Patterson R B, Mayfield G, Silberstein E B, Kempczinski R F
Department of Surgery, University of Cincinnati Medical Center, OH 45267-0558.
J Vasc Surg. 1989 Dec;10(6):650-5. doi: 10.1067/mva.1989.15992.
The current widespread use of indium 111 oxine for labeling endothelial cells to study their interaction with various bioprosthetic flow surfaces presupposes a high retention of the radioisotope within the cell and a lack of significant adherence of any marker released from the cell to the surface under study. We measured the loss of indium 111 from freshly harvested, canine, venous endothelial cells, and their viability in cell culture, for varying intervals up to 24 hours. At prescribed intervals, aliquots of the radiolabeled endothelial cell suspension were centrifuged, and the supernatant was separated from the cell pellet. The relative radioactivity of each was measured in a gamma well counter and the spontaneous loss of marker from the endothelial cells was calculated. Spontaneous loss of indium 111 was 7.8% +/- 1.9% at 1 hour and 47.6% +/- 1.8% at 24 hours. Loss of activity was virtually constant between 14 and 24 hours. Endothelial cell viability was 80% at 24 hours. We next studied the in vitro affinity of indium 111 oxine and indium 111 transferrin for untreated expanded polytetrafluoroethylene vascular grafts and for similar grafts treated with two surfactants commonly used to increase the "wetability" of expanded polytetrafluoroethylene, a nonionic sufectant (Nonidet) and tridodecylmethylammonium chloride, and with two glycoproteins, fibronectin and basement membrane gel. A minimum of three graft segments were studied in each group. The affinity of graft material for indium 111, in both its oxine and transferrin complex, was significantly increased by treating the graft with a wetting agent, and it was further increased by the addition of a glycoprotein.(ABSTRACT TRUNCATED AT 250 WORDS)
目前广泛使用铟 - 111 奥克辛(铟 - 111 与8 - 羟基喹啉的络合物)标记内皮细胞以研究其与各种生物假体血流表面的相互作用,这预先假定放射性同位素在细胞内有高保留率,并且从细胞释放的任何标记物不会显著附着在所研究的表面上。我们测量了新鲜采集的犬静脉内皮细胞在长达24小时的不同时间段内铟 - 111的损失情况以及它们在细胞培养中的活力。在规定的时间间隔,将放射性标记的内皮细胞悬液的等分试样进行离心,使上清液与细胞沉淀分离。在伽马孔计数器中测量各自的相对放射性,并计算内皮细胞中标记物的自发损失。铟 - 111的自发损失在1小时时为7.8%±1.9%,在24小时时为47.6%±1.8%。在14至24小时之间活性损失基本恒定。24小时时内皮细胞活力为80%。接下来,我们研究了铟 - 111奥克辛和铟 - 111转铁蛋白对未处理的膨体聚四氟乙烯血管移植物以及用两种常用于增加膨体聚四氟乙烯“润湿性”的表面活性剂(一种非离子表面活性剂(诺乃洗涤剂)和氯化三(十二烷基)甲基铵)以及两种糖蛋白(纤连蛋白和基底膜凝胶)处理的类似移植物的体外亲和力。每组至少研究三个移植物片段。通过用湿润剂处理移植物,其对铟 - 111(以其奥克辛和转铁蛋白络合物形式)的亲和力显著增加,并且通过添加糖蛋白进一步增加。(摘要截短于250字)