Cranberry Flavonoids Modulate Cariogenic Properties of Mixed-Species Biofilm through Exopolysaccharides-Matrix Disruption.
Cranberry Flavonoids Modulate Cariogenic Properties of Mixed-Species Biofilm through Exopolysaccharides-Matrix Disruption.
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DOI：
10.1371/journal.pone.0145844
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Koo H
中科院分区：
文献类型：
--
作者：
Kim D;Hwang G;Liu Y;Wang Y;Singh AP;Vorsa N;Koo H
变形链球菌葡糖基转移酶（Gtfs）产生的胞外多糖（EPS）是与致龋生物膜形成相关的重要毒力因子。EPS形成生物膜基质-支架的核心，提供机械稳定性，同时促进局部酸性微环境的产生。蔓越莓类黄酮，如A型原花青素（PAC）和杨梅素，已被证明可以抑制Gtfs和EPS介导的细菌粘附的活性，而不会杀死生物体。在这里，我们研究了蔓越莓类黄酮的组合是否会破坏EPS的积累和S。在致龋条件下使用混合物种生物膜模型观察变形杆菌的存活。我们还评估了蔓越莓类黄酮对机械稳定性和生物膜-磷灰石界面处的原位pH的影响。使用PAC寡聚物（100-300 μM）与杨梅素（2 mM）每日两次的优化组合的局部应用来模拟临床上经历的治疗方案。蔓越莓黄酮处理有效降低了不溶性EPS含量（与溶剂对照相比降低>80%; p<0.001），同时抑制了S.混合物种生物膜内的变异体生长。结果，蔓越莓处理的生物膜的3D结构受到严重损害，显示出有缺陷的EPS基质和未能在唾液涂覆的羟基磷灰石（sHA）表面上形成小菌落。此外，蔓越莓类黄酮的局部应用显著削弱了生物膜的机械稳定性;在暴露于0.449 N/m2的剪切应力后，从sHA表面去除了近90%的生物膜（相对于在载体处理的生物膜中去除36%）。重要的是，蔓越莓处理的生物膜中的原位pH测量显示生物膜-磷灰石界面处的pH值（5.2 ± 0.1）显著高于载体处理的生物膜（4.6 ± 0.1）。总而言之，这些数据提供了重要的见解，蔓越莓黄酮类化合物治疗如何调节毒力特性，通过破坏与致龋生物膜发展相关的生物化学和生态变化，这可能会导致新的替代或抑制性抗龋膜/抗龋化疗制剂。
The exopolysaccharides (EPS) produced by Streptococcus mutans-derived glucosyltransferases (Gtfs) are essential virulence factors associated with the initiation of cariogenic biofilms. EPS forms the core of the biofilm matrix-scaffold, providing mechanical stability while facilitating the creation of localized acidic microenvironments. Cranberry flavonoids, such as A-type proanthocyanidins (PACs) and myricetin, have been shown to inhibit the activity of Gtfs and EPS-mediated bacterial adhesion without killing the organisms. Here, we investigated whether a combination of cranberry flavonoids disrupts EPS accumulation and S. mutans survival using a mixed-species biofilm model under cariogenic conditions. We also assessed the impact of cranberry flavonoids on mechanical stability and the in situ pH at the biofilm-apatite interface. Topical application of an optimized combination of PACs oligomers (100–300 μM) with myricetin (2 mM) twice daily was used to simulate treatment regimen experienced clinically. Treatments with cranberry flavonoids effectively reduced the insoluble EPS content (>80% reduction vs. vehicle-control; p<0.001), while hindering S. mutans outgrowth within mixed-species biofilms. As a result, the 3D architecture of cranberry-treated biofilms was severely compromised, showing a defective EPS-matrix and failure to develop microcolonies on the saliva-coated hydroxyapatite (sHA) surface. Furthermore, topical applications of cranberry flavonoids significantly weaken the mechanical stability of the biofilms; nearly 90% of the biofilm was removed from sHA surface after exposure to a shear stress of 0.449 N/m2 (vs. 36% removal in vehicle-treated biofilms). Importantly, in situ pH measurements in cranberry-treated biofilms showed significantly higher pH values (5.2 ± 0.1) at the biofilm-apatite interface vs. vehicle-treated biofilms (4.6 ± 0.1). Altogether, the data provide important insights on how cranberry flavonoids treatments modulate virulence properties by disrupting the biochemical and ecological changes associated with cariogenic biofilm development, which could lead to new alternative or adjunctive antibiofilm/anticaries chemotherapeutic formulations.