Spittaels Karl-Jan, Coenye Tom
Laboratory of Pharmaceutical Microbiology, Ghent University, Ghent, Belgium.
Laboratory of Pharmaceutical Microbiology, Ghent University, Ghent, Belgium.
Anaerobe. 2018 Feb;49:21-29. doi: 10.1016/j.anaerobe.2017.11.002. Epub 2017 Nov 22.
The aim of the present study was to develop a new model system to study Propionibacterium acnes biofilms. This model should be representative for the conditions encountered in the pilosebaceous unit.
The new model, consists of an artificial sebum pellet supported by a silicone disc. Sebum pellets were inoculated with various P. acnes strains isolated from both normal and acneic skin. Growth and biofilm formation was verified by conventional plating at different time points, as well as by resazurin assays and fluorescence microscopy after LIVE/DEAD staining. The artificial sebum pellets were also used in assays to measure the production of certain virulence factors implicated in the pathogenesis of acne, including lipase, protease and the presence of CAMP factors.
The artificial sebum model can sustain biofilm growth of P. acnes, as was determined by increasing CFU counts for up to 1 week after inoculation. Metabolic activity and biofilm formation were confirmed using resazurin staining and fluorescence microscopy respectively. The production of virulence factors in this model was demonstrated as well.
本研究的目的是开发一种新的模型系统来研究痤疮丙酸杆菌生物膜。该模型应能代表毛囊皮脂腺单位中遇到的情况。
新模型由硅树脂圆盘支撑的人工皮脂颗粒组成。用从正常皮肤和痤疮皮肤分离出的各种痤疮丙酸杆菌菌株接种皮脂颗粒。通过在不同时间点进行常规平板培养,以及在活/死染色后通过刃天青测定法和荧光显微镜检查来验证生长和生物膜形成。人工皮脂颗粒还用于测定某些与痤疮发病机制有关的毒力因子的产生,包括脂肪酶、蛋白酶和CAMP因子的存在。
人工皮脂模型能够维持痤疮丙酸杆菌的生物膜生长,接种后长达1周的CFU计数增加证明了这一点。分别使用刃天青染色和荧光显微镜检查证实了代谢活性和生物膜形成。该模型中也证明了毒力因子的产生。