隐球菌生物膜的构建、结构及影响因素的研究

目的：构建隐球菌生物膜体外、动物模型，观察其形成的特点、超微结构，脑脊液及血清对生物膜活性的影响。方法：采用小鼠腹腔内置凹槽法，构建隐球菌生物膜动物模型。采用96孔微量培养皿构建不同血清型隐球菌生物膜体外模型，Vit k3增强的四甲基偶氮唑盐（MTT）法，评价隐球菌生物膜的体外生长动力学，脑脊液及血清对隐球菌生物膜活性的影响；光镜、电镜和共焦激光扫描显微镜观察隐球菌生物膜超微结构。结果：构建隐球菌生物膜体外模型和动物模型；隐球菌生物膜活性随培养时间延长而增强，脑脊液和血清具有明显增强隐球菌生物膜活性的作用，不同血清型生成生物膜的厚度和其生长动力学不同。结论：隐球菌可以产生生物膜；脑脊液和血清具有明显增强其生物膜活性的作用，不同血清型生成隐球菌生物膜的能力不同。

Cryptococcus biofilm: constrction, formation, and influencing factors

Objective:To construct Cryptococcus biofilm model in vitro and in vivo and to observe its formation,ultrastructure and the influences of serum and CSF(cerebrospinal fluid)on the biofilm.Methods:Cryptococcus biofilm model in vivo by laying groove in mice enterocoelia and the Cryptococcus biofilm model in vitro in 96-well microculture plates were constructed,three-dimensional structure of Cryptococcus biofilm being observed with scanning laser confocal microscopy(LSCM).MTT was used to evaluate the growth kinetics of biofilm at various time points during biofilm formation and its influences by serum and CSF.Results:Mature Cryptococcus biofilms,followed by growth,proliferation,and maturation,consisted of a dense network of yeasts cells displayed spatial heterogeneity and the structure of microcolonies with ramifying water channels.The metabolic activity of Cryptococcus biofilm increased during biofilm formation,and serum and CSF could obviously increase the activity of biofilm.Conclusion:Cryptococcus could form biofilm.Serum and CSF could increase the activity of the biofilm.