牙龈卟啉单胞菌临床菌株荚膜相关的表面性质分析  

Analysis of capsule related surface properties of Porphyromonas gingivalis isolates

作　　者：周彦玢[1] 刘大力[1] 李云鹏[1] 王依玮[1] 束蓉[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔医学院牙周病科上海市口腔医学研究所上海市口腔医学重点实验室,200011

出　　处：《中华口腔医学杂志》2014年第3期145-150,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（30901672）;高等学校博士学科点专项科研基金（20090073120096）;上海市科学技术委员会科研计划（12JC1407502）

摘　　要：目的 分析牙龈卟啉单胞菌（Porphyromonas gingivalis,Pg）临床菌株的多种表面性质,初步探讨Pg菌株荚膜与其表面性质的关系.方法 利用透射电镜观测5株临床菌株（SJD2、SJD12、SJD4、SJD5和SJD11）和2株模式菌株（W83、ATCC33277）的荚膜结构和厚度,采用微生物烃类黏附实 验检测受试菌株的表面亲疏水性质,并使用自凝集实验观察受试菌株浮游生长的能力.采用生物膜半定量检测和激光扫描共聚焦显微镜检测各菌株形成生物膜的能力.应用Student-Newman Kewls检验对结果数据进行组间两两比较,以双侧P＜0.05为差异有统计学意义.结果 Pg菌株的荚膜厚度各异,高毒模式菌株W83的荚膜厚度[（25.11 ±1.18） nm]显著大于低毒模式菌株ATCC33277[（14.87±1.01） nm] （P ＜0.05）.临床菌株的荚膜厚度由厚至薄依次为：SJD4、SJD11、SJD5、SJD2、SJD12.荚膜较厚的W83、SJD4、SJD11菌株疏水率低,细菌不易自凝集,呈浮游生长;荚膜较薄的ATCC33277、SJD5、SJD2、SJD12菌株疏水率较W83显著上升,且差异有统计学意义（P＜0.05）,自凝集现象明显.生物膜半定量检测实验中,除SJD2和SJD4的生物膜染色后具有相同水平的吸光度之外,其余菌株吸光度差异均有统计学意义（P＜0.05）,各菌株生物膜中的细菌量并不与荚膜厚度的差异相吻合.各菌株生物膜厚度从（14.74 ±4.99）至（24.13 ±5.45） μm不等,差异均无统计学意义（p＞0.05）.激光扫描共聚焦显微镜观察可见W83、SJD11生物膜内荧光信号较弱,生物膜密度较低,其他菌株可生成较致密且荧光信号较强的生物膜结构,存在空隙或孔道结构.结论 Pg菌株表面性质的多样性与荚膜厚度存在一定关联.Objective To analyze the capsule related surface properties of Porphyromonas gingivalis （Pg） isolates.Methods The transmission electron microscopy （TEM） was used to observe the capsule structure and the capsule thickness of 2 type of strains and 5 clinical isolates.Microbial adhesion to hydrocarbons（MATH） assay was used to qualitatively assess the hydrophobicity of each strain,and the capacities of these strains were investigated by autoaggregation assay.Ninety-six well biofilm assay and confocal laser scanning microscopy （CLSM） were applied to quantify and observe the biofilm produced by each strain.Results TEM showed the variety of capsule thickness of these strains.Virulent type strain W83 possessed thicker capsular structure than less-virulent type strain ATCC33277.The SJD4 possessed thicker capsule than other clinical isolates,followed by SJD11,SJD5,SJD2,and SJD12.Strains W83,SJD4,SJD11,with thicker capsule,were much more hydrophilic with lower MATH percentage,in accordance with a slow autoaggregation in incubation during a period of 240 min.Compared with W83,the hydrophobicity of strains ATCC33277,SJD5,SJD2,and SJD12,with thinner capsule,showed increased MATH percentage and autoaggregations.All clinical strains developed biofilm with different absorbance compared with type strains.The CLSM observation showed biofilm thickness of each strain,ranged from （14.74 ± 4.99） to （24.13-± 5.45） μm.Strain W83 and SJD11 showed notable poor biofilm formation,while others developed dense and mature biofilm.Conclusions There was a certain degree of linkage between the Pg capsule thickness and surface properties diversity.

关 键 词：紫单胞菌 龈 细菌荚膜 临床菌株 表面性质 

分 类 号：R781.4[医药卫生—口腔医学]