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Quaternary ammonium-induced multidrug tolerant Streptococcus mutans persisters elevate cariogenic virulence in vitro

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International Journal of Oral Science 2017年第4期9卷 224-231页

作者： Ya-Ling Jiang Wei Qiu Xue-Dong Zhou Hao Li Jun-Zhuo Lu Hockin HK Xu Xian Peng Ming-Yun Li Ming-Ye Feng Lei Cheng Biao Ren State Key Laboratory of Oral Diseases&National Clinical Research Center for Oral Diseases&Department of Operative Dentistry and Endodontics West China Hospital of StomatologySichuan UniversityChengduChina Biomaterials&Tissue Engineering Division Department of EndodonticsProsthodontics and Operative DentistryUniversity of Maryland Dental SchoolBaltimoreUSA

dental caries are the most prevalent chronic infections in the oral cavity, and Streptococcus mutans acts as the main cariogenic bacterial species. Antibacterial quaternary ammonium compounds （QAs） have been developed to preveFnt or treat dental caries. However, there is no report on the tolerance of S. mutans to QAs. In this study, we investigated the development of S. mutans persistence induced by a novel dental caries defensive agent, dimethylaminododecyl methacrylate （DMADDM）, for the first time. Typical biphasic killing kinetics for persisters were observed in both S. mutans planktonic and biofilm cultures challenged by DMADDM at concentrations of 20 and 200 ***- z respectively. The persisters tolerated six other antibiotics with different antibacterial mechanisms, while only daptomycin and vancomycin could slightly reduce the persister numbers in planktonic cultures. The distribution of persisters in DMADDM-treated biofilms was similar to that in the untreated control, except that the total biomass and biofilm height were significantly reduced. A higher exopolysaccharides （EPS）：bacteria ratio was observed in DMADDM-treated biofilms. Persisters in biofilms significantly upregulated gtf gene expression, indicating an increase in the bacteria＇s ability to produce EPS and an elevated capability of cariogenic virulence. Carbon source metabolism was significantly reduced, as related metabolic genes were all downregulated in persisters. Concentrations of 0.1 mM, 1 mM and 10 mM of extra glucose significantly reduced the number of persisters both in planktonic and biofilm conditions. The formation of non- inheritable and multidrug tolerant persisters induced by DMADDM suggested that drug tolerance and new persistent eradication strategies should be considered for oral antibacterial agents.

关键词： antibacterial quaternary ammoniums dental caries persistence multidrug tolerance Streptococcus mutans biofilms

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Biofilm three-dimensional architecture influences in situ pH distribution pattern on the human enamel surface

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International Journal of Oral Science 2017年第2期9卷 74-79页

作者： Jin Xiao Anderson T Hara Dongyeop Kim Domenick T Zero Hyun Koo Geelsu Hwang Eastman Institute for Oral Health University of Rochester Medical Center Department of Cariology Operative Dentistry and Dental Public HealthOral Health Research InstituteIndiana University School of Dentistry Biofilm Research Laboratories Levy Center for Oral HealthDepartment of Orthodontics and Divisions of Pediatric Dentistry & Community Oral HealthSchool of Dental MedicineUniversity of Pennsylvania

To investigate how the biofilm three-dimensional(3D) architecture influences in situ pH distribution patterns on the enamel surface. Biofilms were formed on human tooth enamel in the presence of 1% sucrose or 0.5% glucose plus 0.5% fructose. At specific time points, biofilms were exposed to a neutral pH buffer to mimic the buffering of saliva and subsequently pulsed with 1% glucose to induce re-acidification. Simultaneous 3D pH mapping and architecture of intact biofilms was performed using two-photon confocal microscopy. The enamel surface and mineral content characteristics were examined successively via optical profilometry and microradiography analyses. Sucrose-mediated biofilm formation created spatial heterogeneities manifested by complex networks of bacterial clusters(microcolonies). Acidic regions(pH<5.5) were found only in the interior of microcolonies,which impedes rapid neutralization(taking more than 120 min for neutralization). Glucose exposure rapidly re-created the acidic niches, indicating formation of diffusion barriers associated with microcolonies structure. Enamel demineralization(white spots),rougher surface, deeper lesion and more mineral loss appeared to be associated with the localization of these bacterial clusters at the biofilm-enamel interface. Similar 3D architecture was observed in plaque-biofilms formed in vivo in the presence of sucrose. The formation of complex 3D architectures creates spatially heterogeneous acidic microenvironments in close proximity of enamel surface, which might correlate with the localized pattern of the onset of carious lesions(white spot like) on teeth.

关键词： biofilms demineralization dental caries exopolysaccharides microcolonies pH microenvironments plaque Streptococcus mutans

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In silico identification of potential inhibitors targeting Streptococcus mutans sortase A

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International Journal of Oral Science 2017年第1期9卷 53-62页

作者： Hao Luo Dan-Feng Liang Min-Yue Bao Rong Sun Yuan-Yuan Li Jian-Zong Li Xin Wang Kai-Min Lu Jin-Ku Bao School of Life Sciences and Key Laboratory of Ministry of Education for Bio-Resources and Bio-Environment Sichuan University State Key Laboratory of Biotherapy/Collaborative Innovation Centre for Biotherapy West China HospitalSichuan University State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesWest China Hospital of StomatologySichuan University

dental caries is one of the most common chronic diseases and is caused by acid fermentation of bacteria adhered to the teeth. Streptococcus mutans （S. mutans） utilizes sortase A （SrtA） to anchor surface proteins to the cell wall and forms a biofilm to facilitate its adhesion to the tooth surface. Some plant natural products, especially several flavonoids, are effective inhibitors of SrtA. However, given the limited number of inhibitors and the development of drug resistance, the discovery of new inhibitors is urgent. Here, the high-throughput virtual screening approach was performed to identify new potential inhibitors of S. mutans SrtA. Two libraries were used for screening, and nine compounds that had the lowest scores were chosen for further molecular dynamics simulation, binding free energy analysis and absorption, distribution, metabolism, excretion and toxicity （ADMET） properties analysis. The results revealed that several similar compounds composed of benzofuran, thiadiazole and pyrrole, which exhibited good affinities and appropriate pharmacokinetic In addition, the carbonyl of these compounds can have a strategy for microbial infection disease therapy. parameters, were potential inhibitors to impede the catalysis of SrtA. key role in the inhibition mechanism. These findings can provide a new

关键词： dental caries molecular dynamics simulation molecular docking potential inhibitors sortase A Streptococcus mutans

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caries Patterns in the Primary Dentition: Cluster Analysis of a Sample of 5,169 Arizona Children 5-59 Months of Age

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International Journal of Oral Science 2009年第4期1卷 189-195页

作者： Walter J. Psoter David G. Pendrys Douglas E. Morse He-ping Zhang Susan T. Mayne Department of Epidemiology ＆ Health Promotion College of Dentistry New York University New York USA Department of Oral Health and Diagnostic Sciences School of Dental Medicine University Of Connecticut Health CenterFarmington USA Department of Epidemiology and Public Health School of Medicine Yale University New Haven USA

Aim Cluster analysis was conducted on data from 5,169 United States （U.S.） Arizona children, age＇s 5-59-months with the goal of delineating patterns of caries in the primary dentition of pre-school children without a priori pattern definitions. Methodology Cluster analyses were conducted using all data for children ages 0-4 years in aggregate： 1） for all subjects, and 2） for subjects without crowned restored teeth. Each of these two sets of analyses consisted of 8 differently specified cluster analyses as a validation procedure. Results The caries patterns identified from the clustering analysis are： 1） smooth surfaces （other than the maxillary incisor）, 2） maxillary incisor, 3） occlusal surfaces of first molars, and 4） pit and fissure surfaces of second molars. Conclusion The cluster analysis findings were consistent with results produced by multidimensional scaling. These cross-validated patterns may represent resulting disease conditions from different risks or the timing of various risk factor exposures. As such, the patterns may be useful case definitions for caries risk factor investigations in children under 60 months of age.

关键词： dental caries cluster analysis multidimensional scaling early childhood caries (ECC) caries patterns

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Remineralization of initial enamel caries in vitro using a novel peptide based on amelogenin

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Frontiers of Materials Science 2015年第3期9卷 293-302页

作者： Danxue LI Xueping LV Huanxin TU Xuedong ZHOU Haiyang YU Linglin ZHANG State Key Laboratory of Oral Diseases West China Hospital of Stomatology Sichuan University Chengdu 610041 China

dental caries is the most common oral disease with high incidence, widely spread and can seriously affect the health of oral cavity and the whole body. Current caries prevention measures such as fluoride treatment, antimicrobial agents, and traditional Chinese herbal, have limitations to some extent. Here we design and synthesize a novel peptide based on the amelogenin, and assess its ability to promote the remineralization of initial enamel caries lesions. We used enamel blocks to form initial lesions, and then subjected to 12-day pH cycling in the presence of peptide, NaF and HEPES buffer. Enamel treated with peptide or NaF had shallower, narrower lesions, thicker remineralized surfaces and less mineral loss than enamel treated with HEPES. This peptide can promote the remineralization of initial enamel caries and inhibit the progress of caries. It is a promising anti-caries agent with various research prospects and practical application value.

关键词： dental caries peptide remineralization

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Chemical composition of Galla chinensis extract and the effect of its main component(s) on the prevention of enamel demineralization in vitro

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International Journal of Oral Science 2012年第3期4卷 146-151页

作者： Xue-Lian Huang Ming-Dong Liu Ji-Yao Li Xue-Dong Zhou Jacob M ten Cate State Key Laboratory of Oral Diseases Sichuan UniversityChengduChina West China Hospital of Stomatology Sichuan UniversityChengduChina Department of Cariology Endodontology Pedodontology Academic Centre for Dentistry Amsterdam(ACTA)University of Amsterdam and Free University AmsterdamAmsterdamThe Netherlands Department of Analytical Toxicology West China School of Preclinical and Forensic MedicineSichuan UniversityChengduChina

To determine the chemical composition of Galla chinensisextract （GCE） by several analysis techniques and to compare the efficacy of GCE and its main component（s） in inhibition of enamel demineralization, for the development of future anticaries agents, main organic composition of GCE was qualitatively determined by liquid chromatography-time of flight-mass spectrometry （LC-TOF-MS） and quantified by high-performance liquid chromatography-diode array detector （HPLC-DAD）. Inorganic ions were tested by inductively coupled plasma-atomic emission spectroscopy and F was especially measured by ion chromatography. Then, bovine enamel blocks were randomly divided into four treatment groups and were subjected to a pH-cycling regime for 12 times. Each cycle included 5-min applications with one of four treatments： 4g.L-1 GCE solution, 4g.L-1 gallic acid （GA） solution, I g.L-1 NaF solution （positive control）, deionized water （DDW, negative control）, and then 60-min application in pH 5.0 acidic buffer and 5-min application in neutral buffer. Acidic buffers were retained for calcium analysis. The main organic composition of GCE were GA and its isomer, and, to a lesser extent, small molecule gallotannins. The content of GA in GCE was 71.3%±0.2% （w/w）. Inorganic ions were present in various amounts, of which Ca was （136±2.82） μg.g-1, and Zn was （6.8±0.1） μg.g-1. No F was detected in GCE. in pH cycling, GA showed an effect similar to GCE in inhibiting enamel demineralization （P〉0.05）. GA was found to be the main effective, demineralization inhibiting component of GCE and could be a promising agent for the development of anticaries agents.

关键词： active component Galla chinensis gallic acid composition dental caries

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Oral health status in Sichuan Province: findings from the oral health survey of Sichuan, 2015–2016

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International Journal of Oral Science 2017年第1期9卷 10-15页

作者： Wei Yin Ying-Ming Yang Hong Chen Xue Li Zhuo Wang Li Cheng Qiu-Dan Yin Hong-Zhi Fang Wei Fei Fang-Lin Mi Min-Hai Nie Tao Hu Xue-Dong Zhou State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesDepartment of Preventive DentistryWest China Hospital of StomatologySichtJan UniversityChengduChina Sichuan Center for Disease Control and Prevention ChengduChina The Third People's Hospital of Chengdu ChengduChina Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital ChengduChina Department of Stomatolosy North Sichuan Medical CollegeFirst Affiliated HospitalNanchongChina Department of Stomatology Southwest Medical UniversityLuzhouChina and State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesWest China Hospital of StomatolosySichuan UniversityChengduChina

To investigate oral health status in the residents of Sichuan Province, southwest China, a cross-sectional study was performed using the latest Oral Health Survey Basic Methods recommended by the World Health Organization. A multistage stratified random cluster-sampling method was used to enroll participants from the following three groups： children aged 3-5 years, adolescents aged 12 years, and people aged 65-74 years. In these three groups, the mean numbers of teeth that were affected by caries were 3.28, 0.86 and 5.13, respectively, resulting in a prevalence of 63.47%, 37.20% and 83.20%, respectively. Relative to the high rate of decayed teeth, the prevalence of fillings was very low in all age groups （0.97%, 7.24% and 5.43%, respectively）, In the 12-year-old adolescent group, only 3.61% had good pit and fissure sealing. In addition, the rate of dental fluorosis was 24.80%, and the Community Fluorosis Index value was 0.39. In the elder group, the community periodontal index was 2.92. The prevalence in the elderly of having lost at least one tooth was 75.54%. Additionally, 4.44% of these participants were edentulous. The incidence of dental prosthesis was 51.75%, the proportion with a removable partial denture, a fixed denture, full dentures, dental implants and an informal fixed bridge was 21.59%, 11.45%, 4.64%, 0 and 16.67%, respectively. In this study, 8.2% of the elderly participants were affected by different types of oral mucosal lesions. Among such lesions, recurrent aphthous ulcers were most prevalent （2.69%） and oral lichen planuses were second （1.6%）. The conclusion presented in this survey is that oral diseases, especially dental caries and periodontal disease, are frequent and common in Sichuan province, China. Moreover, the treatment rate is very low, and primary prevention and treatment options are therefore urgently needed in this population.

关键词： cross-sectional study dental caries oral health status periodontal condition Sichuan Province of China

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Evaluation of three-dimensional biofilms on antibacterial bonding agents containing novel quaternary ammonium methacrylates

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International Journal of Oral Science 2014年第2期6卷 77-86页

作者： Han Zhou Michael D Weir Joseph M Antonucci Gary E Schumacher Xue-Dong Zhou Hockin H K Xu Department of Endodontics Prosthodontics and Operative Dentistry University of Maryland Dental School Baltimore USA State Key Laboratory of Oral Diseases West China Hospital of Stomatology Sichuan University Biomaterials Group Biosystems and Biomaterials Division National Institute of Standards and TechnologyGaithersburg USA Dr. Anthony Volpe Research Center American Dental Association Foundation Gaithersburg USA Center for Stem Cell Biology & Regenerative Medicine University of Maryland School of Medicine Baltimore USA Department of Mechanical Engineering University of Maryland Baltimore County USA

Antibacterial adhesives are promising to inhibit biofilms and secondary caries. The objectives of this study were to synthesize and incorporate quaternary ammonium methacrylates into adhesives, and investigate the alkyl chain length effects on three-dimensional biofilms adherent on adhesives for the first time. Six quaternary ammonium methacrylates with chain lengths of 3, 6, 9, 12, 16 and 18 were synthesized and incorporated into Scotchbond Multi-Purpose. Streptococcus mutans bacteria were cultured on resin to form biofilms. Confocal laser scanning microscopy was used to measure biofilm thickness, live/dead volumes and live-bacteria percentage vs. distance from resin surface. Biofilm thickness was the greatest for Scotchbond control; it decreased with increasing chain length, reaching a minimum at chain length 16. Live-biofilm volume had a similar trend. Dead-biofilm volume increased with increasing chain length. The adhesive with chain length 9 had 37% live bacteria near resin surface, but close to 100% live bacteria in the biofilm top section. For chain length 16, there were nearly 0% live bacteria throughout the three-dimensional biofilm. In conclusion, strong antibacterial activity was achieved by adding quaternary ammonium into adhesive, with biofilm thickness and live-biofilm volume decreasing as chain length was increased from 3 to 16. Antibacterial adhesives typically only inhibited bacteria close to its surface; however, adhesive with chain length 16 had mostly dead bacteria in the entire three-dimensional biofilm. Antibacterial adhesive with chain length 16 is promising to inhibit biofilms at the margins and combat secondary caries.

关键词： alkyl chain length antibacterial bondingagent dental caries quaternaryammonium methacrylate Streptococcusmutans three-dimensional biofilm

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Role of sortase in Streptococcus mutans under the effect of nicotine

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International Journal of Oral Science 2013年第4期5卷 206-211页

作者： Ming-Yun Li Rui-Jie Huang Xue-Dong Zhou Richard L Gregory School of Dentistry Indiana University State Key Laboratory of Oral Diseases West China Hospital of StomatologySichuan University Department of Pathology and Laboratory Medicine School of MedicineIndiana University

Streptococcus mutans is a common Gram-positive bacterium and plays a significant role in dental caries. Tobacco and/or nicotine have documented effects on S. mutans growth and colonization. Sortase A is used by many Gram-positive bacteria, including S. mutans, to facilitate the insertion of certain cell surface proteins, containing an LPXTGX motif such as antigen 1/11. This study examined the effect of nicotine on the function of sortase A to control the physiology and growth of S. mutans using wild-type S. mutans NG8, and its isogenic sortase-defective and -complemented strains. Briefly, the strains were treated with increasing amounts of nicotine in planktonic growth, biofilm metabolism, and sucrose-induced and saliva-induced antigen I/ll-dependent biofilm formation assays. The strains exhibited no significant differences with different concentrations of nicotine in planktonic growth assays. However, they had significantly increased （P~〈 0.05） biofilm metabolic activity （2- to 3-fold increase） as the concentration of nicotine increased. Furthermore, the sortase-defective strain was more sensitive metabolically to nicotine than the wild-type or sortase-complemented strains. All strains had significantly increased sucrose-induced biofilm formation （2- to 3-fold increase） as a result of increasing concentrations of nicotine. However, the sortase-defective strain was not able to make as much sucrose- and saliva-induced biofilm as the wild-type NG8 did with increasing nicotine concentrations. These results indicated that nicotine increased metabolic activity and sucrose-induced biofilm formation. The saliva-induced biofilm formation assay and qPCR data suggested that antigen 1/11 was upregulated with nicotine but biofilm was not able to be formed as much as wild-type NG8 without functional sortase A.

关键词： dental caries metabolism nicotine sortase A Streptococcus mutans

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Oral Microbiology:Past,Present and Future

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International Journal of Oral Science 2009年第2期1卷 47-58页

作者： Xue-song He Wen-yuan Shi School of Dentistry the University of California Los Angeles USA

Since the initial observations of oral bacteria within dental plaque by van Leeuwenhoek using his primitive microscopes in 1680, an event that is generally recognized as the advent of oral microbiological investigation, oral microbiology has gone through phases of ＂reductionism＂ and ＂holism＂. From the small beginnings of the Miller and Black period, in which microbiologists followed Koch＇s postulates, took the reductionist approach to try to study the complex oral microbial community by analyzing individual species; to the modem era when oral researchers embrace ＂holism＂ or ＂system thinking＂, adopt new concepts such as interspecies interaction, microbial community, biofilms, poly-microbial diseases, oral microbiological knowledge has burgeoned and our ability to identify the resident organisms in dental plaque and decipher the interactions between key components has rapidly increased, such knowledge has greatly changed our view of the oral microbial flora, provided invaluable insight into the etiology of dental and periodontal diseases, opened the door to new approaches and techniques for developing new therapeutic and preventive tools for combating oral polymicrobial diseases.

关键词： oral microbiology biofilms dental caries oral pathogenesis

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