Djais Ariadna A, Putri Nadhifa, Rahmania Putri Andin, Angky Soekanto Sri
Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jalan Salemba Raya No. 4, Jakarta Pusat, Jakarta, 10430, Indonesia.
Saudi Dent J. 2020 Mar;32(3):129-134. doi: 10.1016/j.sdentj.2019.08.003. Epub 2019 Aug 27.
Dental caries is a major and chronic dental public health problem, which can usually be prevented by regular oral hygiene. The most common oral hygiene practice is brushing teeth with a dentifrice. Propolis has emerged as a promising anti-cariogenic agent, which is considered to be a good oral antiseptic for prevention of caries. Several studies have shown that the use of C has an influence in the growth of oral biofilms. There are several standard methods used to count bacterial colonies, such as crystal violet and CFU Count assays. OpenCFU method is a technique that can be used to calculate biofilm colonies more faster, precisely, and accurately.
To compare several methods for evaluating the number of biofilm colonies formed with exposure to a standard dentifrice and propolis.
Biofilm assays were carried out on 96-well microplates. Reference strains of oral species ( ATCC 25175 and ATCC 10566) and yeast ( ATCC 10231) were inoculated into wells, and 200 µL of standard and propolis dentifrice solution were added to each well and incubated for 18 h at 37 °C. Bacteria and yeast were then sub-cultured on respective media and the colony-forming units (CFU) were counted manually. The other wells were stained by crystal violet and incubated for 15 min, followed by observation using an inverted microscope and evaluated using crystal violet analysis and the OpenCFU automated method.
The numbers of CFUs determined for all strains were similar in the standard-dentifrice group and propolis-dentifrice group, and were similar among the three methods: crystal violet staining, manual CFU count, and OpenCFU analysis.
OpenCFU analysis can be reliably used as a rapid and a more practical method to analyse the growth of oral microorganism biofilms. However, high digital image quality is required to provide an accurate analysis for colony counting.
龋齿是一个主要的慢性口腔公共卫生问题，通常可通过定期口腔卫生措施预防。最常见的口腔卫生做法是使用牙膏刷牙。蜂胶已成为一种有前景的抗龋齿剂，被认为是预防龋齿的良好口腔消毒剂。多项研究表明，使用蜂胶对口腔生物膜的生长有影响。有几种用于计数细菌菌落的标准方法，如结晶紫和CFU计数测定法。OpenCFU方法是一种可用于更快、更精确和准确地计算生物膜菌落的技术。
比较几种评估暴露于标准牙膏和蜂胶后形成的生物膜菌落数量的方法。
在96孔微孔板上进行生物膜测定。将口腔菌种（ATCC 25175和ATCC 10566）和酵母（ATCC 10231）的参考菌株接种到孔中，向每个孔中加入200µL标准牙膏和蜂胶牙膏溶液，并在37°C下孵育18小时。然后将细菌和酵母分别接种到各自的培养基上，手动计数菌落形成单位（CFU）。其他孔用结晶紫染色并孵育15分钟，然后使用倒置显微镜观察，并使用结晶紫分析和OpenCFU自动化方法进行评估。
在标准牙膏组和蜂胶牙膏组中，所有菌株的CFU数量相似，并且在三种方法（结晶紫染色、手动CFU计数和OpenCFU分析）之间也相似。
OpenCFU分析可可靠地用作分析口腔微生物生物膜生长的快速且更实用的方法。然而，需要高质量的数字图像才能为菌落计数提供准确的分析。
