Li Heng, Liu He, Zhang Lei, Hieawy Ahmed, Shen Ya
Department of Stomatology, Affiliated Hospital of Jining Medical University, Jining, China.
Division of Endodontics, Department of Oral Biological and Medical Sciences, Faculty of Dentistry, University of British Columbia, Vancouver, Canada.
J Dent Sci. 2023 Oct;18(4):1723-1730. doi: 10.1016/j.jds.2022.12.022. Epub 2023 Jan 11.
BACKGROUND/PURPOSE: Oral biofilms are highly structured bacterial colonies embedded in a highly hydrated extracellular polymeric substances (EPS) matrix. This study aimed to investigate the characteristics of oral biofilm at different stages of maturation.
Oral multispecies biofilms were grown anaerobically from plaque bacteria on collagen coated hydroxyapatite discs in brain heart infusion broth for one and three weeks. The volume of live bacteria and EPS matrix of the biofilms were determined by using corresponding fluorescent probes and confocal laser scanning microscopy. Atomic force microscopy (AFM) was used to quantitatively probe and correlate cell surface adhesion force of biofilms. The surface roughness was quantified in terms of the root mean square average of the height deviations. Adhesion was measured from force-distance data for the retraction of the cell from the surface.
The volume of live bacteria and EPS of 3-week-old biofilms was higher than 1-week-old biofilms. The surface roughness value in 1-week-old biofilms was significantly higher than that in 3-week-old biofilms. AFM force-distance curve results showed that the adhesion force at the cell-cell interface was significantly more at-tractive than those at bacterial cells surface of both stages biofilms. Adhesion forces between the AFM tip and the surface of bacterial cell were fairly constant, whereas the cell-cell interface experienced greater adhesion forces in the biofilm's development.
As oral biofilms become mature, EPS volume and cell-cell adhesion forces increase while the surface roughness decreases.
背景/目的:口腔生物膜是嵌入高度水合的细胞外聚合物(EPS)基质中的高度结构化细菌菌落。本研究旨在调查口腔生物膜在不同成熟阶段的特征。
在脑心浸液肉汤中,从牙菌斑细菌在胶原包被的羟基磷灰石圆盘上厌氧培养口腔多物种生物膜1周和3周。使用相应的荧光探针和共聚焦激光扫描显微镜测定生物膜中活细菌和EPS基质的体积。原子力显微镜(AFM)用于定量探测和关联生物膜的细胞表面粘附力。根据高度偏差的均方根平均值对表面粗糙度进行量化。从细胞从表面缩回的力-距离数据测量粘附力。
3周龄生物膜的活细菌和EPS体积高于1周龄生物膜。1周龄生物膜的表面粗糙度值显著高于3周龄生物膜。AFM力-距离曲线结果表明,两个阶段生物膜的细胞-细胞界面处的粘附力比细菌细胞表面处的粘附力明显更具吸引力。AFM针尖与细菌细胞表面之间的粘附力相当恒定,而在生物膜发育过程中,细胞-细胞界面经历了更大的粘附力。
随着口腔生物膜成熟,EPS体积和细胞-细胞粘附力增加,而表面粗糙度降低。