纤维蛋白组织粘着剂携载反义基因片段的实验研究

EXPERIMENTAL STUDY ON FIBRIN TISSUE ADHESIVE CARRYING ANTISENSE OLIGODEOXYNUCLEOTIDE

为了探讨纤维蛋白组织粘着剂携载反义基因片段血管外膜给药的可能性及可行性 ,作者首先对粘着剂的物理性状进行了观察 ,继而将亚甲蓝溶于粘着剂中观察其上清液的颜色变化 ,再以人工合成的反义寡核苷酸溶于粘着剂中 ,分别于 12、2 4、36、48、6 0、72h取其上清液以紫外分光光度计测其OD值 ,绘出其体外控释曲线。结果发现 ,粘着剂混合后即刻凝结 ,表面呈膜状 ,内部呈丝状蛋白结构 ,其间充满颗粒状液体 ;粘着剂具有弹性 ,有类似海绵样特性 ,且随着时间延长其上清液颜色渐变深 ;体外控释曲线显示其携载的反义寡核苷酸可以缓慢释放 ,近 72h已大部分释放。提示纤维蛋白组织粘着剂可以携载反义基因片段直接血管外膜给药 ,使反义药物缓慢释放 。

In order to explore the possibility and feasibility of administering antisense oligodeoxynucleotide to vascular adventia delivering by fibrin tissue adhesive, the authors first observed the physical characteristics of the adhesive, then the blue dye (methylenophin) was dissolved in the adhesive and the color change of the supernate was monitored. The adhesive with antisense oligomer inside was immerged in saline and the optical density (OD) value of the released oligomer was measured by ultraviolet spectrophotometer at different time points, then the in vitro controlled releasing curve was made. The preliminary in vitro test found the adhesive clot in 10 seconds when the two kinds of solution were mixed together, and the surface was membrane like under microscope, the clot inside was silk protein like configuration and was filled up with particle like liquid. The clot was elastic and had some reabsorbable property like that of sponge. The blue dye inside the clot gave a more and more significant dyeing of the supernate with time going. The in vitro releasing curve indicated that most oligomer was released within 72 hours. This implied that the adhesive clot is like a sponge reservoir of drugs, it could elongate the action time of antisense drug at vascular adventia.