磺酸根离子化及白蛋白预吸附对胆管支架细菌吸附影响的体外研究
发布时间：2018-10-15 16:52
【摘要】：背景梗阻性黄疸是由胆结石或肝胆部位的肿瘤所造成的黄疸,在众多的治疗方法中,介入人工胆管支架是一种有效的治疗方法；商业化聚氨基甲酸酯材质的人工胆管也在临床上应用,且此材质在胆汁的测试中有不错的成效,然而胆汁中生物膜的形成以及沉积物的附着导致人工胆管支架再阻塞,从而缩短了使用期限,因此,改变胆管支架材质的表面性质是防止再堵塞的有效方法。目的通过对聚氨基甲酸酯作磺酸根离子基团的改质,同时以白蛋白吸附进行表面处理,探讨对胆管支架表面细菌粘附、胆汁沉积物和胆管支架堵塞的影响。方法1. 将氢化钠(NaH)加入PU溶液进行双分子亲核取代反应,之后再加上丙烷硫酸酯进行开环反应,最后产生不同磺酸根离子含量的PU,涂层于聚乙烯(PE)及氧化聚乙烯管(OxPE)2.选择白蛋白浓度1mg/ml,预吸附时间10分钟与60分钟,将材质为聚乙烯、氧化聚乙烯、磺酸化聚氨酯、基聚氨酯的支架经紫外线照射,以串联方式连接,两端接三通管,通入缓冲液并在管内停留12小时以上3. 针对不同磺酸根离子含量及蛋白质吸附10分钟及60分钟的聚氨基甲酸酯,在浓度为4-9×107 CFU/mL DH5 a型大肠杆菌的人体胆汁及PBS中做体外24小时连续循环实验。4. 实验经24小时后,将支架以PBS润洗,流体入口的前3厘米舍弃,之后每1厘米取样一次,共取三次进行表面吸附细菌的定量,另外再取1厘米,使用含2.5%戊二醛的PBS溶液进行表面细菌的固定化6小时,并进行干燥处理程序。接着材质进行镀金处理,以扫描式电子显微镜(SEM)进行表面细菌吸附情形的观察。结果1. SEM图可观察到在胆汁循环实验方面,高磺酸根离子含量的PU能减少细菌及沉积物的吸附；在缓冲液循环实验方面,没有沉积物的生成,且表面吸附的细菌没有聚集的现象,大多呈单层吸附,并平均分散在材质的表面。在细菌吸附量分析方面,显示随着磺酸基含量的增加,表面细菌的吸附量逐渐减少。2. 材质经白蛋白的预吸附处理后,在缓冲液系统下,发现经白蛋白预吸附后的材质表面有较少的细菌吸附,且随白蛋白预吸附时间增加,细菌吸附量有减少的趋势。3. PE、OxPE、Pell-S-0.10经白蛋白预吸附处理后,抑制细菌吸附的效果好；且在P E、OxPE中,白蛋白预吸附60分钟比10分钟其细菌吸附量更少,其差异具有统计学意义(P0.05),在Pell-S-0.10中,白蛋白预吸附60分钟与10分钟细菌吸附量无明显差异。结论本实验显示聚氨基甲酸酯磺酸根离子化及白蛋白预吸附能减少胆管支架表面细菌生物膜的形成以及胆汁沉积物的附着,对减少胆管支架再阻塞可能有一定价值。
[Abstract]:Background obstructive jaundice is a kind of jaundice caused by gallstone or hepatobiliary tumor. Among many treatment methods, interventional artificial bile duct stent is an effective treatment method. Commercial polycarbamate artificial bile ducts are also used in clinical practice, and this material has good results in bile testing. However, the formation of biofilm in bile and the adhesion of sediments lead to re-obstruction of artificial bile duct stents. Therefore, changing the surface properties of bile duct scaffolds is an effective way to prevent re-clogging. Objective to study the effect of polycarbamate on bacterial adhesion, bile sediment and obstruction of bile duct scaffolds by surface modification of sulfonic groups and albumin adsorption. Method 1. Sodium (NaH) was added to PU solution to undergo bimolecular nucleophilic substitution reaction, followed by ring opening reaction with propane sulfate. Finally, PU, coating with different sulfonic ion content was produced in polyethylene (PE) and (OxPE) 2 oxide polyethylene tube. Select albumin concentration 1 mg / ml, preadsorption time 10 minutes and 60 minutes, the material of polyethylene, oxide polyethylene, sulfonated polyurethane, UV irradiation, connected in series, the two ends of the tube, Enter the buffer and stay in the tube for more than 12 hours. Aiming at different sulfonic ion content and protein adsorption for 10 minutes and 60 minutes of polycarbamate, the human bile and PBS containing 4-9 脳 107 CFU/mL DH5 a Escherichia coli were subjected to continuous cardiopulmonary bypass for 24 hours. 4. After 24 hours, the scaffold was washed with PBS, the first 3 cm of fluid inlet was discarded, then every 1 cm was sampled. The surface bacteria were immobilized in PBS solution containing 2.5% glutaraldehyde for 6 hours, and the drying process was carried out. The surface bacteria adsorption was observed by scanning electron microscope (SEM). Result 1. SEM diagram showed that PU with high sulfonic ion content could reduce the adsorption of bacteria and sediment in bile circulation experiment, and there was no sediment formation in buffer cycling experiment, and there was no accumulation of bacteria adsorbed on the surface. Most of them are monolayer adsorbed and dispersed on the surface of material on average. In the analysis of bacterial adsorption capacity, it was shown that the adsorption capacity of surface bacteria decreased gradually with the increase of sulfonic acid group content. 2. After pretreatment with albumin, it was found that there was less bacterial adsorption on the surface of the material after albumin preadsorption in the buffer system, and with the increase of the preadsorption time of albumin, the amount of bacterial adsorption decreased. 3. After PE,OxPE,Pell-S-0.10 was preadsorbed by albumin, the inhibition effect of bacterial adsorption was good, and the amount of bacteria adsorbed by albumin preadsorbed for 60 minutes was less than that for 10 minutes, and the difference was statistically significant (P0.05), and in Pell-S-0.10, the amount of bacteria adsorbed by albumin preadsorbed in 60 minutes was lower than that in 10 minutes (P0.05). There was no significant difference in bacterial adsorption between 60 min and 10 min of albumin preadsorption. Conclusion the results suggest that the formation of bacterial biofilm and the adhesion of bile deposits on the surface of bile duct scaffolds can be reduced by polycarbamate sulfonate ionization and albumin preadsorption, which may be of some value in reducing the re-obstruction of biliary stents.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.7
，
本文编号：2273168
[Abstract]:Background obstructive jaundice is a kind of jaundice caused by gallstone or hepatobiliary tumor. Among many treatment methods, interventional artificial bile duct stent is an effective treatment method. Commercial polycarbamate artificial bile ducts are also used in clinical practice, and this material has good results in bile testing. However, the formation of biofilm in bile and the adhesion of sediments lead to re-obstruction of artificial bile duct stents. Therefore, changing the surface properties of bile duct scaffolds is an effective way to prevent re-clogging. Objective to study the effect of polycarbamate on bacterial adhesion, bile sediment and obstruction of bile duct scaffolds by surface modification of sulfonic groups and albumin adsorption. Method 1. Sodium (NaH) was added to PU solution to undergo bimolecular nucleophilic substitution reaction, followed by ring opening reaction with propane sulfate. Finally, PU, coating with different sulfonic ion content was produced in polyethylene (PE) and (OxPE) 2 oxide polyethylene tube. Select albumin concentration 1 mg / ml, preadsorption time 10 minutes and 60 minutes, the material of polyethylene, oxide polyethylene, sulfonated polyurethane, UV irradiation, connected in series, the two ends of the tube, Enter the buffer and stay in the tube for more than 12 hours. Aiming at different sulfonic ion content and protein adsorption for 10 minutes and 60 minutes of polycarbamate, the human bile and PBS containing 4-9 脳 107 CFU/mL DH5 a Escherichia coli were subjected to continuous cardiopulmonary bypass for 24 hours. 4. After 24 hours, the scaffold was washed with PBS, the first 3 cm of fluid inlet was discarded, then every 1 cm was sampled. The surface bacteria were immobilized in PBS solution containing 2.5% glutaraldehyde for 6 hours, and the drying process was carried out. The surface bacteria adsorption was observed by scanning electron microscope (SEM). Result 1. SEM diagram showed that PU with high sulfonic ion content could reduce the adsorption of bacteria and sediment in bile circulation experiment, and there was no sediment formation in buffer cycling experiment, and there was no accumulation of bacteria adsorbed on the surface. Most of them are monolayer adsorbed and dispersed on the surface of material on average. In the analysis of bacterial adsorption capacity, it was shown that the adsorption capacity of surface bacteria decreased gradually with the increase of sulfonic acid group content. 2. After pretreatment with albumin, it was found that there was less bacterial adsorption on the surface of the material after albumin preadsorption in the buffer system, and with the increase of the preadsorption time of albumin, the amount of bacterial adsorption decreased. 3. After PE,OxPE,Pell-S-0.10 was preadsorbed by albumin, the inhibition effect of bacterial adsorption was good, and the amount of bacteria adsorbed by albumin preadsorbed for 60 minutes was less than that for 10 minutes, and the difference was statistically significant (P0.05), and in Pell-S-0.10, the amount of bacteria adsorbed by albumin preadsorbed in 60 minutes was lower than that in 10 minutes (P0.05). There was no significant difference in bacterial adsorption between 60 min and 10 min of albumin preadsorption. Conclusion the results suggest that the formation of bacterial biofilm and the adhesion of bile deposits on the surface of bile duct scaffolds can be reduced by polycarbamate sulfonate ionization and albumin preadsorption, which may be of some value in reducing the re-obstruction of biliary stents.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.7
，
本文编号：2273168
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