USA.
Microcirculation. 2021 Jan;28(1):e12659. doi: 10.1111/micc.12659. Epub 2020 Sep 29.
Preconditioning of endothelial cells from different vascular beds has potential value for re-endothelialization and implantation of engineered tissues. Understanding how substrate stiffness and composition affects tissue-specific cell response to shear stress will aid in successful endothelialization of engineered tissues. We developed a platform to test biomechanical and biochemical stimuli.
A novel polydimethylsiloxane-based parallel plate flow chamber enabled application of laminar fluid shear stress of 2 dynes/cm for 12 hours to microvascular cardiac and lung endothelial cells cultured on cardiac and lung-derived extracellular matrix. Optical imaging of cells was used to quantify cell changes in cell alignment. Analysis of integrin expression was performed using flow cytometry.
Application of fluid shear stress caused the greatest cell alignment in cardiac endothelial cells seeded on polystyrene and lung endothelial cells on polydimethylsiloxane. This resulted in elongation of the lung endothelial cells. α and β integrin expression decreased after application of shear stress in both cell types.
Substrate stiffness plays an important role in regulating tissue-specific endothelial response to shear stress, which may be due to differences in their native microenvironments. Furthermore, cardiac and lung endothelial cell response to shear stress was significantly regulated by the type of coating used.
对来自不同血管床的内皮细胞进行预处理对于工程组织的再内皮化和植入具有潜在价值。了解底物硬度和组成如何影响组织特异性细胞对剪切应力的反应将有助于工程组织成功实现内皮化。我们开发了一个平台来测试生物力学和生化刺激。
一种新型的基于聚二甲基硅氧烷的平行板流动腔，能够对培养在心脏和肺来源的细胞外基质上的微血管心脏和肺内皮细胞施加2达因/平方厘米的层流剪切应力，持续12小时。利用细胞的光学成像来量化细胞排列的变化。使用流式细胞术分析整合素表达。
施加流体剪切应力后，接种在聚苯乙烯上的心脏内皮细胞和接种在聚二甲基硅氧烷上的肺内皮细胞出现了最大程度的细胞排列。这导致肺内皮细胞伸长。在两种细胞类型中，施加剪切应力后α和β整合素表达均下降。
底物硬度在调节组织特异性内皮细胞对剪切应力的反应中起重要作用，这可能是由于它们天然微环境的差异所致。此外，心脏和肺内皮细胞对剪切应力的反应受到所用涂层类型的显著调节。
