Univ of Campinas-UNICAMP, Piracicaba Dental School, Department of Prosthodontics and Periodontology, Piracicaba, SP, Brazil.
Braz Oral Res. 2013 Jul-Aug;27(4):369-75. doi: 10.1590/S1806-83242013005000020.
The purpose of this study was to evaluate the influence of the substratum position and the saliva acquired pellicle (AP) on Candida albicans biofilm development. Poly(methylmethacrylate) (PMMA) disks were fabricated and randomly allocated to experimental groups: HNP (disks placed in a horizontal position and uncoated by pellicle), VNP (disks placed in a vertical position and uncoated by pellicle), HCP (disks placed in a horizontal position and coated by pellicle), and VCP (disks placed in a vertical position and coated by pellicle). Disks were placed in a 24-well plate and a suspension of 107 cells/mL of Candida albicans was added to each well for biofilm development. The plates were aerobically incubated at 35°C. The biofilms were evaluated at 1.5 (adhesion time point), 24, 48, 72, and 96 hours. The number of viable cells was quantified in terms of the colony-forming units per milliliter (CFU/mL). Metabolic activity was measured by the XTT assay. The biofilm structure was analyzed by scanning electron microscopy. The data were analyzed by three-way ANOVA followed by Tukey's test, with significance set at 5%. The vertical groups showed less biofilm formation and lower metabolic activity than the horizontal groups (p<0.05). Significant differences in cell viability and metabolic activity were observed between the adhesion and other time points (p<0.05), but these variables were not affected by the presence of the pellicle (p>0.05). It can be concluded that the substratum position influenced biofilm development.
本研究旨在评估基底位置和唾液获得的黏膜(AP)对白色念珠菌生物膜形成的影响。制备聚甲基丙烯酸甲酯(PMMA)圆盘,并将其随机分配到实验组:HNP(放置在水平位置且未被黏膜覆盖的圆盘)、VNP(放置在垂直位置且未被黏膜覆盖的圆盘)、HCP(放置在水平位置且被黏膜覆盖的圆盘)和 VCP(放置在垂直位置且被黏膜覆盖的圆盘)。圆盘放置在 24 孔板中,向每个孔中加入 107 个细胞/mL 的白色念珠菌悬浮液以进行生物膜形成。平板在 35°C 下有氧孵育。在 1.5 小时(黏附时间点)、24、48、72 和 96 小时评估生物膜。以每毫升菌落形成单位(CFU/mL)的形式定量活细胞数。通过 XTT 测定法测量代谢活性。通过扫描电子显微镜分析生物膜结构。数据通过三因素方差分析(ANOVA)和 Tukey 检验进行分析,置信水平设为 5%。垂直组的生物膜形成和代谢活性低于水平组(p<0.05)。在黏附时间点和其他时间点之间观察到细胞活力和代谢活性的显著差异(p<0.05),但这些变量不受黏膜的存在影响(p>0.05)。可以得出结论,基底位置影响生物膜的形成。