文章摘要
盛燕辉,李世杰,孔祥清,杨 荣,陈 洁,李胜男,曹克将.体外观察壳聚糖/重组水蛭素共价包被先天性心脏病介入封堵材料的生物相容性[J].南京医科大学学报,2008,28(12):1527~1532
体外观察壳聚糖/重组水蛭素共价包被先天性心脏病介入封堵材料的生物相容性
Biocompatibility of Nitinol slices designed for the closure of CHD covalently coated by chitosan/recombinant-hirudin in vitro
投稿时间:2008-08-11  
DOI:10.7655
中文关键词: 壳聚糖/重组水蛭素  镍钛合金  共价包被  生物相容性
英文关键词: Chitosan/r-hirudin  Nitinol  covalently coated  biocompatibility
基金项目:江苏省"135"重点工程课题资助项目(039);江苏省科委社会发展基金课题(BS2002053)
作者单位
盛燕辉 南京医科大学第一附属医院心血管内科,江苏 南京 210029 
李世杰 南京医科大学第一附属医院心血管内科,江苏 南京 210029 
孔祥清 南京医科大学第一附属医院心血管内科,江苏 南京 210029 
杨 荣 南京医科大学第一附属医院心血管内科,江苏 南京 210029 
陈 洁 南京医科大学心血管药理研究室,江苏 南京 210029 
李胜男 南京医科大学心血管药理研究室,江苏 南京 210029 
曹克将 南京医科大学第一附属医院心血管内科,江苏 南京 210029 
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中文摘要:
      目的:体外评价壳聚糖/重组水蛭素(Chi/r-Hir)共价包被先天性心脏病介入封堵材料镍钛合金的生物相容性?方法:①实验分6组:对照组(未接触镍钛合金的血液)?未包被组?Chi组和Chi/r-Hir低浓度?中浓度和高浓度组;②血液相容性实验:包括血液溶血试验和动态全人血接触试验?观察实验各组镍钛合金片溶解红细胞的特性;检测动态全人血接触后实验各组血液APTT?PT和TT,观察其抗血栓性能;扫描电子显微镜观察动态全人血接触后实验各组镍钛合金片表面血小板黏附?聚集和纤维蛋白吸附情况?③细胞相容性实验:将体外培养人脐静脉内皮细胞株(HUVECs)种植于实验各组镍钛合金表面,孵育72 h,分别应用Fn和Ki67免疫荧光抗体标记,在荧光显微镜下观察HUVECs在镍钛合金表面的黏附?增殖情况并进行半定量分析?结果:①实验各组溶血率均小于1.7%;②动态全人血接触后,实验各组红细胞?白细胞?血小板计数均在正常值范围内且组间未见差异;与未包被组和Chi组相比,Chi/r-Hir包被明显延长APTT和TT(P < 0.001);Chi/r-Hir包被延长APTT和TT作用与重组水蛭素浓度有关,低?中?高浓度组间相比存在差异(方差不齐采用Dunnetts法P < 0.05);③扫描电镜结果显示,Chi/r-Hir组与Chi组相比,镍钛合金表面血小板黏附?聚集和纤维蛋白吸附明显减少?④细胞相容性实验:经过72 h孵育,镍钛合金片边缘细胞生长和移行良好?Fn荧光标记观察HUVECs在镍钛合金片表面黏附依次为:Chi组>未包被组>Chi/r-Hir组,Chi/r-Hir包被不同浓度组间HUVECs黏附未见差异;Ki67免疫荧光标记观察HUVECs在镍钛合金表面增殖依次为:未包被组 = Chi组 > Chi/r-Hir组,Chi/r-Hir包被不同浓度组间HUVECs增殖未见差异?结论:Chi/r-Hir包被先天性心脏病介入封堵材料镍钛合金后,具有良好抗凝性能,但不利于HUVECs黏附和增殖?
英文摘要:
      Objective:To assess the effect of bioactive coatings on biocompatibility of flat Nitinol slices designed for the closure of congenital heart defects in vitro. Methods:①The experiment was divided into six groups: control group, uncoating group, chitosan group(Chi group), chitosan/r-hirudin(Chi/r-Hir) low-dose group, Chi/r-Hir middle-dose group and Chi/r-Hir high-dose group. ②The hemocompatibility of biomaterials was evaluated in vitro in a dynamic model with whole human blood by the hemolysis test and anticoagulant activity such as partial thrombin time(APTT), prothrombin time(PT) and thrombin time(TT) were measured as well. Surface characterizations such as protein adsorption, platelet adhesion and aggregation were observed by scanning electron microscopy. ③Humanumbilical vein endothelial cells(HUVECs) were cultured in the presence of tested slices for 72 h. Adhesion and proliferation of HUVECs were evaluated with light microscopy and fluorescence microscope by double-staining of fibronectin(Fn) and Ki67. Results:①The hemolysis rates of biomaterials were less than 1.7%. ②White blood count(WBC), red blood count(RBC) and platelet count (PC) were normal and no difference was observered between each groups. ③Chi/r-Hir significantly prolong the APTT and TT than those of other groups(P < 0.001). the protein adsorption, platelet adhesion and aggregation were reduced by Chi/r-Hir coated. furthermore, there was dose-effect relationship of differential concentration Chi/r-Hir coated(Dunnetts, P < 0.05). ④After 72 h of incubation, without abnormal morphology, HUVECs were able to grow on to all kinds of slices. The adhesion of the cells on the slices by fluorescence microscopy through the application of Fn immunofluorescent antibody labeled HUMECs decreased as following:Chi>Uncoating>Chi/r-Hir coating. The proliferation of the cells on the slices by fluorescence microscopy through the application of Ki67 immunofluorescent antibody labeled HUMECs decreased as following:Chi=uncoating>Chi/r-hir coating. The cells grown on the surfaces of Chi/r-Hir were not as good as on the Nitinols slices. Conclusion:In vitro, Chi/r-Hir coated with nitinol slices present good anti-coagulation, but adhesion and proliferation of HUVEC is not beneficial.
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