雌二醇对乳腺癌假体植入术后表皮葡萄球菌生物膜形成的影响研究  

EFFECT OF ESTRADIOL ON BIOFILM FORMATION OF STAPHYLOCOCCUS EPIDERMIDIS AFTER BREAST IMPLANT SURGERY

作　　者：王曦[1] 黄云超[2] 周永春 刘德权[1] 吕志勇[4] 叶联华[2] 陈颖[2] 杨加鹏[2] 成鹏[2] 王正玺 

机构地区：[1]昆明医科大学第三附属医院/云南省肿瘤医院乳腺外一科,昆明650118 [2]昆明医科大学第三附属医院/云南省肿瘤医院胸外一科,昆明650118 [3]云南省肺癌研究重点实验室 [4]昆明医科大学第三附属医院/云南省肿瘤医院麻醉科,昆明650118

出　　处：《中国修复重建外科杂志》2016年第7期876-884,共9页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金面上项目(30872555);国家自然科学基金重点项目(U1202224);云南省卫生厅科技计划项目(2012WS041)

摘　　要：目的探讨雌二醇对乳腺癌术后假体表面表皮葡萄球菌生长以及生物膜形成能力及其结构的影响。方法取表皮葡萄球菌标准株ATCC35984,调整浓度至1×107 CFU/m L或1×108 CFU/m L,与硅胶片和终浓度为125 pmol/L的雌二醇混合培养,制备硅胶材料表面细菌生物膜体外模型。于培养后4、6、12、24、48、72 h,采用结晶紫染色半定量检测硅胶材料表面细菌生物膜形成能力,XTT法检测硅胶材料表面细菌生长动力;根据以上检测结果,选择细菌悬液实验浓度。取实验浓度表皮葡萄球菌ATCC35984悬液以及终浓度为50、125、250、500 pmol/L的雌二醇悬液,分别与硅胶片混合培养制备生物膜体外模型,以未添加雌二醇悬液(0 pmol/L)作为对照;另取实验浓度表皮葡萄球菌ATCC12228悬液,同法制备模型,作为阴性对照。于培养后4、6、12、24、48、72 h同上法检测硅胶材料表面细菌生长动力及生物膜形成能力,扫描电镜观察硅胶材料表面细菌生物膜超微结构;培养6、12、24 h激光共聚焦显微镜观察硅胶材料表面细菌生物膜厚度。结果根据XTT法及结晶紫染色半定量检测结果,选择浓度为1×107 CFU/m L细菌悬液进行实验。XTT法检测示,ATCC12228和ATCC35984菌株分别在4、6、12、24、72 h和4、6、24、72 h时,其125、250、500 pmol/L组细菌生长速度快于0、50 pmol/L组(P<0.05),4、6、72 h时500 pmol/L组快于125、250 pmol/L组(P<0.05),72 h时250 pmol/L组快于125 pmol/L组(P<0.05),其余组间比较差异无统计学意义(P>0.05);同一时间点两种菌株相同雌二醇浓度组比较,差异均无统计学意义(P>0.05)。结晶紫染色半定量检测:各时间点ATCC12228菌株各雌二醇浓度组生物膜形成均为阴性。而ATCC35984菌株培养4 h即有生物膜形成,随时间延长逐渐增厚,24 h时达峰值;培养4、6 h,0 pmol/L组生物膜厚度大于125、250、500 pmol/L组(P<0.05);12 h开始125 pmol/L组生物膜最厚,与其他组比较差异均有统�Objective To investigate the effect of the estradiol hormones on biofilm formati on and structure of Staphylococcus epidermidis after breast implant surgery. Methods The concentration of Staphylococcus epidermidis strains ATCC35984 was adjusted to 1×10^7 CFU/m L or 1×10^8 CFU/m L, and the type strains were incubated on the surface of silica gel in 125 pmol/L estradiol suspensions to prepare bacterial biofilms model in vitro. After cultured in vitro for 4, 6, 12, 24, 48, and 72 hours, bacteria growth and biofilm formation ability were assessed by means of the XTT and crystal violet staining respectively. According to the above results, the bacterial suspension concentration was selected for experiments. The experimental concentration of Staphylococcus epidermidis ATCC35984 suspension and the concentrations of 50, 125, 250, 500 pmol/L estradiol suspensions were mixed with silica gel respectively to prepare biofilm model in vitro, no estradiol suspension served as control group. The experimental concentration of Staphylococcus epidermidis ATCC12228 suspension was used to prepare the same model in the negative control. After cultured in vitro for 4, 6, 12, 24, 48, and 72 hours, the same methods were used to assess the bacteria growth dynamics and biofilm forming ability, and the scanning electron microscope（SEM） was used to observe bacterial biofilm structure cultured on the surface of silica gel; the laser scanning confocal microscope（CLSM） was used to measure bacterial biofilm thickness on the surface of silica gel after 6, 12, and 24 hours. Results According to the results of semi quantitative detection of crystal violet stain and XTT methods, the bacterial suspension of 1×10^7 CFU/m L was selected for the experiment. XTT results indicated that the growth rates of ATCC12228 strain（at 4, 6, 12, 24, and 72 hours） and ATCC35984 strain（at 4, 6, 24, and 72 hours） in 125, 250, and 500 pmol/L estradiol were significantly faster than those in 0 and 50 pmol/L（P〈0.05）. The growth rate of 500 pmol/L

关 键 词：雌二醇 乳房假体 乳腺癌 表皮葡萄球菌 细菌生长 细菌生物膜 

分 类 号：R737.9[医药卫生—肿瘤]