Effect of Nystatin on Candida albicans- Streptococcus mutans duo-species biofilms
Effect of Nystatin on Candida albicans- Streptococcus mutans duo-species biofilms
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DOI：
10.1016/j.archoralbio.2022.105582
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发表时间：
2022-11-14
影响因子：
3
通讯作者：
Xiao, Jin
中科院分区：
文献类型：
--
作者：
Alomeir, Nora;Zeng, Yan;Xiao, Jin
目的：采用体外致龋性生物被膜模型评价制霉菌素对白色念珠菌和变形链球菌双菌种生物膜的影响。设计：在高糖(1%蔗糖和1%葡萄糖)条件下，在唾液包被的羟基磷灰石片上形成生物膜，接种105CFU/ml变形链球菌和103CFU/ml白色念珠菌。在20~68h期间，用28000IU制霉菌素溶液处理生物膜，5min/次，4次/d，模拟临床应用。利用多光子共聚焦显微镜对生物膜的三维结构进行了评价。用实时荧光定量聚合酶链式反应检测白念珠菌和变形链球菌毒力基因的表达。在3个独立的重复中使用重复的光盘。结果：制霉菌素治疗44h后，生物膜中白色念珠菌均被清除，治疗后44h和68h，制霉菌素组生物被膜干重明显减少，变形链球菌数量减少0.5logcfu/ml(p<；0.05)。值得注意的是，制霉菌素处理改变了垂直布局上的生物量分布，导致与对照相比，制霉菌素处理生物膜基质层上的体积更小。在制霉菌素处理的生物膜中也观察到微集落大小和体积的减少(p<；0.05)。制霉菌素处理的生物膜形成了独特的晕状微菌落，减少了核心EPS的覆盖率。此外，制霉菌素生物膜显著下调了变形链球菌gtfd和atpD基因的表达(p<；0.05)。结论：制霉菌素的应用改变了白色念珠菌和变形链球菌双菌种生物膜的形成和特征。因此，从抗真菌的角度开发预防或治疗龋齿的临床方案是必要的。
Objective: To assess the effect of Nystatin on Candida albicans and Streptococcus mutans duo-species biofilms using an in vitro cariogenic biofilm model.Design: Biofilms were formed on saliva-coated hydroxyapatite discs under high sugar challenge (1 % sucrose and 1 % glucose), with inoculation of 105CFU/ml S. mutans and 103CFU/ml C. albicans. Between 20 and 68 h, biofilms were treated with 28,000 IU Nystatin solution, 5 min/application, 4 times/day, to mimic the clinical application. Biofilm's three-dimensional structure was assessed using multi-photon confocal microscopy. The expression of C. albicans and S. mutans virulence genes was assessed via real-time PCR. Duplicate discs were used in 3 independent repeats. t-test and Mann-Whitney U test were used to compare outcomes between treatment and control group.Results: Nystatin treatment eliminated C. albicans in biofilms at 44 h. Nystatin-treated group had a significant reduction of biofilm dry-weight and reduced S. mutans abundance by 0.5 log CFU/ml at 44 and 68 h (p < 0.05). Worth noting that biomass distribution across the vertical layout was altered by Nystatin treatment, resulting in less volume on the substrate layers in Nystatin-treated biofilms compared to the control. Reduction of micro-colonies size and volume was also observed in Nystatin-treated biofilms (p < 0.05). Nystatin-treated biofilms formed unique halo-shaped microcolonies with reduced core EPS coverage. Furthermore, Nystatin-treated bio-films had significant down-regulations of S. mutans gtfD and atpD genes (p < 0.05).Conclusions: Nystatin application altered the formation and characteristics of C. albicans and S. mutans duo -species biofilms. Therefore, developing clinical regimens for preventing or treating dental caries from an anti-fungal perspective is warranted.