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基于聚(羟基丁酸-co-羟基戊酸酯)的工程血管支架,以模拟天然动脉。

Engineering poly(hydroxy butyrate-co-hydroxy valerate) based vascular scaffolds to mimic native artery.

机构信息

Centre for Nanosciences and Molecular Medicine, Amrita Institute of Medical Sciences and Research Centre, Amrita Vishwa Vidyapeetham, Kochi 682041, India.

Centre for Nanosciences and Molecular Medicine, Amrita Institute of Medical Sciences and Research Centre, Amrita Vishwa Vidyapeetham, Kochi 682041, India.

出版信息

Int J Biol Macromol. 2018 Apr 1;109:85-98. doi: 10.1016/j.ijbiomac.2017.12.077. Epub 2017 Dec 13.

Abstract

Electrospun tri-layered fibrous scaffold incorporating VEGF and Platelet Factor Concentrate (PFC) in multiple layers having different layer architectures was designed to mimic native artery. The scaffold consisted of longitudinally aligned poly(hydroxy butyrate-co-hydroxy valerate) (PHBV) and poly(vinyl alcohol) (PVA) nanofibers (inner layer), radially aligned PHBV-elastin nanofibers (middle layer) to provide the bi-directional alignment and combination of longitudinally aligned PHBV-elastin and random PHBV/PVA multiscale fibers (peripheral layer). Tubular constructs of diameter <6 mm were developed. The developed electrospun fibers were characterised by Scanning Electron Microscope (SEM), Fourier Transform Infrared Spectroscopy and Tensile tests. Further the burst strength, compliance and stiffness index of tri-layered tubular scaffold was evaluated. SEM images of fibrous layers showed the typical longitudinal and radial alignment of fibers in the tubular construct. SEM images showed that the prepared PHBV nanofibers were in the range of 500-800 nm and PHBV microfibers were of 1-2 μm in diameter in the tri-layered electrospun membrane. PVA nanofibers were of size 200-250 nm. The tensile strength, percentage compliance and stiffness index of tri-layered membrane was in accordance with that of native small blood vessels. The developed tri-layered membrane was blood compatible, with hemolysis degree 0.85 ± 0.21% and did not activate platelets. Controlled release of VEGF and PFC was observed from the scaffold. The biocompatibility of the tri-layered scaffold was evaluated using HUVECs, SMCs and MSCs and SMCs infiltration from the outer layer was also evaluated. Specific protein expression for the HUVECs and SMCs was evaluated by flow cytometry and immunocytochemistry. HUVECs and SMCs exhibited good elongation and alignment along the direction of fibers and was found to maintain its CD31, VE-Cadherin and αSMA expression respectively. The results highlight the importance of bi-directional fiber alignment on the tri-layered electrospun scaffold as a suitable architectural prototype for vascular scaffolds to mimic the native arteries.

摘要

设计了一种具有多层不同层结构的电纺三层纤维支架,其中包含血管内皮生长因子(VEGF)和血小板因子浓缩物(PFC),以模拟天然动脉。支架由纵向排列的聚(羟基丁酸-co-羟基戊酸)(PHBV)和聚乙烯醇(PVA)纳米纤维(内层)组成,径向排列的 PHBV-弹性蛋白纳米纤维(中层)提供双向排列和纵向排列的 PHBV-弹性蛋白与随机 PHBV/PVA 多尺度纤维(外围层)的组合。开发了直径<6mm 的管状结构。通过扫描电子显微镜(SEM)、傅里叶变换红外光谱和拉伸试验对所开发的电纺纤维进行了表征。进一步评估了三层管状支架的爆裂强度、顺应性和刚度指数。纤维层的 SEM 图像显示了管状结构中纤维的典型纵向和径向排列。SEM 图像显示,制备的 PHBV 纳米纤维的直径在 500-800nm 范围内,PHBV 微纤维的直径在 1-2μm 范围内,三层电纺膜中的 PVA 纳米纤维的尺寸为 200-250nm。三层膜的拉伸强度、百分比较大顺应性和刚度指数与天然小血管一致。所开发的三层膜具有血液相容性,溶血度为 0.85±0.21%,不会激活血小板。从支架中观察到 VEGF 和 PFC 的控制释放。通过 HUVECs、SMC 和 MSC 评估了三层支架的生物相容性,并评估了外层 SMC 的渗透情况。通过流式细胞术和免疫细胞化学评估了 HUVECs 和 SMCs 的特定蛋白表达。HUVECs 和 SMCs 沿纤维方向表现出良好的伸长和排列,并发现其分别保持 CD31、VE-Cadherin 和 αSMA 的表达。结果强调了三层电纺支架中纤维双向排列的重要性,作为一种适合的血管支架结构原型,可模拟天然动脉。

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