Flow shear stress promotes the endothelialization of small diameter tissue-engineered vessels

doi:10.3969/j.issn.2095-4344.2017.16.010

Abstract

Abstract:

BACKGROUND: Magnitude and action ways of flow shear stress are important for the endothelialization of small diameter tissue-engineered vessels (TEV), and the TEV resistance ability to blood flow even decides the destiny of implantation. There are many studies on how to construct the TEV and improve anticoagulant ability of TEV after host cell implantation, but the effects of different flow shear stresses on the TEV endothelialization is rarely reported, which may be helpful for increasing the success rate of TEV implantation.
OBJECTIVE: To compare the effects of flow shear stresses in single level or stepwise increased on the endothelialization of small diameter TEV and to optimize the TEV in the aspects of shear stress magnitude and loading method.
METHODS: The number, morphology and adhesion ability of endothelial cells on the inner wall of TEV were observed through silver nitrate and F-actin staining.
RESULTS AND CONCLUSION: Single-level shear stress at 2.5, 3.0 N/m² for 2 hours removed almost all the endothelial cells seeded on the inner wall of TEV. In contrast, stepwise increased shear stress from 0.5 N/m² to 3.0 N/m2 at an increase of 0.2 N/m²/2 hours maintained the integrity and oriented along the flow direction, and could induce stress fibers productionin endothelial cells. These results suggest that the stepwise increased flow shear stress can improve the endothelialization of TEV.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Endothelial Cells, Biomechanics, Tissue Engineering

CLC Number:

R318

Zhang Wei, Song Ai-jing. Flow shear stress promotes the endothelialization of small diameter tissue-engineered vessels [J]. Chinese Journal of Tissue Engineering Research, 2017, 21(16): 2515-2520.

Figures/Tables 1

2.1 大鼠颈总动脉的形态硝酸银染色法和F-actin荧光标记染色结果显示，大鼠颈动脉血管内壁上无血管内皮细胞，但血管结构保留完整(图2)。 2.2 脱细胞大鼠颈总动脉的内皮化硝酸银染色和血管内皮细胞 F-actin荧光标记结果表明，血管内皮细胞已完全覆盖血管内壁表面，生长状态良好，并融合成细胞单层；细胞呈椭圆形，细胞间连接紧密；细胞膜周边出现F-actin(图3)。 2.3 流动剪切力对组织工程血管内皮化的作用 2.3.1 单一水平的流动剪切力硝酸银染色结果表明，0.5 N/m2作用24 h，细胞无明显变化；1.0 N/m2作用24 h，部分细胞变得紧缩，有少许细胞脱落；1.5，2.0 N/m2作用2 h，部分细胞呈梭形，局部有细胞脱落；2.5 N/m2作用2 h，细胞形态呈明显的梭形，细胞的长轴与液体流动方向一致，细胞脱落较多，细胞膜受到不同程度的损伤；3.0 N/m2作用2 h，细胞几乎全部脱落，仅存的个别细胞形态为椭圆形，可清晰看到血管壁内弹性层(图4)。荧光染色结果显示，0.5 N/m2作用24 h，微丝主要分布在细胞膜的周边；1.0 N/m2作用24 h，细胞膜周边微丝减少，核周边微丝数量增多；1.5，2.0 N/m2作用2 h，核周微丝逐渐增多；2.5 N/m2作用2 h，微丝主要分布在核周边，能清晰看到应力纤维；3.0 N/m2作用2 h，细胞几乎全部脱落，观察不到微丝的存在(图4)。 2.3.2 成梯度增加的流动剪切力硝酸银染色结果表明，经0.5-1.5 N/m2成梯度增加的剪切力作用后，多数细胞保持原状，极少细胞脱落；0.5-2.0 N/m2成梯度增加的剪切力作用后，部分细胞变为梭形，细胞长轴与液流方向一致；细胞间连接紧密，无细胞脱落；0.5-2.5 N/m2成梯度增加的剪切力作用后，多数细胞变为梭形，有明显的轴向排列；细胞间连接紧密，极少数细胞脱落；0.5-3.0 N/m2成梯度作用后，细胞呈梭形，沿液流方向被拉长，有明显的轴向排列，内皮细胞层较为完整(图5)。荧光染色结果显示，经0.5-1.5 N/m2成梯度增加的剪切力作用后，微丝主要分布在膜周边；0.5-2.0 N/m2成梯度增加的剪切力作用后，部分细胞变为梭形，细胞长轴与液流方向一致，细胞间连接紧密；核周边微丝明显增多；0.5-2.5 N/m2成梯度增加的剪切力作用后，细胞多为梭形，有明显的轴向排列；微丝主要集中在核周边，有应力纤维的出现；0.5-3.0 N/m2成梯度作用后，细胞有明显的轴向排列，微丝主要分布在核周边，有应力纤维的出现(图5)。"

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