Influences of naturally occurring agents in combination with fluoride on gene expression and structural organization of Streptococcus mutans in biofilms.
Influences of naturally occurring agents in combination with fluoride on gene expression and structural organization of Streptococcus mutans in biofilms.
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DOI：
10.1186/1471-2180-9-228
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发表时间：
2009-10-28
期刊：
影响因子：
4.2
通讯作者：
Koo H
中科院分区：
文献类型：
--
作者：
Jeon JG;Klein MI;Xiao J;Gregoire S;Rosalen PL;Koo H
特定的生物活性黄酮类化合物和萜类化合物与氟化物的结合可以通过同时影响变形链球菌胞外多糖（EPS）的合成和酸的产生来调节致龋生物膜的发展，从而增强氟化物在体内的防龋效果。在本研究中，我们进一步研究了杨梅素（黄酮类化合物）、tt-法尼醇（萜类化合物）和氟化物的组合的生物学作用是否会影响S.使用实时PCR和共聚焦荧光显微镜观察生物膜内的变形菌及其结构组织。在生物膜形成期间每天两次处理（一分钟暴露）影响了S.在生物膜发展的早期（49-h）和晚期（97-h）阶段均存在变形杆菌。用试剂组合处理的生物膜显示出较低的gtfB和gtfD（与胞外多糖合成相关）和aguD（与S.变形杆菌酸耐受性）比用载体对照处理的那些（p < 0.05）。此外，药物联合处理显著影响了S.通过减少生物体积（生物量）和EPS和细菌细胞在生物膜深度上的比例，特别是在中间层和外层中，可以减少变形杆菌生物膜（相对于媒介物对照，p < 0.05）。与载体处理的生物膜相比，用试剂的组合处理的生物膜也较少产酸，并且具有减少量的细胞外不溶性葡聚糖和细胞内多糖（p < 0.05）。数据表明，天然存在的因子与氟化物的组合有效地破坏了特定毒力基因的表达、结构组织和S.变形杆菌生物膜，这可以解释我们的化疗方法的增强的防龋效果。
The association of specific bioactive flavonoids and terpenoids with fluoride can modulate the development of cariogenic biofilms by simultaneously affecting the synthesis of exopolysaccharides (EPS) and acid production by Streptococcus mutans, which enhanced the cariostatic effectiveness of fluoride in vivo. In the present study, we further investigated whether the biological actions of combinations of myricetin (flavonoid), tt-farnesol (terpenoid) and fluoride can influence the expression of specific genes of S. mutans within biofilms and their structural organization using real-time PCR and confocal fluorescence microscopy. Twice-daily treatment (one-minute exposure) during biofilm formation affected the gene expression by S. mutans both at early (49-h) and later (97-h) stages of biofilm development. Biofilms treated with combination of agents displayed lower mRNA levels for gtfB and gtfD (associated with exopolysaccharides synthesis) and aguD (associated with S. mutans acid tolerance) than those treated with vehicle-control (p < 0.05). Furthermore, treatment with combination of agents markedly affected the structure-architecture of S. mutans biofilms by reducing the biovolume (biomass) and proportions of both EPS and bacterial cells across the biofilm depth, especially in the middle and outer layers (vs. vehicle-control, p < 0.05). The biofilms treated with combination of agents were also less acidogenic, and had reduced amounts of extracellular insoluble glucans and intracellular polysaccharides than vehicle-treated biofilms (p < 0.05). The data show that the combination of naturally-occurring agents with fluoride effectively disrupted the expression of specific virulence genes, structural organization and accumulation of S. mutans biofilms, which may explain the enhanced cariostatic effect of our chemotherapeutic approach.
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