Cao Li, Zhang Zhen-Zhen, Xu Shuang-Bo, Ma Ming, Wei Xin
Jiangsu Key Laboratory of Oral Diseases, Department of Operative Dentistry and Endodontics, School of Stomatology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
J Biomed Res. 2017 Jul 13;31(4):333-343. doi: 10.7555/JBR.31.20150151.
Streptococcus mutans is a primary etiological agent of dental caries. Farnesol, as a potential antimicrobial agent, inhibits the development ofS. mutans biofilm. In this study, we hypothesized that farnesol inhibits caries development in vitro and interferes with biofilm formation by regulating virulence-associated gene expression. The inhibitory effects of farnesol to S. mutans biofilms on enamel surfaces were investigated by determining micro-hardness and calcium measurements. Additionally, the morphological changes ofS. mutans biofilms were compared using field emission scanning electron microscopy and confocal laser scanning microscopy, and the vitality and oxygen sensitivity ofS. mutans biofilms were compared using MTT assays. To investigate the molecular mechanisms of farnesol's effects, expressions of possible target genesluxS, brpA, ffh, recA, nth, and smx were analyzed using reverse-transcription polymerase chain reaction (PCR) and quantitative PCR. Farnesol-treated groups exhibited significantly higher micro-hardness on the enamel surface and lower calcium concentration of the supernatants as compared to the-untreated control. Microscopy revealed that a thinner film with less extracellular matrix formed in the farnesol-treated groups. As compared to the-untreated control, farnesol inhibited biofilm formation by 26.4% with 500 µmol/L and by 37.1% with 1,000 µmol/L (P<0.05). Last, decreased transcription levels of luxS, brpA, ffh, recA, nth, and smx genes were expressed in farnesol-treated biofilms. In vitrofarnesol inhibits caries development and S. mutans biofilm formation. The regulation of luxS, brpA, ffh, recA, nth, and smx genes may contribute to the inhibitory effects of farnesol.
变形链球菌是龋齿的主要病原体。法尼醇作为一种潜在的抗菌剂,可抑制变形链球菌生物膜的形成。在本研究中,我们假设法尼醇在体外可抑制龋齿的发展,并通过调节毒力相关基因的表达来干扰生物膜的形成。通过测定显微硬度和钙含量,研究了法尼醇对牙釉质表面变形链球菌生物膜的抑制作用。此外,使用场发射扫描电子显微镜和共聚焦激光扫描显微镜比较了变形链球菌生物膜的形态变化,并使用MTT法比较了变形链球菌生物膜的活力和氧敏感性。为了研究法尼醇作用的分子机制,使用逆转录聚合酶链反应(PCR)和定量PCR分析了可能的靶基因luxS、brpA、ffh、recA、nth和smx的表达。与未处理的对照组相比,法尼醇处理组的牙釉质表面显微硬度显著更高,上清液中的钙浓度更低。显微镜检查显示,法尼醇处理组形成的生物膜更薄,细胞外基质更少。与未处理的对照组相比,500 μmol/L的法尼醇抑制生物膜形成26.4%,1000 μmol/L的法尼醇抑制生物膜形成37.1%(P<0.05)。最后,在法尼醇处理的生物膜中,luxS、brpA、ffh、recA、nth和smx基因的转录水平降低。体外实验表明,法尼醇可抑制龋齿的发展和变形链球菌生物膜的形成。luxS、brpA、ffh、recA、nth和smx基因的调节可能有助于法尼醇的抑制作用。