Oral Bio-Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong.
Arch Oral Biol. 2009 Nov;54(11):1052-60. doi: 10.1016/j.archoralbio.2009.08.002. Epub 2009 Aug 26.
The human fungal pathogen Candida is able to form biofilms in almost all the medical devices in current use. Indeed, biofilm formation is a major virulence attribute of microorganisms and account for a majority of human infections. Therefore, understanding processes appertaining to biofilm development is an important prerequisite for devising new strategies to prevent or eradicate biofilm-related infections. In the present study we used an array of both conventional and novel analytical tools to obtain a comprehensive view of Candida biofilm development. Enumeration of colony forming units, colorimetric (XTT) assay, Scanning Electron Microscopy (SEM) and novel Confocal Laser Scanning Microscopy (CLSM) coupled with COMSTAT software analyses were utilised to evaluate growth kinetics; architecture and viability of biofilms of a reference (ATCC) and a clinical strain each of two Candida species, C. albicans and C. glabrata. Biofilm growth kinetics on a polystyrene substrate was evaluated from the initial adhesion step (1.5 h) up to 72 h. These analyses revealed substantial inter- and intra-species differences in temporal organisation of Candida biofilm architecture, spatiality and cellular viability, while reaching maturity within a period of 48 h, on a polystyrene substrate. There were substantial differences in the growth kinetics upon methodology, although general trend seemed to be the same. Detailed architectural analysis provided by COMSTAT software corroborated the SEM and CSLM views. These analyses may provide a strong foundation for down stream molecular work of fungal biofilms.
人体真菌病原体念珠菌几乎能够在所有当前使用的医疗设备中形成生物膜。事实上，生物膜形成是微生物主要的毒力属性，也是大多数人类感染的原因。因此，了解生物膜形成过程是设计预防或消除生物膜相关感染的新策略的重要前提。在本研究中，我们使用了一系列传统和新颖的分析工具，以全面了解念珠菌生物膜的形成。通过使用平板计数、比色（XTT）测定、扫描电子显微镜（SEM）和新型共焦激光扫描显微镜（CLSM）结合 COMSTAT 软件分析，我们评估了参考株（ATCC）和两种念珠菌（白色念珠菌和光滑念珠菌）的临床株的生物膜生长动力学、生物膜结构和活力。在聚苯乙烯基质上评估了生物膜的生长动力学，从最初的粘附步骤（1.5 h）到 72 h。这些分析表明，念珠菌生物膜结构、空间和细胞活力的时间组织在种间和种内存在显著差异，而在聚苯乙烯基质上 48 h 内即可达到成熟。尽管总体趋势相同，但不同方法的生长动力学存在显著差异。COMSTAT 软件提供的详细结构分析与 SEM 和 CSLM 的观察结果相符。这些分析可以为真菌生物膜的下游分子工作提供坚实的基础。
