Department of Dental Pathology, Operative Dentistry and Endodontics, School of Dentistry, Aarhus Faculty of Health Sciences, Aarhus University, Denmark.
Acta Odontol Scand. 2012 Dec;70(6):441-7. doi: 10.3109/00016357.2011.634833. Epub 2011 Nov 30.
The study of initial microbial colonization on dental surfaces is a field of intensive research because of the aetiological role of biofilms in oral diseases. Most previous studies of de novo accumulation and composition of dental biofilms in vivo do not differentiate between biofilms formed during day and night. This study hypothesized that there is a diurnal variation in the rate of accumulation of bacteria on solid surfaces in the oral cavity.
In situ biofilm from healthy individuals was collected for 12 h during day and night, respectively, subjected to fluorescent in situ hybridization and visualized using confocal laser scanning microscopy.
Analysis of the biofilms using stereological methods and digital image analysis revealed a consistent statistically significant difference between both the total number of bacteria and the biovolume in the two 12-h groups (p = 0.012), with the highest accumulation of bacteria during daytime (a factor of 8.8 and 6.1 higher, respectively). Hybridization with probes specific for streptococci and Actinomyces naeslundii indicated a higher proportion of streptococci in biofilms grown during daytime as compared to night-time. No differences could be observed for A. naeslundii. The degree of microbial coverage and the bacterial composition varied considerably between different individuals.
The data provide firm evidence that initial biofilm formation decreases during the night, which may reflect differences in the availability of salivary nutrients. This finding is of significant importance when studying population dynamics during experimental dental biofilm formation.
由于生物膜在口腔疾病中的病因作用,研究牙面初始微生物定植是一个密集研究领域。大多数先前关于体内新生牙菌斑的积累和组成的研究并没有区分白天和夜间形成的生物膜。本研究假设口腔固体表面细菌的积累速度存在昼夜变化。
分别在白天和夜间收集健康个体的原位生物膜 12 小时,进行荧光原位杂交并用共聚焦激光扫描显微镜观察。
使用体视学法和数字图像分析对生物膜进行分析,结果显示两组 12 小时生物膜的总细菌数量和生物体积均存在统计学显著差异(p = 0.012),白天的细菌积累最高(分别高 8.8 和 6.1 倍)。用特异性探针杂交链球菌和内氏放线菌,表明与夜间相比,白天生长的生物膜中链球菌的比例更高。对于内氏放线菌,没有观察到差异。不同个体之间的微生物覆盖率和细菌组成差异很大。
该数据提供了确凿的证据,表明初始生物膜形成在夜间减少,这可能反映了唾液营养物质可用性的差异。在研究实验性牙菌斑形成过程中的群体动态时,这一发现具有重要意义。