【摘要】：背景梗阻性黃疸是由膽結(jié)石或肝膽部位的腫瘤所造成的黃疸,在眾多的治療方法中,介入人工膽管支架是一種有效的治療方法；商業(yè)化聚氨基甲酸酯材質(zhì)的人工膽管也在臨床上應(yīng)用,且此材質(zhì)在膽汁的測試中有不錯的成效,然而膽汁中生物膜的形成以及沉積物的附著導(dǎo)致人工膽管支架再阻塞,從而縮短了使用期限,因此,改變膽管支架材質(zhì)的表面性質(zhì)是防止再堵塞的有效方法。目的通過對聚氨基甲酸酯作磺酸根離子基團(tuán)的改質(zhì),同時以白蛋白吸附進(jìn)行表面處理,探討對膽管支架表面細(xì)菌粘附、膽汁沉積物和膽管支架堵塞的影響。方法1. 將氫化鈉(NaH)加入PU溶液進(jìn)行雙分子親核取代反應(yīng),之后再加上丙烷硫酸酯進(jìn)行開環(huán)反應(yīng),最后產(chǎn)生不同磺酸根離子含量的PU,涂層于聚乙烯(PE)及氧化聚乙烯管(OxPE)2.選擇白蛋白濃度1mg/ml,預(yù)吸附時間10分鐘與60分鐘,將材質(zhì)為聚乙烯、氧化聚乙烯、磺酸化聚氨酯、基聚氨酯的支架經(jīng)紫外線照射,以串聯(lián)方式連接,兩端接三通管,通入緩沖液并在管內(nèi)停留12小時以上3. 針對不同磺酸根離子含量及蛋白質(zhì)吸附10分鐘及60分鐘的聚氨基甲酸酯,在濃度為4-9×107 CFU/mL DH5 a型大腸桿菌的人體膽汁及PBS中做體外24小時連續(xù)循環(huán)實(shí)驗。4. 實(shí)驗經(jīng)24小時后,將支架以PBS潤洗,流體入口的前3厘米舍棄,之后每1厘米取樣一次,共取三次進(jìn)行表面吸附細(xì)菌的定量,另外再取1厘米,使用含2.5%戊二醛的PBS溶液進(jìn)行表面細(xì)菌的固定化6小時,并進(jìn)行干燥處理程序。接著材質(zhì)進(jìn)行鍍金處理,以掃描式電子顯微鏡(SEM)進(jìn)行表面細(xì)菌吸附情形的觀察。結(jié)果1. SEM圖可觀察到在膽汁循環(huán)實(shí)驗方面,高磺酸根離子含量的PU能減少細(xì)菌及沉積物的吸附；在緩沖液循環(huán)實(shí)驗方面,沒有沉積物的生成,且表面吸附的細(xì)菌沒有聚集的現(xiàn)象,大多呈單層吸附,并平均分散在材質(zhì)的表面。在細(xì)菌吸附量分析方面,顯示隨著磺酸基含量的增加,表面細(xì)菌的吸附量逐漸減少。2. 材質(zhì)經(jīng)白蛋白的預(yù)吸附處理后,在緩沖液系統(tǒng)下,發(fā)現(xiàn)經(jīng)白蛋白預(yù)吸附后的材質(zhì)表面有較少的細(xì)菌吸附,且隨白蛋白預(yù)吸附時間增加,細(xì)菌吸附量有減少的趨勢。3. PE、OxPE、Pell-S-0.10經(jīng)白蛋白預(yù)吸附處理后,抑制細(xì)菌吸附的效果好；且在P E、OxPE中,白蛋白預(yù)吸附60分鐘比10分鐘其細(xì)菌吸附量更少,其差異具有統(tǒng)計學(xué)意義(P0.05),在Pell-S-0.10中,白蛋白預(yù)吸附60分鐘與10分鐘細(xì)菌吸附量無明顯差異。結(jié)論本實(shí)驗顯示聚氨基甲酸酯磺酸根離子化及白蛋白預(yù)吸附能減少膽管支架表面細(xì)菌生物膜的形成以及膽汁沉積物的附著,對減少膽管支架再阻塞可能有一定價值。
[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.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.7
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本文編號：2273168