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通过立即接种源自网膜组织的自体内皮细胞,犬体内涤纶下腔静脉假体的快速细胞腔内覆盖:一项初步试验的结果

Rapid cellular luminal coverage of Dacron inferior vena cava prostheses in dogs by immediate seeding of autogenous endothelial cells derived from omental tissue: results of a preliminary trial.

作者信息

Wang Z G, Du W, Li G D, Pu L Q, Sharefkin J B

机构信息

Department of Vascular Surgery, Beijing Heart, Lung, and Blood Vessel Medical Center, China.

出版信息

J Vasc Surg. 1990 Aug;12(2):168-79.

PMID:2143236
Abstract

Endothelial cell seeding methods might reduce the high failure rates of venous vascular prostheses, but low flow rates in venous vascular prostheses impose a need to protect early patency and to attain early endothelial cell coverage without waiting several weeks for relatively small endothelial cell innocula from autologous veins to form confluent linings. To obtain large number of autologous endothelial cells for high-density seeding, canine omental microvascular endothelial cells were harvested by collagenase digestion and density gradient centrifugation, with yields of 1.34 +/- 0.24 (SD) X 10(6) cells/gm of omentum (N = 8 harvests). Primary culture of a subfraction from each harvest showed the cell population to be dominated by factor VIII-related antigen-positive endothelial cells with only a few nonstaining cells (estimated to be 10% or less of total cell number) visible. Freshly harvested omental cells were seeded onto double velour knitted Dacron prostheses at densities of 5 X 10(5) cells/cm2 of graft luminal surface in an autologous plasma suspension by use of prior preclotting with cell-free autologous plasma, followed by endothelial cell seeding in autologous plasma, with plasma recalcification during endothelial cell instillation. Six seeded and two control (sham-seeded) vascular prostheses 5 cm long with 10 mm inner diameter were used as inferior vena cava interposition grafts. A distal arteriovenous fistula and aspirin (300 mg) and dipyridamole (50 mg orally every day) starting 3 days before surgery were used to protect early patency of all grafts. Seeded venous vascular prostheses were explanted for study at intervals of 1,5, and 10 days after surgery (N = 2 prostheses at each time); the two control venous vascular prostheses were explanted at 10 days. All venous vascular prostheses were patent at time of removal. In seeded venous vascular prostheses, light, scanning, and transmission electron microscopy showed emergence of numerous flattened endothelial cell-like cells on the luminal surface 24 hours after surgery, followed by formation of a confluent cellular lining without adherent platelets by 5 to 10 days after surgery. Control venous vascular prostheses, in contrast, remained covered by an irregularly thickened fibrin and red cell thrombus, which sometimes encroached on the lumen. Our results suggest that (1) omental tissue can furnish endothelial cells for high-density immediate seeding of venous vascular prostheses, and (2) that the method we used to combine features of both so-called high density "seeding" and "sodding" techniques offers both more rapid prosthesis coverage than the former and shorter intraoperative times for cell attachment to prostheses than the latter.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

内皮细胞接种方法可能会降低静脉血管移植物的高失败率,但静脉血管移植物中的低流速使得有必要保护早期通畅性,并在无需等待数周让来自自体静脉的相对少量内皮细胞接种物形成融合内膜的情况下实现早期内皮细胞覆盖。为了获得大量自体内皮细胞用于高密度接种,通过胶原酶消化和密度梯度离心收获犬网膜微血管内皮细胞,每克网膜的产量为1.34±0.24(标准差)×10⁶个细胞(n = 8次收获)。每次收获的一个亚组分的原代培养显示细胞群体以因子VIII相关抗原阳性的内皮细胞为主,仅可见少数不着色细胞(估计占细胞总数的10%或更少)。将新鲜收获的网膜细胞以5×10⁵个细胞/cm²的移植物腔表面密度接种到双层丝绒编织涤纶移植物上,接种于自体血浆悬液中,先用无细胞自体血浆预凝,然后在内皮细胞接种时进行血浆再钙化。使用6个接种的和2个对照(假接种)的内径为10 mm、长5 cm的血管移植物作为下腔静脉间置移植物。在手术前3天开始建立远端动静脉瘘,并给予阿司匹林(300 mg)和双嘧达莫(每天口服50 mg)以保护所有移植物的早期通畅性。接种的静脉血管移植物在手术后1天、5天和10天间隔取出进行研究(每次时间点n = 2个移植物);2个对照静脉血管移植物在10天时取出。所有静脉血管移植物在取出时均通畅。在接种的静脉血管移植物中,光镜、扫描电镜和透射电镜显示术后24小时在腔表面出现大量扁平的内皮细胞样细胞,术后5至10天形成无黏附血小板的融合细胞内膜。相比之下,对照静脉血管移植物仍被不规则增厚的纤维蛋白和红细胞血栓覆盖,有时会侵入管腔。我们的结果表明:(1)网膜组织可为静脉血管移植物的高密度即时接种提供内皮细胞;(2)我们用于结合所谓高密度“接种”和“铺草皮”技术特点的方法,比前者提供更快的移植物覆盖,比后者细胞附着于移植物的术中时间更短。(摘要截短至400字)

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