Department of Biology and South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, Texas 78249, USA.
J Clin Microbiol. 2009 Dec;47(12):4078-83. doi: 10.1128/JCM.01377-09. Epub 2009 Sep 30.
Urinary tract infections (UTIs) are the most common type of nosocomial infection, and Candida albicans is the most frequent organism causing fungal UTIs. Presence of an indwelling urinary catheter represents a significant risk factor for UTIs. Furthermore, these infections are frequently associated with the formation of biofilms on the surface of these catheters. Here, we describe the characterization of C. albicans biofilms formed in vitro using synthetic urine (SU) medium and the frequently used RPMI medium and compare the results. Biofilms of C. albicans strain SC5314 were formed in 96-well microtiter plates and on silicon elastomer pieces using both SU and RPMI media. Biofilm formation was monitored by microscopy and a colorimetric XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction assay. As in biofilms grown in RPMI medium, time course studies revealed that biofilm formation using SU medium occurred after an initial adherence phase, followed by growth, proliferation, and maturation. However, microscopy techniques revealed that the architectural complexity of biofilms formed in SU medium was lower than that observed for those formed using RPMI medium. In particular, the level of filamentation of cells within the biofilms formed in SU medium was diminished compared to those in the biofilms grown in RPMI medium. This observation was also corroborated by expression profiling of five filamentation-associated genes using quantitative real-time reverse transcriptase PCR. Sessile C. albicans cells were resistant to fluconazole and amphotericin B, irrespective of the medium used to form the biofilms. However, caspofungin exhibited potent in vitro activity at therapeutic levels against C. albicans biofilms grown in both SU and RPMI media.
尿路感染(UTIs)是最常见的医院获得性感染类型,而白色念珠菌是引起真菌性 UTIs 的最常见病原体。留置导尿管的存在是 UTIs 的一个重要危险因素。此外,这些感染通常与这些导管表面生物膜的形成有关。在这里,我们描述了使用合成尿液(SU)培养基和常用的 RPMI 培养基在体外形成的白色念珠菌生物膜的特征,并比较了结果。使用 SU 和 RPMI 培养基,在 96 孔微量滴定板和硅弹性体片上形成了白色念珠菌菌株 SC5314 的生物膜。通过显微镜和比色 XTT [2,3-双(2-甲氧基-4-硝基-5-磺苯基)-2H-四唑-5-羧基苯胺]还原测定监测生物膜的形成。与在 RPMI 培养基中生长的生物膜一样,时间过程研究表明,使用 SU 培养基形成生物膜后经过初始附着阶段,然后是生长、增殖和成熟。然而,显微镜技术显示,SU 培养基中形成的生物膜的结构复杂性低于 RPMI 培养基中观察到的生物膜。特别是,与在 RPMI 培养基中生长的生物膜相比,SU 培养基中形成的生物膜内细胞的丝状化程度降低。使用定量实时逆转录 PCR 对五个丝状相关基因的表达谱进行分析也证实了这一观察结果。无论使用哪种培养基形成生物膜,静止的白色念珠菌细胞对氟康唑和两性霉素 B 均具有抗性。然而,卡泊芬净在治疗水平下对 SU 和 RPMI 培养基中生长的白色念珠菌生物膜均表现出强大的体外活性。