Ohsumi Tatsuya, Takenaka Shoji, Wakamatsu Rika, Sakaue Yuuki, Narisawa Naoki, Senpuku Hidenobu, Ohshima Hayato, Terao Yutaka, Okiji Takashi
Division of Cariology, Operative Dentistry and Endodontics, Department of Oral Health Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Department of Food Bioscience and Biotechnology, College of Bioresource Sciences, Nihon University, Kanagawa, Japan.
PLoS One. 2015 Jan 30;10(1):e0116647. doi: 10.1371/journal.pone.0116647. eCollection 2015.
Chemical disinfection of oral biofilms often leaves biofilm structures intact. This study aimed to examine whether the residual structure promotes secondary bacterial adhesion. Streptococcus mutans biofilms generated on resin-composite disks in a rotating disc reactor were disinfected completely with 70% isopropyl alcohol, and were again cultured in the same reactor after resupplying with the same bacterial solution. Specimens were subjected to fluorescence confocal laser scanning microscopy, viable cell counts and PCR-Invader assay in order to observe and quantify secondarily adhered cells. Fluorescence microscopic analysis, particularly after longitudinal cryosectioning, demonstrated stratified patterns of viable cells on the disinfected biofilm structure. Viable cell counts of test specimens were significantly higher than those of controls, and increased according to the amount of residual structure and culture period. Linear regression analysis exhibited a high correlation between viable and total cell counts. It was concluded that disinfected biofilm structures favored secondary bacterial adhesion.
口腔生物膜的化学消毒通常会使生物膜结构保持完整。本研究旨在检验残留结构是否会促进细菌二次黏附。在旋转圆盘反应器中树脂复合材料圆盘上形成的变形链球菌生物膜用70%异丙醇完全消毒,在重新加入相同细菌溶液后,再次在同一反应器中培养。对样本进行荧光共聚焦激光扫描显微镜检查、活菌计数和PCR-入侵者检测,以观察和定量二次黏附的细胞。荧光显微镜分析,尤其是纵向冷冻切片后的分析,显示了消毒生物膜结构上活细胞的分层模式。测试样本的活菌计数显著高于对照组,并根据残留结构的量和培养时间而增加。线性回归分析显示活菌计数与总细胞计数之间存在高度相关性。得出的结论是,消毒后的生物膜结构有利于细菌二次黏附。