模拟人体膀胱体外构建仿生尿流应力细菌生物膜动态孵育系统  

Simulating the human bladder to construct a bacterial biofilm dynamic incubation system under biomimetic urinary flow stress in vitro

作　　者：安宇[1] 邱明星[1] 刘竞[1] 黄润华 王寓 王世泽 An Yu;Qiu Mingxing;Liu Jing;Huang Runhua;Wang Yu;Wang Shize(Department of Urology,Sichuan Academy of Medical Sciences&Sichuan Provincial People’sHospital)

机构地区：[1]四川省医学科学院·四川省人民医院泌尿外科,成都610072

出　　处：《重庆医科大学学报》2023年第10期1195-1200,共6页Journal of Chongqing Medical University

基　　金：四川省自然科学基金资助项目(编号:23NSFSC1558);四川省卫生健康委员会科研资助项目(编号:18PJ454);四川省医学科学院临床研究及转化基金资助项目(编号:2018LY11)。

摘　　要：目的:模拟人体膀胱体外构建仿生尿流应力细菌生物膜动态孵育系统,探讨该系统的应用效果。方法:体外构建细菌生物膜动态孵育系统,采用大肠埃希菌为实验菌,硅胶膜片为成膜载体,建立5种尿流应力输出:生理白天和夜间分泌、生理白天排尿、病理白天和夜间排尿;设置2组仿生尿流应力流转方案:生理组和病理组。观察仿生尿流应力刺激下大肠埃希菌生物膜形成特点。检测方法包括:生物膜细菌活菌计数,荧光染色结合激光共聚焦显微镜观察,测定生物膜细菌荧光图像平均光密度值,构建生物膜立体图测量其厚度。测值多组间均数比较采用单因素方差分析(one-way analysis of variance,ANOVA),组内均数两两比较采用LSD-t法。结果:各组内比较结果显示,随孵育时间延长,生物膜细菌活菌计数值均逐渐增加,病理组48 h、静态组和生理组60 h显著增加并达到峰值(cP=0.000);生物膜细菌荧光图像平均光密度值和生物膜厚度测值均逐渐增加,病理组48 h、静态组和生理组72 h显著增加并达到峰值(cP=0.000);各组内其余测值差异无统计学意义(P>0.01)。各组间比较结果显示,与静态组比较,生理组和病理组36 h、48 h、60 h、84 h、96 h生物膜细菌活菌计数值均显著增加(aP=0.000),与生理组比较,病理组增加更为显著(bP=0.000);与静态组比较,生理组96 h,病理组48 h、96 h生物膜细菌荧光图像平均光密度值显著增多(aP=0.000),生理组72 h,病理组48 h、96 h BF厚度测值显著增多(aP=0.000);与生理组比较,病理组48 h、96 h细菌荧光图像平均光密度值和生物膜厚度测值均显著增多(bP=0.000);各组间其余测值差异无统计学意义(P>0.01)。结论:本研究模拟人体膀胱体外成功构建细菌生物膜动态孵育系统,经验证仿生尿流应力显著促进大肠埃希菌生物膜形成。Objective:To construct a dynamic incubation system of biomimetic urinary flow stress bacteria biofilm by simulating hu-man bladder in vitro,and to explore the application effect of this system.Methods:For the dynamic incubation system to construct bac-terial biofilm in vitro,we used Escherichia coli as the experimental bacteria and silica gel membrane as the carrier of membrane forma-tion to establish five bionic effects of urinary flow stress:physiological secretion during the day and night,physiological urination dur-ing the day,and pathological urination during the day and night.In addition,we set up two circulation scheme groups:physiological group and pathological group to observe the characteristics of biofilm formation of Escherichia coli under the stimulation of bionic uri-nary flow stress.The detection methods included:counting the living bacteria of biofilm bacteria,fluorescence staining combined with laser confocal microscope observation,determining the average light density value of the fluorescence image of biofilm bacteria,and con-structing biofilm stereogram to measure the thickness of biofilm.The average values among multiple groups were detected by using one-way analysis of variance(ANOVA),and LSD-t method was used to compare the average values within groups in pairs.Results:Through the comparison of data in each group,with the prolongation of incubation time,the number of viable bacteria of biofilm bacteria increased gradually,and the number of viable bacteria in pathologi-cal group at 48 h,static group and physiological group at 60 h increased significantly and reached the peak(cP=0.000);The measured values of average light density and biofilm thickness in the fluorescence image of biofilm bacteria increased gradually,and the mea-sured values in the pathological group at 48 h,the static group and the physiological group at 72 h increased significantly and reached the peak(cP=0.000);There was no significant difference in other measured values in each group(P>0.01).Through the com-parison of data

关 键 词：膀胱 大肠埃希菌 细菌生物膜 生物学 尿流应力 动态孵育 

分 类 号：R378[医药卫生—病原生物学]