Antimicrobial and antibiofilm activity of pleurocidin against cariogenic microorganisms

Dental caries is a common oral bacterial infectious disease of global concern. Prevention and treatment of caries requires control of the dental plaque formed by pathogens such as Streptococcus mutans and Streptococcus sobrinus. Pleurocidin, produced by Pleuronectes americanus, is an antimicrobial peptide that exerts broad-spectrum activity against pathogenic bacteria and fungi. Moreover, pleurocidin shows less hemolysis and is less toxic than other natural peptides. In the present study, we investigated whether pleurocidin is an effective antibiotic peptide against common cariogenic microorganisms and performed a preliminary study of the antimicrobial mechanism. We assayed minimal inhibitory concentration (MIC), minimal bactericide concentration (MBC) and bactericidal kinetics and performed a spot-on-lawn assay. The BioFlux system was used to generate bacterial biofilms under controllable flow. Fluorescence microscopy and confocal laser scanning microscopy (CLSM) were used to analyze and observe biofilms. Scanning electron microscopy was used to observe the bacterial membrane. MIC and MBC results showed that pleurocidin had different antimicrobial activities against the tested oral strains. Although components of saliva could affect antimicrobial activity, pleurocidin dissolved in saliva still showed antimicrobial effects against oral microorganisms. Furthermore, pleurocidin showed a favorable killing effect against BioFlux flow biofilms in vitro. Our findings suggest that pleurocidin has the potential to kill dental biofilms and prevent dental caries.     

The Copenhagen Oral Health Senior Cohort: design, population and dental health

Gerodontology 2010; doi: 10.1111/j.1741‐2358.2010.00383.x The Copenhagen Oral Health Senior Cohort: design, population and dental health Background: In order to study the way old age influence oral health, the Copenhagen Oral Health Senior Cohort (COHS) has been established. Objectives: To describe the design, measurement procedures, and baseline values for COHS including spatial distribution of restorations and dental caries as well as reasons for non‐participation. Materials and methods: Seven hundred and eighty‐three individuals aged 65 years or older, from a total of 1918 invited elderly people, underwent an interview regarding oral health‐related behaviour and a clinical oral examination including measurement of unstimulated whole saliva flow rate. Results: Twelve percent of the COHS was edentulous. The number of dental restorations was higher for women compared to men; however, men had more caries than women. Coronal caries was most frequent on mesial and distal surfaces and on the maxillary incisors and canines; root caries was most frequent on labial surfaces and evenly distributed within the dentition. Only 41% of all invited elderly people accepted the invitation, with old age and poor health being the primary reasons for non‐participation. Conclusion: The baseline values for COHS show that a substantial proportion of the participants had retained a natural dentition and that dental caries was prevalent with the anterior maxillary teeth being most affected.     

Characterization of two operons that encode components of fructose-specific enzyme II of the sugar:phosphotransferase system of Streptococcus mutans

Three genes, designated as fruC, fruD and fruI, were predicted to encode polypeptides homologous to fructose-specific enzyme II (II(Fru)) of the phosphoenolpyruvate-dependent sugar:phosphotransferase system, and were cloned from Streptococcus mutans, the primary etiological agent of human dental caries. The fruC and fruD genes encoded domains BC and domain A of II(Fru), respectively. The fruI gene encoded IICBA(Fru). Northern hybridization and slot blot analysis showed that expression of fruI was inducible by sucrose and fructose, while fruCD were expressed constitutively and at much lower levels. Inactivation of either fruI or fruCD alone, or of both fruCD and fruI, had no major impact on growth on fructose at a concentration of 0.5% (w/v). However, when the strains were grown with 0.2% fructose as the sole carbohydrate source, a significant decrease in the growth rate was seen with the fruCD/fruI double mutants. Assays of sugar:phosphotransferase activity showed that the fruCD/fruI double mutants had roughly 30% of the capacity of the wild-type strain to transport fructose via the phosphoenolpyruvate-dependent sugar:phosphotransferase system. Xylitol toxicity assays indicated that the inducible fructose permease was responsible for xylitol transport.     

A three-species biofilm model for the evaluation of enamel and dentin demineralization

Although Streptococcus mutans biofilms have been useful for evaluating the cariogenic potential of dietary carbohydrates and the effects of fluoride on dental demineralization, a more appropriate biofilm should be developed to demonstrate the influence of other oral bacteria on cariogenic biofilms. This study describes the development and validation of a three-species biofilm model comprising Streptococcus mutans, Actinomyces naeslundii, and Streptococcus gordonii for the evaluation of enamel and dentin demineralization after cariogenic challenges and fluoride exposure. Single- or three-species biofilms were developed on dental substrata for 96?h, and biofilms were exposed to feast and famine episodes. The three-species biofilm model produced a large biomass, mostly comprising S. mutans (41%) and S. gordonii (44%), and produced significant demineralization in the dental substrata, although enamel demineralization was decreased by fluoride treatment. The findings indicate that the three-species biofilm model may be useful for evaluating the cariogenic potential of dietary carbohydrates other than sucrose and determining the effects of fluoride on dental substrata.     

The efficacy of three formulations of Lippia sidoides Cham. essential oil in the reduction of salivary Streptococcus mutans in children with caries: A randomized,double-blind,controlled study

Essential oils of many plants have been previously tested in the treatment of oral diseases and other infections. This study was a randomized, double-blind, in parallel with an active control study, which aimed to evaluate the efficacy of three formulations of the Lippia sidoides Cham. essential oil (LSO) in the reduction of salivary Streptococcus mutans in children with caries. 81 volunteers, aged 6–12 years, both genders, with caries, were recruited to participate in this study, and randomly assigned to either one of five different groups. Each group received topical treatment with either 1.4% LSO toothpaste, 1.4% LSO gel, 0.8% LSO mouthwash, 1% chlorhexidine gel, or 0.12% chlorhexidine mouthwash. A 5-ml volume of each gel was placed inside disposable trays, and applied for 1 min, every 24 h, for 5 consecutive days. The mouthwash groups used 5-ml volume of a mouthwash inside disposable syringes. In the toothpaste group, children brushed their teeth for 1 min, once a day for 5 days. Saliva was collected before and after treatment. MS colonies were counted, isolated and confirmed through biochemical tests. Differences in MS levels measured in different days within the same treatment group was only verified with LSO toothpaste, chlorhexidine gel and chlorhexidine mouthwash. Comparison between groups of LSO mouthwash, toothpaste and gel showed that the toothpaste group expressed significantly lower MS levels than the mouthwash and gel groups at day-30. Chlorhexidine significantly reduced MS levels after 5 days of treatment, but these levels returned to baseline in other periods of the study. LSO toothpaste reduced MS levels after 5 days of treatment, and MS levels remained low and did not return to baseline during subsequent analysis. Hence, LSO toothpaste demonstrated the most long-lasting MS reduction in saliva, whereas other LSO formulations did not effectively reduce MS levels in children with dental caries.     

Number of teeth and risk of root caries

Objective: This study aimed to investigate the association between number of natural teeth and prevalence of root caries. Design: A cross‐sectional design was adopted. A stratified multi‐stage purposive sampling method was used to select a wide range of elderly people from all social classes, educational levels and sexes. Data was collected through interviews and clinical examinations. Setting: Urban area in Chiang Mai‐Thailand. Participants: 549 elderly dentate aged 60‐74 years. Main Outcome Measures: Teeth with decay and filled roots (DF‐T). Results: The prevalence of root caries was 18.2% with a mean DF‐T of 0.58 (S.D. 2.02). Results of multiple logistic regression showed that number of teeth, DMF‐S scores, sex, and socio economic factors were statistically significantly associated with root caries (P<0.05). Adjusted odds ratios demonstrated a 14% increase in the chance of having root caries when the number of teeth increased one unit. Similarly, a 3% increase was observed for each unit increase in the DMF‐S scores (p<0.001). Being male, having more than four years of education and earning more than 1500 baht/month increased the chances of root caries by 76%, 139% and 85% respectively (p<0.001). Recession and age were not statistically significantly associated with root caries. Conclusions: Having more teeth, higher DMF‐S scores, earning more than 1500 baht/month, having more than 4 years of education and being male increased the likelihood of having root caries.     

牙菌斑生物膜的形成与控制

牙菌斑生物膜是附着于牙釉质表面,由复杂的微生物群落构成的一种聚集体。牙菌斑生物膜的形成与生长对口腔健康有着直接或间接的影响,许多研究证实口腔疾病如龋齿和牙周病都与细菌的积累及牙菌斑的形成有关。在牙菌斑生物膜形态建成过程中,牙齿表面最初的定殖菌对生物膜的微生物组成和结构至关重要,这些初级定殖菌决定了后续与之结合形成共生体的微生物种类和数量。不同的微生物组成可能在与生物膜形成相关的口腔病理状况中发挥不同的作用。因此,本文就牙菌斑生物膜的生长及控制进行综述,介绍其微生物的早期定殖和成熟过程、以及通过物理和化学方法对牙菌斑生物膜的控制,以期为了解牙菌斑生物膜的形成机制及相关口腔疾病的预防和治疗提供有价值的参考。     

A lemon myrtle extract inhibits glucosyltransferases activity of Streptococcus mutans

Streptococcus mutans is a bacterium found in human oral biofilms (dental plaques) that is associated with the development of dental caries. Glucosyltransferases (GTFs) are key enzymes involved in dental plaque formation, and compounds that inhibit their activities may prevent dental caries. We developed a screening system for GTF-inhibitory activities, and used it to profile 44 types of herbal tea extracts. Lemon myrtle (Backhousia citriodora) extract exhibited the highest GTF-inhibitory activity, with an IC₅₀ for GTF in solution of 0.14 mg mL^?1. Furthermore, lemon myrtle extracts had the third-highest polyphenol content of all tested extracts, and strongly inhibited S. mutans biofilm. Interestingly, lemon myrtle extracts did not inhibit cell growth.     

广西顶蛳山遗址人骨的龋齿病理观察

顶蛳山遗址位于广西壮族自治区南宁市邕宁区,保存了丰富的新石器时代文化遗存以及大量的古人类骨骼遗存。本文主要从龋齿研究入手,对顶蛳山二、三期文化(8-7 ka BP)中大于15岁的169个体的2737枚恒齿进行了观察。分别统计了龋齿等级和个体和牙齿患龋率以评估人群患龋情况,进而分析顶蛳山遗址人群的口腔健康状况,并探讨了食物构成、饮食行为与社会经济等。统计结果表明,顶蛳山遗址人群的个体和牙齿患龋率都较高。患龋率在两性、葬式、年龄段间都存在差异,女性患龋程度要高于男性;不同葬式之间差异也显著,随着年龄的增长患龋的比例和程度也随之加深。通过与其他8组新石器时代国内遗址人群的个体和牙齿患龋率的对比,我们看到包含顶蛳山在内的三组华南渔猎采集遗址的个体和牙齿患龋率都要高于其他遗址,农业遗址的人群要低于华南遗址的患龋率,而混合经济遗址的患龋率最低。龋齿的出现与人类饮食中的碳水化合物关系密切。据此,我们推测顶蛳山及其他两组华南遗址的高患龋与碳水化合物的摄入关系密切,但这与一般所认为的农业的出现没有联系。三组遗址都是以渔猎采集主导的社会经济形态,并且这时期农业并未传播至华南地区。因此我们的分析表明高患龋率与农业并没有必然联系,而华南地区当时人群所食用的碳水化合物可能源于当地的块茎类和含蔗糖类植物。