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Decellularized ovine arteries as small-diameter vascular grafts

机译:脱细胞的绵羊动脉作为小直径血管移植物

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Atherosclerosis and its complications still represent the leading cause of death in the developed countries. While autologous blood vessels may be regarded as the best solution for peripheral and coronary bypass, they are unavailable in most patients. Even though tissue engineering techniques are often applied to the development of small-diameter vascular grafts, limiting factors of this approach are represented by the lack of essential extracellular matrix proteins and/or poor biomechanical properties of the scaffolds used. Along these lines, the aim of this study was to develop a decellularization protocol for ovine carotids to be used as suitable small-diameter vascular grafts. Samples were treated either with sodium dodecyl sulphate (SDS) or with Trypsin and Triton X-100; a final nuclease digestion was performed for both protocols. Morphological analyses demonstrate complete removal of nuclei and cellular components in treated vessels, also confirmed by significant reduction in wall thickness and DNA content. Essential extracellular matrix proteins such as collagen, elastin, and fibronectin are well preserved after decellularization. From a mechanical point of view, Trypsin and Triton X-100 treated arteries show elastic modules and compliance comparable to native carotids, whereas the use of SDS makes samples stiffer, with a significant decrease in the compliance mean value and an increase in longitudinal and circumferential Young's modules. It is demonstrated that the treatment where Trypsin and Triton X-100 are combined guarantees complete decellularization of carotids, with no significant alteration of biomechanical and structural properties, thus preserving a suitable environment for adhesion, proliferation, and migration of cells.
机译:动脉粥样硬化及其并发症仍然是发达国家的主要死亡原因。自体血管可能被认为是外周和冠状动脉搭桥术的最佳解决方案,但大多数患者无法使用。即使组织工程技术经常用于开发小直径血管移植物,但这种方法的局限性因素仍然是缺乏必需的细胞外基质蛋白和/或所用支架的生物力学性能较差。沿着这些思路,本研究的目的是为羊颈动脉制定脱细胞协议,以用作合适的小直径血管移植物。样品用十二烷基硫酸钠(SDS)或胰蛋白酶和Triton X-100处理。两种方案均进行了最终的核酸酶消化。形态分析表明,处理过的血管中细胞核和细胞成分已完全清除,这也通过壁厚和DNA含量的显着降低得到证实。脱细胞后,基本的细胞外基质蛋白(例如胶原蛋白,弹性蛋白和纤连蛋白)可以很好地保存。从机械角度来看,胰蛋白酶和Triton X-100处理过的动脉显示出与天然颈动脉相当的弹性模量和顺应性,而SDS的使用使样品更坚硬,顺应性平均值显着下降,纵向和周向增大杨氏模块。已经证明,胰蛋白酶和Triton X-100结合使用的治疗方法可确保颈动脉完全脱细胞,而不会显着改变生物力学和结构性质,从而为细胞的粘附,增殖和迁移保留了合适的环境。

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