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Next-generation sequencing approaches to understanding the oral microbiome. 了解口腔微生物组的新一代测序方法。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449466
E Zaura

Until recently, the focus in dental research has been on studying a small fraction of the oral microbiome-so-called opportunistic pathogens. With the advent of next-generation sequencing (NGS) technologies, researchers now have the tools that allow for profiling of the microbiomes and metagenomes at unprecedented depths. The major advantages of NGS are the high throughput and the fact that specific taxa do not need to be targeted. The relatively low cost and the availability of sequencing facilities have contributed to nearly exponential growth of NGS datasets. The quality and interpretation of the NGS data could be undermined at numerous steps-from sample collection, storage, and DNA extraction to PCR bias, sequencing errors, choice of algorithms for data processing, and statistical analyses. Making sense out of this data deluge is and will be the major challenge. The community analyses based on systems ecology principles will bring us closer to an understanding of the underlying forces that facilitate the stability (or imbalance) of the microbiome. The next logical step will take us beyond the microbiome. The integration of bacterial, viral, fungal "meta-omes" such as the meta-transcriptome, meta-proteome, and meta-metabolome, together with the host as a major co-factor, should be the ultimate goal in unraveling the complexity of the oral interactome.

直到最近,牙科研究的重点一直集中在研究口腔微生物群的一小部分——所谓的机会致病菌。随着下一代测序(NGS)技术的出现,研究人员现在拥有了能够在前所未有的深度上分析微生物组和宏基因组的工具。NGS的主要优点是高通量和不需要针对特定的分类群。相对较低的成本和测序设备的可用性促成了NGS数据集的近乎指数级增长。从样品收集、存储、DNA提取到PCR偏差、测序错误、数据处理算法的选择和统计分析,NGS数据的质量和解释可能在许多步骤中受到影响。从这些数据洪流中寻找意义是一项重大挑战。基于系统生态学原理的群落分析将使我们更接近于理解促进微生物组稳定(或不平衡)的潜在力量。下一个合乎逻辑的步骤将使我们超越微生物组。整合细菌、病毒、真菌“元组”,如元转录组、元蛋白质组和元代谢组,以及作为主要辅助因子的宿主,应该是揭示口腔相互作用复杂性的最终目标。
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引用次数: 73
Clinical efficacy and effects of xylitol wipes on bacterial virulence. 木糖醇湿巾的临床疗效及对细菌毒力的影响。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449835
L Zhan, J D B Featherstone, J Lo, C Krupansky, N Hoang, P DenBesten, T Huynh

The aim of the study was to investigate whether xylitol-wipe use in young children prevented caries by affecting bacterial virulence. In a double-blinded randomized controlled clinical trial, 44 mother-child pairs were randomized to xylitol-wipe or placebo-wipe groups. Salivary mutans streptococci levels were enumerated at baseline, 6 months, and one year. Ten mutans streptococci colonies were isolated and genotyped from each saliva sample. Genotype-colonization stability, xylitol sensitivity, and biofilm formation of these isolates were studied. Despite a significant reduction in new caries at one year in the xylitol-wipe group, no significant differences were found between the two groups in levels of mutans streptococci. Children in the xylitol-wipe group had significantly fewer retained genotypes (p = 0.06) and more transient genotypes of mutans streptococci (p = 0.05) than those in the placebo-wipe group. At one year, there was no significant difference in the prevalence of xylitol-resistant genotypes or in biofilm formation ability of mutans streptococci isolates between the two groups. The mechanism of the caries-preventive effect of xylitol-wipe use may be related to the stability of mutans streptococci colonization. Further studies with genomic characterization methods are needed to determine specific gene(s) that account for the caries-preventive effect of xylitol.

该研究的目的是调查木糖醇湿巾是否通过影响细菌毒力来预防幼儿龋齿。在一项双盲随机对照临床试验中,44对母子被随机分为木糖醇擦拭组和安慰剂擦拭组。在基线、6个月和1年时,对唾液变形链球菌水平进行计数。从每个唾液样本中分离出10个突变链球菌菌落并进行基因分型。研究了这些菌株的基因型定植稳定性、木糖醇敏感性和生物膜形成。尽管木糖醇擦拭组在一年内显著减少了新龋病,但两组之间在变形链球菌水平上没有发现显著差异。与安慰剂组相比,木糖醇擦拭组儿童保留的基因型明显减少(p = 0.06),突变链球菌的瞬时基因型明显增加(p = 0.05)。一年后,两组之间突变链球菌耐木糖醇基因型的患病率或生物膜形成能力没有显著差异。木糖醇湿巾防龋作用的机制可能与变形链球菌定植的稳定性有关。需要进一步研究基因组表征方法来确定木糖醇预防龋齿作用的特定基因。
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引用次数: 15
Caries clinical trial methods for the assessment of oral care products in the 21st century. 21世纪口腔护理产品龋病临床试验评价方法。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449464
R P Ellwood, J Gomez, J Goma, I A Pretty

Traditionally, caries clinical trials of oral care products have focused on the prevention of caries in children and adolescents at the "cavitation" level. Because of a general reduction in caries incidence and the use of positive control comparators, studies have grown both in size and duration to improve statistical power. Currently, they tend to be of 2 to 3 years' duration, with up to 2,000 high-risk individuals per group. During the past decade, there has been a shift in emphasis from a restorative approach to the treatment of dental caries to a therapeutic approach focused on the remineralization of early caries lesions. However, caries clinical trials of oral care products have not often reflected this paradigm change. This manuscript reviews alternative caries clinical trial methods for oral care products. It is concluded that methods focused on the detection and monitoring of enamel caries and root caries, by visual approaches such as ICDAS and instrumental methods such as QLF, Diagnodent, and Electrical Caries Monitors, provide viable alternatives to traditional methods. In particular, such approaches more accurately reflect the modes of action of many therapeutic agents and formulations and may reduce the cost and duration of product innovation.

传统上,口腔护理产品的龋齿临床试验侧重于在“空化”层面预防儿童和青少年龋齿。由于龋齿发生率的普遍降低和阳性对照比较物的使用,研究的规模和持续时间都有所增加,以提高统计能力。目前,它们往往持续2至3年,每组有多达2000名高危个体。在过去的十年中,龋齿治疗的重点从修复方法转向了早期龋齿病变再矿化的治疗方法。然而,口腔护理产品的龋齿临床试验往往没有反映这种范式变化。本文综述了口腔护理产品的替代龋齿临床试验方法。综上所述,通过ICDAS等目视方法和QLF、诊断仪、电龋监测仪等仪器方法对牙釉质龋和牙根龋进行检测和监测,是传统方法的可行替代方法。特别是,这种方法更准确地反映了许多治疗剂和制剂的作用方式,并可能减少产品创新的成本和持续时间。
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引用次数: 28
Remineralization agents - new and effective or just marketing hype? 矿化剂-新的和有效的或只是营销炒作?
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512454295
R J M Lynch, S R Smith

This is a review of the need for better remineralization and of the status of calcium-based remineralizing agents for use in anti-caries toothpastes. Use of fluoride toothpastes has markedly reduced caries. However, the decline may be over or in reverse. There is a limit to what fluoride alone can do; complementary agents are needed. Using plaque as a reservoir for calcium-based agents holds promise. Plaque fluid is already supersaturated with respect to relevant calcium phosphates at neutral pH; extra calcium may lead to surface-blocking and sub-optimal lesion consolidation. However, at cariogenic pH, lesions may be more porous to the ingress of mineral, leading to fuller consolidation, and controlled release of calcium should reduce undersaturation with respect to enamel and accelerate deposition of fluorhydroxyapatite. Clinical data to validate in vitro screening models are scarce. Direct progression to in situ models may often be appropriate. The spectrum of lesion types, from softening to relatively advanced subsurface, and lesion activity should be considered. Far from being 'marketing hype', progress with calcium-based remineralizing agents is both encouraging and scientifically sound. Clinical evidence exists for the efficacy of some agents, but further unequivocal clinical data are needed before these agents might be considered 'effective' when delivered from toothpaste.

本文综述了对更好的再矿化的需求以及钙基再矿化剂在防龋牙膏中的应用现状。使用含氟牙膏可以显著减少龋齿。然而,这种下降可能已经结束,或者正在逆转。氟化物本身的作用是有限的;需要补充剂。利用斑块作为钙基药物的储存库是有希望的。相对于中性pH下的相关磷酸钙,斑块液已经过饱和;过量的钙可能导致表面阻塞和次理想的病变巩固。然而,在致龋的pH值下,病变可能对矿物质的进入更具多孔性,导致更充分的固结,钙的控制释放应减少牙釉质的欠饱和并加速氟羟基磷灰石的沉积。验证体外筛选模型的临床数据很少。直接进展到原位模型可能经常是合适的。应考虑病变类型的谱,从软化到相对较深的地下,以及病变的活动性。钙基再矿化剂的进展不仅令人鼓舞,而且在科学上是合理的,这远远不是“营销炒作”。一些药物的有效性存在临床证据,但在这些药物从牙膏中释放出来被认为是“有效的”之前,还需要进一步明确的临床数据。
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引用次数: 52
The role of matrix metalloproteinases in dental erosion. 基质金属蛋白酶在牙侵蚀中的作用。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512455029
M A R Buzalaf, M T Kato, A R Hannas

This review discusses the role of matrix metalloproteinases (MMPs) in the development of dentin erosion and the protective effects of MMP inhibitors, based on recent evidence from in vitro and in situ studies. MMPs are present in both dentin and saliva and play an important role in dentin erosion progression. Enzymatic removal of the organic matrix by MMPs increases the demineralization process, since the demineralized organic matrix has been shown to hamper ionic diffusion after an acidic challenge. Recent evidence from in vitro and in situ studies has shown a protective role of MMP inhibitors against dentin erosion and erosion plus abrasion. The inhibitors tested were green tea and its active epigallocatechin-gallate (EGCG), ferrous sulfate, and chlorhexidine. They have been tested in dentifrices, solutions, and gels. The latter led to a more pronounced protective effect against dentin erosion and erosion plus abrasion. The protection was long-lasting and could be observed after up to 10 days of severe erosive and erosive-plus-abrasive challenges in situ. Thus, the use of MMP inhibitors has emerged as an important preventive tool against dentin erosion. Clinical studies should be conducted to confirm the results obtained and to give support to the establishment of clinical protocols of use.

本文综述了基质金属蛋白酶(MMPs)在牙本质侵蚀发生中的作用,以及MMP抑制剂的保护作用,基于最近的体外和原位研究证据。MMPs存在于牙本质和唾液中,在牙本质侵蚀过程中起重要作用。通过MMPs酶去除有机基质增加了脱矿过程,因为脱矿的有机基质在酸性挑战后会阻碍离子扩散。最近来自体外和原位研究的证据表明,MMP抑制剂对牙本质侵蚀和侵蚀加磨损具有保护作用。所测试的抑制剂为绿茶及其活性表没食子儿茶素没食子酸酯(EGCG)、硫酸亚铁和氯己定。它们已经在牙膏、溶液和凝胶中进行了测试。后者对牙本质侵蚀和侵蚀加磨损有更明显的保护作用。这种保护是持久的,可以在长达10天的严重侵蚀和侵蚀加磨蚀挑战后观察到。因此,使用MMP抑制剂已成为预防牙本质侵蚀的重要工具。应进行临床研究,以确认所获得的结果,并为建立临床使用方案提供支持。
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引用次数: 81
Probiotics for caries prevention and control. 预防和控制龋齿的益生菌。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449465
S Twetman, M K Keller

Modulation of the microbiota for restoring and maintaining health is a growing issue in medical science. A search for relevant clinical trials on the use of probiotic bacteria as a potential and clinically applicable anti-caries measure was performed. According to predetermined criteria, papers were selected and key data on study design, sample size, intervention, duration, and results were extracted. Two animal and 19 human studies were retrieved. Most studies were short-term and restricted to microbiological endpoints, and only 3 human studies reported a caries endpoint. A high degree of heterogeneity among the included investigations hampered the analysis. Significant reductions of mutans streptococci in saliva or plaque following daily intake of probiotic lactobacilli or bifidobacteria were reported in 12 out of 19 papers, whereas 3 reported an increase of lactobacilli. Three caries trials in preschool children and the elderly demonstrated prevented fractions of between 21% and 75% following regular intakes of milk supplemented with L. rhamnosus. No adverse effects or potential risks were reported. The currently available literature does not exclude the possibility that probiotic bacteria can interfere with the oral biofilm, but any clinical recommendation would be premature. Large-scale clinical studies with orally derived specific anti-caries candidates are still lacking.

调节微生物群以恢复和维持健康是医学中一个日益重要的问题。研究了益生菌作为一种潜在的、临床适用的抗龋措施的相关临床试验。按照预先确定的标准选择论文,提取研究设计、样本量、干预、持续时间和结果等关键数据。检索了两项动物研究和19项人类研究。大多数研究是短期的,仅限于微生物终点,只有3个人类研究报告了龋齿终点。纳入调查的高度异质性阻碍了分析。19篇论文中有12篇报道了每日摄入益生菌乳酸杆菌或双歧杆菌后唾液或牙菌斑中变形链球菌的显著减少,而3篇报道了乳酸杆菌的增加。在学龄前儿童和老年人中进行的三项龋齿试验表明,定期摄入含有鼠李糖乳杆菌的牛奶后,预防龋齿的比例在21%至75%之间。无不良反应或潜在风险报告。目前可获得的文献并不排除益生菌干扰口腔生物膜的可能性,但任何临床推荐都为时过早。大规模的临床研究仍然缺乏口服衍生的特异性抗龋候选物。
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引用次数: 103
Light therapy: complementary antibacterial treatment of oral biofilm. 光疗:口腔生物膜的辅助抗菌治疗。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449469
O Feuerstein

Conventional antibacterial treatment fails to eradicate biofilms associated with common infections of the oral cavity. Unlike chemical agents, which are less effective than anticipated, owing to diffusion limitations in biofilms, light is more effective on bacteria in biofilm than in suspension. Effectiveness depends also on the type and parameters of the light. We tested the phototoxic effects of non-coherent blue light (wavelengths, 400-500 nm) and CO(2) laser (wavelength, 10.6 μm), which have different mechanisms of action on the oral bacterium Streptoccocus mutans, in biofilm and on tooth enamel. Exposure of S. mutans in biofilm to blue light had a delayed effect on bacterial viability throughout the biofilm and a sustained antibacterial effect on biofilm newly formed by previously irradiated bacteria. A synergistic antibacterial effect between blue light and H(2)O(2) may enhance the phototoxic effect, which involves a photochemical mechanism mediated by reactive oxygen species (ROS) formation. The effect of CO(2) laser irradiation on the viability of S. mutans in biofilm on enamel samples appeared to be higher in the deep layers, due to heating of the enamel surface by the absorbed energy. Biofilms do not interfere with the chemical changes resulting from irradiation, which may increase the enamel's resistance to acid attack.

传统的抗菌治疗不能根除与口腔常见感染相关的生物膜。由于生物膜中的扩散限制,化学试剂的效果不如预期,而光对生物膜中的细菌比悬浮液更有效。效果还取决于光的类型和参数。研究了波长400 ~ 500 nm的非相干蓝光和波长10.6 μm的CO(2)激光对口腔变形链球菌、生物膜和牙釉质的光毒性作用。生物膜中的变形链球菌暴露在蓝光下对整个生物膜的细菌活力有延迟影响,对先前照射过的细菌新形成的生物膜有持续的抗菌作用。蓝光和H(2)O(2)之间的协同抑菌作用可能会增强光毒性作用,这涉及活性氧(ROS)形成介导的光化学机制。CO(2)激光照射对牙釉质样品生物膜中变形链球菌活力的影响在深层表现得更高,这是由于吸收的能量加热了牙釉质表面。生物膜不会干扰照射引起的化学变化,而化学变化可能会增加牙釉质对酸的抵抗力。
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引用次数: 21
Delivery of active agents from chewing gum for improved remineralization. 从口香糖中输送活性剂以改善再矿化。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512452886
M W J Dodds, D Chidichimo, M S Haas

Most surrogate measures of caries were developed to test products containing fluoride, typically at relatively high and closely controlled oral concentrations. However, since the primary mechanism for the remineralization of early enamel caries lesions by chewing gum is through stimulation of saliva, delivering Ca and Pi to the demineralized enamel lesion, established methods may lack the sensitivity to detect the additional benefit of an active agent without the strong remineralizing potential of fluoride. Issues related to the release of active agents from the gum matrix, dilution in the saliva, and limited oral retention time, along with taste, safety, regulatory, and cost concerns, impose further limitations. This paper reviews the efficacy of some active agents used in chewing gum for improved remineralization and includes results from in situ testing of calcium-containing gums, including calcium lactate, tetracalcium phosphate/dicalcium phosphate anhydrous, calcium citrate/encapsulated phosphate, and a calcium lactate/sodium phosphate blend. Despite promising in vitro data from these agents, they did not provide consistently superior results from in situ testing. There is a need to develop better predictive in vitro models for chewing gum, as well as improved sensitivity of in situ models to discriminate relatively small amounts of remineralization against a background of high biological variability.

大多数龋齿的替代措施都是用来测试含氟产品的,通常是相对较高且严格控制的口服浓度。然而,由于口香糖对早期牙釉质龋损伤的再矿化的主要机制是通过刺激唾液,将Ca和Pi传递到脱矿的牙釉质病变,现有的方法可能缺乏灵敏度,无法检测活性剂的额外益处,而没有氟化物的强大再矿化潜力。与从牙龈基质中释放活性剂、唾液中的稀释、有限的口腔保留时间以及口味、安全性、监管和成本方面的问题有关的问题进一步限制了该技术的发展。本文综述了口香糖中用于改善再矿化的一些活性剂的功效,并包括对含钙口香糖的原位测试结果,包括乳酸钙、磷酸四钙/无水磷酸二钙、柠檬酸钙/包封磷酸,以及乳酸钙/磷酸钠混合物。尽管这些药物的体外数据很有希望,但它们并没有在原位测试中提供一致的优越结果。有必要开发更好的预测口香糖的体外模型,以及提高原位模型在高生物变异性背景下区分相对少量再矿化的敏感性。
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引用次数: 30
Clinical evidence for polyol efficacy. 多元醇疗效的临床证据。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512449467
P Milgrom, E M Söderling, S Nelson, D L Chi, Y Nakai

Xylitol is a safe dental caries preventive when incorporated into chewing gum or confections used habitually. The goal of this paper is to identify and assess the work on xylitol and other polyols and dental caries since 2008. Xylitol is effective when used by the mother prenatally or after delivery to prevent mutans transmission and subsequent dental caries in the offspring. One new completed trial confirmed that children of mothers who used xylitol lozenges after delivery had less dental caries than a comparison group. A similar study confirmed that the use of xylitol gum by the mother either prevented or postponed MS transmission to the offspring. Xylitol use among schoolchildren delivered via a gummy bear confection reduced S. mutans levels, but a once per day use of xylitol-containing toothpaste did not. Randomized trials, with caries outcomes, assessing xylitol-containing lozenges in adults and xylitol-containing gummy bears in children will release results in the coming year. Other studies are ongoing but are not systematic and will fail to answer important questions about how xylitol, or other polyols, can address the global dental caries problem.

木糖醇是一种安全的龋齿预防剂,当它被加入到口香糖或经常使用的糖果中时。本文的目的是对2008年以来木糖醇等多元醇与龋病的研究进展进行综述和评价。木糖醇是有效的,当使用由母亲产前或分娩后,以防止变异传播和随后的龋齿的后代。一项新完成的试验证实,在分娩后使用木糖醇含片的母亲的孩子比对照组的孩子患龋齿的几率要低。一项类似的研究证实,母亲使用木糖醇胶可以防止或推迟MS传给后代。在小学生中,通过小熊软糖糖果使用木糖醇降低了变形链球菌的水平,但每天使用一次含木糖醇的牙膏却没有。评估成人含木糖醇含片和儿童含木糖醇小熊软糖效果的随机试验将在明年公布结果。其他研究正在进行中,但不是系统的,并且无法回答关于木糖醇或其他多元醇如何解决全球龋齿问题的重要问题。
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引用次数: 44
Remineralization models. 补充矿质模型。
Q1 Medicine Pub Date : 2012-09-01 DOI: 10.1177/0022034512453845
N J Cochrane, D T Zero, E C Reynolds

During the ICNARA 2 conference, a workshop was held on remineralization models. The group considered the role of remineralization models, whether there was one ideal in situ model design, what essential features should be incorporated into an in situ model, other alternative models, and what new methods for measuring remineralization were on the horizon. This paper summarizes the discussion. In situ and other caries models can be used as a surrogate for caries clinical trials but only when data exist to validate the model. In situ model design should be flexible to allow for investigation of different aspects of the caries process; however, several essential features were identified that should be incorporated into the study design. A range of other caries models was discussed, including the study of non-cavitated lesions, lesions post-orthodontic therapy, plaque retention models to form more standardized lesions, and the study of root caries lesions. Numerous new methods for quantifying remineralization were discussed, but it was considered that these require validation before they can be used in clinical trials.

在ICNARA第二次会议期间,举办了一个关于再矿化模型的讲习班。该小组审议了再矿化模型的作用,是否存在一种理想的原位模型设计,应将哪些基本特征纳入原位模型,其他替代模型,以及即将出现哪些测量再矿化的新方法。本文对讨论进行了总结。原位龋模型和其他龋模型可以作为龋临床试验的替代品,但只有在有数据验证模型的情况下才能使用。原地模型设计应灵活,以便对龋蚀过程的不同方面进行调查;然而,确定了几个基本特征,应纳入研究设计。讨论了一系列其他龋齿模型,包括非空化病变的研究、病变正畸后治疗、形成更标准化病变的牙菌斑保留模型以及牙根龋齿病变的研究。讨论了许多量化再矿化的新方法,但认为这些方法在用于临床试验之前需要验证。
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引用次数: 17
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Advances in Dental Research
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