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带孔聚四氟乙烯移植物的再内皮化:脉冲灌注条件下的体外研究

Re-endothelialization of punctured ePTFE graft: an in vitro study under pulsed perfusion conditions.

作者信息

Mall Julian W, Philipp Andreas W, Rademacher Axel, Paulitschke Manrico, Büttemeyer Rolf

机构信息

Department of General, Visceral, Thoracic and Vascular Surgery, Medical Faculty of the Humboldt University, Charitè, Campus Mitte, Schumannstrasse 20/21, D-10117 Berlin, Germany.

出版信息

Nephrol Dial Transplant. 2004 Jan;19(1):61-7. doi: 10.1093/ndt/gfg319.

Abstract

BACKGROUND

When used as arteriovenous (AV) shunts for haemodialysis, small diameter expanded polytetrafluoroethylene (ePTFE) grafts have a high failure rate in vivo. Attempts to improve graft patency are various, and focus on either improvement of implantation techniques or graft tissue engineering. The tissue engineering approach attempts to reproduce in grafts the properties of pristine vasculature. As shown in previous experiments, it is possible to grow on ePTFE grafts under shear stress in vitro an autologous endothelial cell layer, which will withstand physiological stress under in vivo conditions of blood flow. The aim of this study was to investigate in an in vitro model the regenerative potency of a tissue-engineered prosthetic vascular graft after repeated cannulation with a haemodialysis cannula.

METHODS

Pig endothelial cells were harvested from an external jugular vein. Following processing of the endothelial cells, seven ePTFE grafts were coated with an inner cell layer and were kept under pulsed perfusion. Each graft was then cannulated three times with a standard shunt needle. The endothelium was then left to regenerate for a maximum of 48 h. The grafts were stained with haematoxylin/eosin before histological study.

RESULTS

All grafts were endothelialized over the puncture sites within 48 h. Histological analysis revealed a confluent endothelial cell lining at each puncture site. Cell morphology and cell pattern over puncture sites were not different from randomly picked locations over the graft lumen.

CONCLUSION

Our results underline the potential of endothelial tissue engineering in vascular shunt surgery. Vascular bio-hybrids that have the properties of pristine vascular endothelium may be a key step forward in maintaining angio-access in patients who require haemodialysis.

摘要

背景

小直径的膨体聚四氟乙烯(ePTFE)移植物在用作血液透析的动静脉(AV)分流器时,在体内具有较高的失败率。改善移植物通畅性的尝试多种多样,主要集中在改进植入技术或移植物组织工程方面。组织工程方法试图在移植物中重现原始脉管系统的特性。如先前实验所示,在体外剪切应力作用下,有可能在ePTFE移植物上生长出自体内皮细胞层,该细胞层在体内血流条件下能够承受生理压力。本研究的目的是在体外模型中研究经血液透析插管反复穿刺后组织工程化人工血管移植物的再生能力。

方法

从猪颈外静脉采集内皮细胞。在内皮细胞处理后,将七个ePTFE移植物涂上内层细胞层,并保持脉冲灌注。然后用标准分流针将每个移植物穿刺三次。之后让内皮细胞再生最多48小时。在进行组织学研究之前,用苏木精/伊红对移植物进行染色。

结果

所有移植物在48小时内穿刺部位均实现内皮化。组织学分析显示每个穿刺部位有融合的内皮细胞内衬。穿刺部位的细胞形态和细胞模式与移植物管腔内随机选取的部位没有差异。

结论

我们的结果强调了内皮组织工程在血管分流手术中的潜力。具有原始血管内皮特性的血管生物杂交体可能是维持需要血液透析患者血管通路的关键一步。

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