【摘要】 目的研究高强度聚焦超声(HIFU)对体外铜绿假单胞菌(PA)生物被膜(BF)杀菌作用和对其结构的影响。方法体外建立铜绿假单胞菌PAO1生物被膜3、6 d模型,平板计数法菌落进行计数,FITC-ConA染胞外多糖(EPS),SYTO9/PI标记BF内细菌、激光共聚焦显微镜摄取不同层面的图像,采用Image Pro Plus 6.0(IPP)分析死菌比例和运用BF图像结构分析软件(ISA)进行定量分析。结果 (1)3 d和6 d组菌落数经HIFU干预后减少。(2)荧光显微镜下观察可见HIFU干预后EPS减少。(3)3 d和6 d组死菌比例经HIFU干预后增加。(4)ISA结果3 d和6 d组均见BF厚度减少,区域孔率增加,平均扩散距离减小,结构熵减小。结论经HIFU干预后PA BF中死菌比例增加和BF空间结构破坏。更多还原

【Abstract】 Objective To investigate the effects of high intensity focused ultrasound(HIFU) on bactericidal action and the structure of Pseudomonas aeruginosa biofilm in vitro.Methods Models of 3 d,6 d Pseudomonas aeruginosa PAO1 biofilm were established in vitro.Determining the number of colony forming units(CFU) was adopted. Exopolysaccharides(EPS) of the biofilm was stained by FITC-ConA.Confocal Laser Scanning Microscope(CLSM) was used,based on fluorophores from by Image Pro Plus 6.0.Quantification of the effect could be calculated by Image Structural Analyzer(ISA) software.Results(1) Colony forming units(CFU) of 3 d and 6 d models were reduced after intervention.(2)Exopolysaccharides(EPS) were reduced in some way.(3)Bactercial mortality rate of 3 d and 6 d models were increased after intervention.(4)Parameters of image structural analyzer(ISA);after intervention biofilm thickness of 3 d and 6 d models were decreased;the areal porosity(AP) were increased,the average diffusion distance (ADD) were reduced,the textual entropy(TE) were reduced.Conclusions The bacterial mortality rate increases in some way and the structure of biofilm could be destroyed by HIFU.更多还原