Cardiac and lung endothelial cells in response to fluid shear stress on physiological matrix stiffness and composition

Abstract

Objective 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. Methods A novel polydimethylsiloxane‐based parallel plate flow chamber enabled application of laminar fluid shear stress of 2 dynes/cm2 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. Results 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. αv and β3 integrin expression decreased after application of shear stress in both cell types. Conclusion 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.

Organ-Specific Endothelial Cell Differentiation and Impact of Microenvironmental Cues on Endothelial Heterogeneity

“…In all cases, researchers must decide what source of endothelial is the most suitable. As can be observed in Table 1, many of them use commercially available organ-specific endothelial cells [118][119][120][121][122][123][124][125][126][127][128] and only a few freshly isolate the endothelial cells they work with [106,[129][130][131][132]. Others use human embryonic or pluripotent stem cells (hESCs or hPSCs), co-/differentiated into endothelial cells in the presence (or not) of other stromal cells (such as pericytes or smooth muscle cells) or organotypic parenchymal cells [133][134][135].…”

“…Others use human embryonic or pluripotent stem cells (hESCs or hPSCs), co-/differentiated into endothelial cells in the presence (or not) of other stromal cells (such as pericytes or smooth muscle cells) or organotypic parenchymal cells [133][134][135]. Another option is to genetically modify organ-specific endothelial cells after isolation to force them to maintain their phenotypical cues for the longer term [124,125,136]. Yet, many researchers still chose insufficiently specialized endothelial cells, such as human umbilical vein endothelial cells (HUVECs) [137][138][139][140].…”

“…In an attempt to neutralize this loss, some groups have investigated the use of genetic modifications like an adenoviral genetic modification (transfection of E4ORF1) in primary endothelial cells (from lung and heart) to avoid cellular transformation along passages, preserving endothelial features in a longer-term [124,125]. Similarly, Palikuqi et al transiently overexpressed the embryonic ETV2 factor in mature human endothelial cells in vitro (detailed in Table 1) and co-cultured these with organoid models (e.g., pancreatic islets), obtaining organ-specific endothelial cell cues [136].…”

“…Similarly, Palikuqi et al transiently overexpressed the embryonic ETV2 factor in mature human endothelial cells in vitro (detailed in Table 1) and co-cultured these with organoid models (e.g., pancreatic islets), obtaining organ-specific endothelial cell cues [136]. However, the authors warn that their results may differ from those obtained by native endothelial cell counterparts [124,125]. The use of low specialized endothelial cells has become, therefore, one of the most suitable solutions to ensure a greater reproducibility in vascular research, even if such results have compromised specificity.…”

In Vitro Measurements of Cellular Forces and their Importance in the Lung—From the Sub- to the Multicellular Scale

“…To investigate the cellular mechanics of cells and tissue that are under constant shear stress, as in blood vessels or lungs, microfluidic devices [ 159 , 160 , 161 , 162 ] and mechanical stretchers [ 163 , 164 , 165 , 166 ] can be used. In these devices, the cell usually experiences a shear stress, resulting from the strained (stretched) substrate or the moving fluid.…”