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 Organiza...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 was2.92. 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.展开更多
Both bone morphogenetic protein 2(BMP2) and the wingless-type MMTV integration site(WNT)/p-catenin signalling pathway play important roles in odontoblast differentiation and dentinogenesis.Cross-talk between BMP2 and ...Both bone morphogenetic protein 2(BMP2) and the wingless-type MMTV integration site(WNT)/p-catenin signalling pathway play important roles in odontoblast differentiation and dentinogenesis.Cross-talk between BMP2 and WNT/p-catenin in osteoblast differentiation and bone formation has been identified.However,the roles and mechanisms of the canonical WNT pathway in the regulation of BMP2 in dental pulp injury and repair remain largely unknown.Here,we demonstrate that BMP2 promotes the differentiation of human dental pulp cells(HDPCs) by activating WNT/p-catenin signalling,which is further mediated by p38mitogen-activated protein kinase(MAPK) in vitro.BMP2 stimulation upregulated the expression of p-catenin in HDPCs,which was abolished by SB203580 but not by Noggin or LDN193189.Furthermore,BMP2 enhanced cell differentiation,which was not fully inhibited by Noggin or LDN193189.Instead,SB203580 partially blocked BMP2-induced p-catenin expression and cell differentiation.Taken together,these data suggest a possible mechanism by which the elevation of p-catenin resulting from BMP2 stimulation is mediated by the p38 MAPK pathway,which sheds light on the molecular mechanisms of BMP2-mediated pulp reparative dentin formation.展开更多
To determine the chemical composition of Galla chinensis extract(GCE) by several analysis techniques and to compare the efficacy of GCE and its main component(s) in inhibition of enamel demineralization,for the develo...To determine the chemical composition of Galla chinensis extract(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:4 g?L 21 GCE solution,4 g?L 21 gallic acid(GA) solution,1 g?L 21 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%60.2%(w/w).Inorganic ions were present in various amounts,of which Ca was(13662.82) mg?g 21,and Zn was(6.860.1) mg?g 21.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.展开更多
Dental composites are commonly used restorative materials; however, secondary caries due to bio?lm acids remains a major problem. The objectives of this study were(1) to develop a composite containing quaternary ammon...Dental composites are commonly used restorative materials; however, secondary caries due to bio?lm acids remains a major problem. The objectives of this study were(1) to develop a composite containing quaternary ammonium dimethacrylate(QADM),nanoparticles of silver(NAg), and nanoparticles of amorphous calcium phosphate(NACP), and(2) to conduct the ?rst investigation of the mechanical properties, bio?lm response and acid production vs water-ageing time from 1 day to 12 months.A 4 × 5 design was utilized, with four composites(NACP-QADM composite, NACP-NAg composite, NACP-QADM-NAg composite,and a commercial control composite), and ?ve water-ageing time periods(1 day, and 3, 6, 9, and 12 months). After each waterageing period, the mechanical properties of the resins were measured in a three-point ?exure, and antibacterial properties were tested via a dental plaque bio?lm model using human saliva as an inoculum. After 12 months of water-ageing, NACP-QADMNAg had a ?exural strength and elastic modulus matching those of the commercial control(P40.1). Incorporation of QADM or NAg into the NACP composite greatly reduced bio?lm viability, metabolic activity and acid production. A composite containing both QADM and NAg possessed a stronger antibacterial capability than one with QADM or NAg alone(Po0.05). The anti-bio?lm activity was maintained after 12 months of water-ageing and showed no signi?cant decrease with increasing time(P40.1). In conclusion, the NACP-QADM-NAg composite decreased bio?lm viability and lactic acid production, while matching the loadbearing capability of a commercial composite. There was no decrease in its antibacterial properties after 1 year of water-ageing.The durable antibacterial and mechanical properties indicate that NACP-QADM-NAg composites may be useful in dental restorations to combat caries.展开更多
Tooth bleaching agents may weaken the tooth structure. Therefore, it is important to minimize any risks of tooth hard tissue damage caused by bleaching agents. The aim of this study was to evaluate the effects of appl...Tooth bleaching agents may weaken the tooth structure. Therefore, it is important to minimize any risks of tooth hard tissue damage caused by bleaching agents. The aim of this study was to evaluate the effects of applying 45S5 bioglass (BG) before, after, and during 35% hydrogen peroxide (HP) bleaching on whitening efficacy, physicochemical properties and microstructures of bovine enamel. Seventy-two bovine enamel blocks were prepared and randomly divided into six groups: distilled deionized water (DDW), BG, HP, BG before HP, BG after HP and BG during HP. Colorimetric and microhardness tests were performed before and after the treatment procedure. Representative specimens from each group were selected for morphology investigation after the final tests. A significant color change was observed in group HP, BG before HP, BG after HP and BG during HP. The microhardness loss was in the following order: group HP>BG before HP, BG after HP>BG during HP>DDW, BG. The most obvious morphological alteration of was observed on enamel surfaces in group HP, and a slight morphological alteration was also detected in group BG before HP and BG after HP. Our findings suggest that the combination use of BG and HP could not impede the tooth whitening efficacy. Using BG during HP brought better protective effect than pre/post-bleaching use of BG, as it could more effectively reduce the mineral loss as well as retain the surface integrity of enamel. BG may serve as a promising biomimetic adjunct for bleaching therapy to prevent/restore the enamel damage induced by bleaching agents.展开更多
The aim of this study was to analyze the specific influence of root canal anatomy on the accessibility of working length during root canal therapy. Four hundred seventy-six root canal therapy cases (amounting to a tot...The aim of this study was to analyze the specific influence of root canal anatomy on the accessibility of working length during root canal therapy. Four hundred seventy-six root canal therapy cases (amounting to a total of 1 005 root canals) were examined. The anatomy risk factors assessed in each case included: tooth type (tooth location), root canal curvature, and root canal calcification, as well as endodontic retreatment. The investigation examined the correlation between each of these anatomic factors and the working length, with statistical analysis consisting of Chi-square tests and multiple logistic regression analysis. In an independent factor analysis, tooth type (tooth location), root canal curvature, canal calcification, and endodontic retreat- ment were determined to be the primary risk factors. In a multiple-factor regression model, root curvature and canal calcification were found to most significantly influence root canal working length accessibility (P<0.05). Root canal anatomy increases the difficulty of root canal preparation. Appropriate consideration of tooth anatomy will assist in accurate determination of preparation difficulty before instrumentation. This study alerts clinical therapists to anatomical factors influencing the working length accessibility, and allows for a direct estimate of success rate given in situ measurements of tooth factors during the root canal treatment procedure.展开更多
Osteoporosis is a serious public bone metabolic disease. However, the mechanisms underlying bone loss combined with ageing,which is known as senile osteoporosis, remains unknown. Here we show the detailed phenotype of...Osteoporosis is a serious public bone metabolic disease. However, the mechanisms underlying bone loss combined with ageing,which is known as senile osteoporosis, remains unknown. Here we show the detailed phenotype of this disease caused by SIRT6 knock out(KO) in mice. To the best of our knowledge, this is the first study to reveal that SIRT6 is expressed in both bone marrow stroma cells and bone-related cells in both mouse and human models, which suggests that SIRT6 is an important regulator in bone metabolism. SIRT6-KO mice exhibit a significant decrease in body weight and remarkable dwarfism. The skeleton of the SIRT6-KO mouse is deficient in cartilage and mineralized bone tissue. Moreover, the osteocalcin concentration in blood is lower, which suggests that bone mass is markedly lost. Besides, the tartrate-resistant acid phosphatase 5b(TRAP5b)concentration is much higher, which suggests that bone resorption is overactive. Both trabecular and cortical bones exhibit severe osteopenia, and the bone mineral density is decreased. Moreover, double-labelling analysis shows that bone formation is much slower. To determine whether SIRT6 directly regulates bone metabolism, we cultured primary bone marrow stromal cells for osteogenesis and osteoclastogenesis separately to avoid indirect interference in vivo responses such as inflammation. Taken together, these results show that SIRT6 can directly regulate osteoblast proliferation and differentiation, resulting in attenuation in mineralization. Furthermore, SIRT6 can directly regulate osteoclast differentiation and results in a higher number of small osteoclasts, which may be related to overactive bone resorption.展开更多
‘Bronze teeth’reflect the mechanical properties of natural teeth to a certain extent.Their mechanical properties resemble those of a tough metal,and the gradient of these properties lies in the direction from outsid...‘Bronze teeth’reflect the mechanical properties of natural teeth to a certain extent.Their mechanical properties resemble those of a tough metal,and the gradient of these properties lies in the direction from outside to inside.These attributes confer human teeth with effective mastication ability.Understanding the various mechanical properties of human teeth and dental materials is the basis for the development of restorative materials.In this study,the elastic properties,dynamic mechanical properties(visco-elasticity)and fracture mechanical properties of enamel and dentin were reviewed to provide a more thorough understanding of the mechanical properties of human teeth.展开更多
Saliva is secreted from the salivary glands and has multiple functions, including mouth cleaning and protection, antibacterial effects and digestion. With the rapid advancement in salivaomics, saliva is well recognize...Saliva is secreted from the salivary glands and has multiple functions, including mouth cleaning and protection, antibacterial effects and digestion. With the rapid advancement in salivaomics, saliva is well recognized as a pool of biological markers.Saliva, as a non-invasive and safe source, could be a substitute for blood in the diagnosis and prognosis of diseases. This review summarizes the latest advancements in saliva-related studies and addresses the potential value of saliva in the early diagnosis of oral diseases, such as dental caries and periodontal disease, as well as cancer, diabetes and other systemic disorders. Saliva biomarkers range from changes in the biochemical indices of DNA, RNA and proteins to the diversi?cation of microbiota structures. This study integrates data reported in the recent literature and discusses the clinical signi?cance and prospects for the application of saliva in the early diagnosis of diseases, translational medicine and precision medicine.展开更多
Dental restorative materials with antimicrobial properties can inhibit bacterial colonization, which may result in a reduction of caries at tooth-filling interaction zones. This study aimed to develop antibacterial gl...Dental restorative materials with antimicrobial properties can inhibit bacterial colonization, which may result in a reduction of caries at tooth-filling interaction zones. This study aimed to develop antibacterial glass–ionomer cements(GIC) containing a quaternary ammonium monomer(dimethylaminododecyl methacrylate, DMADDM), and to investigate their effect on material performance and antibacterial properties. Different mass fractions(0, 1.1% and 2.2%) of DMADDM were incorporated into the GIC. The flexure strength, surface charge density, surface roughness and fluoride release were tested. A Streptococcus mutans biofilm model was used. Exopolysaccharides(EPS) staining was used to analyze the inhibitory effect of DMADDM on the biofilm matrix. In addition, biofilm metabolic activity, lactic acid metabolism and the expression of glucosyltransferase genes gtf B, gtf C and gtf D were measured. GIC containing 1.1% and 2.2% DMADDM had flexural strengths matching those of the commercial control(P40.1). DMADDM was able to increase the surface charge density but reduced surface roughness(Po0.05). The incorporation of 1.1% and 2.2% DMADDM elevated the release of fluoride by the GIC in the first 2 days(Po0.05). The novel DMADDM-modified GIC significantly reduced biofilm metabolic activity(Po0.05) and decreased lactic acid production(Po0.05). The quantitative polymerase chain reaction(q PCR) results showed that the expression of gtf B, gtf C and gtf D decreased when mass fractions of DMADDM increased(Po0.05). EPS staining showed that both the bacteria and EPS in biofilm decreased in the DMADDM groups. The incorporation of DMADDM could modify the properties of GIC to influence the development of S. mutans biofilms. In this study, we investigated the interface properties of antibacterial materials for the first time. GIC containing DMADDM can improve material performance and antibacterial properties and may contribute to the better management of secondary caries.展开更多
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...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 18were 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.展开更多
Antibacterial dimethylaminododecyl methacrylate(DMADDM) was recently synthesized. The objectives of this study were to:(1)investigate antibacterial activity of DMADDM-containing primer on Streptococcus mutans impregna...Antibacterial dimethylaminododecyl methacrylate(DMADDM) was recently synthesized. The objectives of this study were to:(1)investigate antibacterial activity of DMADDM-containing primer on Streptococcus mutans impregnated into dentin blocks for the first time, and(2) compare the antibacterial efficacy of DMADDM with a previous quaternary ammonium dimethacrylate(QADM).Scotchbond Multi-Purpose(SBMP) bonding agent was used. DMADDM and QADM were mixed into SBMP primer. Six primers were tested: SBMP control primer P, P+2.5% DMADDM, P+5% DMADDM, P+7.5% DMADDM, P+10% DMADDM, and P+10% QADM.S. mutans were impregnated into human dentin blocks, and each primer was applied to dentin to test its ability to kill bacteria in dentinal tubules. Bacteria in dentin were collected via a sonication method, and the colony-forming units(CFU) and inhibition zones were measured. The bacterial inhibition zone of P+10% DMADDM was 10 times that of control primer(Po0.05). CFU in dentin with P+10% DMADDM was reduced by three orders of magnitude, compared with control. DMADDM had a much stronger antibacterial effect than QADM, and antibacterial efficacy increased with increasing DMADDM concentration. Dentin shear bond strengths were similar among all groups(P40.1). In conclusion, antibacterial DMADDM-containing primer was validated to kill bacteria inside dentin blocks, possessing a much stronger antibacterial potency than the previous QADM. DMADDM-containing bonding agent was effective in eradicating bacteria in dentin, and its efficacy was directly proportional to DMADDM mass fraction. Therefore, DMADDM may be promising for use in bonding agents as well as in other restorative and preventive materials to inhibit bacteria.展开更多
Marginal bone loss during bone healing exists around non-submerged dental implants. The aim of this study was to identify the relationship between different degrees of marginal bone loss during bone healing and the sa...Marginal bone loss during bone healing exists around non-submerged dental implants. The aim of this study was to identify the relationship between different degrees of marginal bone loss during bone healing and the salivary microbiome. One hundred patients were recruited, and marginal bone loss around their implants was measured using cone beam computed tomography during a 3-month healing period. The patients were divided into three groups according to the severity of marginal bone loss.Saliva samples were collected from all subjected and were analysed using 16 SMiSeq sequencing. Although the overall structure of the microbial community was not dramatically altered, the relative abundance of several taxonomic groups noticeably changed. The abundance of species in the phyla Spirochaeta and Synergistetes increased significantly as the bone loss became more severe. Species within the genus Treponema also exhibited increased abundance, whereas Veillonella, Haemophilus and Leptotrichia exhibited reduced abundances, in groups with more bone loss. Porphyromonasgingivalis, Treponemadenticola and Streptococcus intermedius were significantly more abundant in the moderate group and/or severe group. The severity of marginal bone loss around the non-submerged implant was associated with dissimilar taxonomic compositions. An increased severity of marginal bone loss was related to increased proportions of periodontal pathogenic species. These data suggest a potential role of microbes in the progression of marginal bone loss during bone healing.展开更多
Salivary analysis can be used to assess the severity of caries. Of the known salivary proteins, a paucity of information exists concerning the role of proteinase 3(PR3), a serine protease of the chymotrypsin family, i...Salivary analysis can be used to assess the severity of caries. Of the known salivary proteins, a paucity of information exists concerning the role of proteinase 3(PR3), a serine protease of the chymotrypsin family, in dental caries. Whole, unstimulated saliva was collected from children with varying degrees of active caries and tested using a Human Protease Array Kit and an enzyme-linked immunosorbent assay.A significantly decreased concentration of salivary PR3 was noted with increasing severity of dental caries(P,0.01); a positive correlation(r50.87; P,0.01; Pearson's correlation analysis) was also observed between salivary p H and PR3 concentration. In an antibacterial test,a PR3 concentration of 250 ng?m L21 or higher significantly inhibited Streptococcus mutans UA159 growth after 12 h of incubation(P,0.05). These studies indicate that PR3 is a salivary factor associated with the severity of dental caries, as suggested by the negative relationship between salivary PR3 concentration and the severity of caries as well as the susceptibility of S. mutans to PR3.展开更多
D-alanine(D-Ala) is an essential amino acid that has a key role in bacterial cell wall synthesis. Alanine racemase(Alr) is a unique enzyme that interconverts L-alanine and D-alanine in most bacteria, making this enzym...D-alanine(D-Ala) is an essential amino acid that has a key role in bacterial cell wall synthesis. Alanine racemase(Alr) is a unique enzyme that interconverts L-alanine and D-alanine in most bacteria, making this enzyme a potential target for antimicrobial drug development. Streptococcus mutans is a major causative factor of dental caries. The factors involved in the survival, virulence and interspecies interactions of S. mutans could be exploited as potential targets for caries control. The current study aimed to investigate the physiological role of Alr in S. mutans. We constructed alr mutant strain of S. mutans and evaluated its phenotypic traits and interspecies competitiveness compared with the wild-type strain. We found that alr deletion was lethal to S. mutans. A minimal supplement of D-Ala(150 μg·m L^(-1)) was required for the optimal growth of the alr mutant.The depletion of D-alanine in the growth medium resulted in cell wall perforation and cell lysis in the alr mutant strain. We also determined the compromised competitiveness of the alr mutant strain relative to the wild-type S. mutans against other oral streptococci(S. sanguinis or S. gordonii), demonstrated using either conditioned medium assays or dual-species fluorescent in situ hybridization analysis. Given the importance and necessity of alr to the growth and competitiveness of S. mutans, Alr may represent a promising target to modulate the cariogenicity of oral biofilms and to benefit the management of dental caries.展开更多
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 G...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 I/II. 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/II-dependent biofilm formation assays. The strains exhibited no significant differences with different concentrations of nicotine in planktonic growth assays. However, they had significantly increased(Pf0.05) 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 I/II was upregulated with nicotine but biofilm was not able to be formed as much as wild-type NG8 without functional sortase A.展开更多
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 ...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 preve Fnt 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 μg·m L^(-1), 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 m M, 1 m M and 10 m M of extra glucose significantly reduced the number of persisters both in planktonic and biofilm conditions. The formation of noninheritable and multidrug tolerant persisters induced by DMADDM suggested that drug tolerance and new persistent eradication strategies should be considered for oral antibacterial agents.展开更多
Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. ...Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. To explore the spatiotemporal expression of histone H3 lysine 4trimethylation(H3K4me3) and histone H3 lysine 27 trimethylation(H3K27me3) epigenetic marks and methylation or demethylation transferases in tooth organ development, we measured the expression of SET7, EZH2, KDM5 B and JMJD3 via immunohistochemistry and quantitative polymerase chain reaction(qP CR) analysis in the first molar of BALB/c mice embryos at E13.5, E15.5, E17.5, P0 and P3, respectively. We also measured the expression of H3K4me3 and H3K27me3 with immunofluorescence staining. During murine tooth germ development, methylation or demethylation transferases were expressed in a spatial–temporal manner. The bivalent modification characterized by H3K4me3 and H3K27me3 can be found during the tooth germ development, as shown by immunofluorescence. The expression of SET7, EZH2 as methylation transferases and KDM5 B and JMJD3 as demethylation transferases indicated accordingly with the expression of H3K4me3 and H3K27me3 respectively to some extent. The bivalent histone may play a critical role in tooth organ development via the regulation of cell differentiation.展开更多
Ever since the first discovery of bacteria over 400 years ago by van LeeuwenhoeK,reductionism has been used by microbiologists as they analyzed small components of individually isolated bacteria to try and understand ...Ever since the first discovery of bacteria over 400 years ago by van LeeuwenhoeK,reductionism has been used by microbiologists as they analyzed small components of individually isolated bacteria to try and understand the whole.Powered by molecular biology and genomics,modern microbiologists have realized that the whole is more than the simple sum of its parts."System thinking"and"holism"have led microbiologists from studying individual cells to examining complex communities.Biofilm research is at the center stage of this exciting new revolution!展开更多
Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise co...Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol·L-1 glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.展开更多
基金funded by a grant from Scientific Research in the Public Interest(201502002)conducted by the Department of Preventive Dentistry,West China Hospital of Stomatology,Sichuan University,China
文摘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 was2.92. 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.
基金supported by the National Nature Science Foundation of China(grant nos.81200759,81070801 and 813220170)the Innovative Research Team of the Education Department of Sichuan Province(13TD0038)+1 种基金the Sichuan Province Science and Technology Support Program(2012SZ0034)the Program of International Science and Technology Cooperation(2014DFA31990)
文摘Both bone morphogenetic protein 2(BMP2) and the wingless-type MMTV integration site(WNT)/p-catenin signalling pathway play important roles in odontoblast differentiation and dentinogenesis.Cross-talk between BMP2 and WNT/p-catenin in osteoblast differentiation and bone formation has been identified.However,the roles and mechanisms of the canonical WNT pathway in the regulation of BMP2 in dental pulp injury and repair remain largely unknown.Here,we demonstrate that BMP2 promotes the differentiation of human dental pulp cells(HDPCs) by activating WNT/p-catenin signalling,which is further mediated by p38mitogen-activated protein kinase(MAPK) in vitro.BMP2 stimulation upregulated the expression of p-catenin in HDPCs,which was abolished by SB203580 but not by Noggin or LDN193189.Furthermore,BMP2 enhanced cell differentiation,which was not fully inhibited by Noggin or LDN193189.Instead,SB203580 partially blocked BMP2-induced p-catenin expression and cell differentiation.Taken together,these data suggest a possible mechanism by which the elevation of p-catenin resulting from BMP2 stimulation is mediated by the p38 MAPK pathway,which sheds light on the molecular mechanisms of BMP2-mediated pulp reparative dentin formation.
基金supported by the National Natural Science Foundation of China (grant nos. 30572409 and 30430800)Youth Foundation of Sichuan Province in China (grant no. 07ZQ026-020)
文摘To determine the chemical composition of Galla chinensis extract(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:4 g?L 21 GCE solution,4 g?L 21 gallic acid(GA) solution,1 g?L 21 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%60.2%(w/w).Inorganic ions were present in various amounts,of which Ca was(13662.82) mg?g 21,and Zn was(6.860.1) mg?g 21.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.
基金supported by the International Science and Technology Cooperation Program of China (2014DFE30180) (Xue-Dong Zhou)National Natural Science Foundation of China grant 81430011 (Xue-Dong Zhou),81372889 (Lei Cheng),81400540 (Ke Zhang)+1 种基金?nancial support from the School of Stomatology at the Capital Medical University in China (Ke Zhang),NIH R01 DE17974 (Hockin HK Xu)a seed grant from the University of Maryland School of Dentistry (Hockin HK Xu)
文摘Dental composites are commonly used restorative materials; however, secondary caries due to bio?lm acids remains a major problem. The objectives of this study were(1) to develop a composite containing quaternary ammonium dimethacrylate(QADM),nanoparticles of silver(NAg), and nanoparticles of amorphous calcium phosphate(NACP), and(2) to conduct the ?rst investigation of the mechanical properties, bio?lm response and acid production vs water-ageing time from 1 day to 12 months.A 4 × 5 design was utilized, with four composites(NACP-QADM composite, NACP-NAg composite, NACP-QADM-NAg composite,and a commercial control composite), and ?ve water-ageing time periods(1 day, and 3, 6, 9, and 12 months). After each waterageing period, the mechanical properties of the resins were measured in a three-point ?exure, and antibacterial properties were tested via a dental plaque bio?lm model using human saliva as an inoculum. After 12 months of water-ageing, NACP-QADMNAg had a ?exural strength and elastic modulus matching those of the commercial control(P40.1). Incorporation of QADM or NAg into the NACP composite greatly reduced bio?lm viability, metabolic activity and acid production. A composite containing both QADM and NAg possessed a stronger antibacterial capability than one with QADM or NAg alone(Po0.05). The anti-bio?lm activity was maintained after 12 months of water-ageing and showed no signi?cant decrease with increasing time(P40.1). In conclusion, the NACP-QADM-NAg composite decreased bio?lm viability and lactic acid production, while matching the loadbearing capability of a commercial composite. There was no decrease in its antibacterial properties after 1 year of water-ageing.The durable antibacterial and mechanical properties indicate that NACP-QADM-NAg composites may be useful in dental restorations to combat caries.
基金supported by the Research Fund from Science and Technology Department of Sichuan Province (No. 2009FZ0065)Key Project of the Science and Technology Department of Sichuan Province (No. 2011SZ0101)+1 种基金Doctoral Fund of Ministry of Education of China (No. 20120181120002)supported by Open Fund of State Key Laboratory of Oral Diseases, Sichuan University
文摘Tooth bleaching agents may weaken the tooth structure. Therefore, it is important to minimize any risks of tooth hard tissue damage caused by bleaching agents. The aim of this study was to evaluate the effects of applying 45S5 bioglass (BG) before, after, and during 35% hydrogen peroxide (HP) bleaching on whitening efficacy, physicochemical properties and microstructures of bovine enamel. Seventy-two bovine enamel blocks were prepared and randomly divided into six groups: distilled deionized water (DDW), BG, HP, BG before HP, BG after HP and BG during HP. Colorimetric and microhardness tests were performed before and after the treatment procedure. Representative specimens from each group were selected for morphology investigation after the final tests. A significant color change was observed in group HP, BG before HP, BG after HP and BG during HP. The microhardness loss was in the following order: group HP>BG before HP, BG after HP>BG during HP>DDW, BG. The most obvious morphological alteration of was observed on enamel surfaces in group HP, and a slight morphological alteration was also detected in group BG before HP and BG after HP. Our findings suggest that the combination use of BG and HP could not impede the tooth whitening efficacy. Using BG during HP brought better protective effect than pre/post-bleaching use of BG, as it could more effectively reduce the mineral loss as well as retain the surface integrity of enamel. BG may serve as a promising biomimetic adjunct for bleaching therapy to prevent/restore the enamel damage induced by bleaching agents.
基金supported by State Key Laboratory of Oral Diseases and Department of Endodontics and Operative Dentistry, West China School of Stomatology, Sichuan Universitysupported by the Key Clinical Program of the Ministry of Health of China (2010)National Key Clinical Program of China (2010)
文摘The aim of this study was to analyze the specific influence of root canal anatomy on the accessibility of working length during root canal therapy. Four hundred seventy-six root canal therapy cases (amounting to a total of 1 005 root canals) were examined. The anatomy risk factors assessed in each case included: tooth type (tooth location), root canal curvature, and root canal calcification, as well as endodontic retreatment. The investigation examined the correlation between each of these anatomic factors and the working length, with statistical analysis consisting of Chi-square tests and multiple logistic regression analysis. In an independent factor analysis, tooth type (tooth location), root canal curvature, canal calcification, and endodontic retreat- ment were determined to be the primary risk factors. In a multiple-factor regression model, root curvature and canal calcification were found to most significantly influence root canal working length accessibility (P<0.05). Root canal anatomy increases the difficulty of root canal preparation. Appropriate consideration of tooth anatomy will assist in accurate determination of preparation difficulty before instrumentation. This study alerts clinical therapists to anatomical factors influencing the working length accessibility, and allows for a direct estimate of success rate given in situ measurements of tooth factors during the root canal treatment procedure.
基金supported by NSFC grants 81371136 and JCPT2011-9 (Xue-Dong Zhou),NSFC grants 81470711 and 81200760 (Li-Wei Zheng)Ling Ye (Professor, Sate Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University) for financial aid
文摘Osteoporosis is a serious public bone metabolic disease. However, the mechanisms underlying bone loss combined with ageing,which is known as senile osteoporosis, remains unknown. Here we show the detailed phenotype of this disease caused by SIRT6 knock out(KO) in mice. To the best of our knowledge, this is the first study to reveal that SIRT6 is expressed in both bone marrow stroma cells and bone-related cells in both mouse and human models, which suggests that SIRT6 is an important regulator in bone metabolism. SIRT6-KO mice exhibit a significant decrease in body weight and remarkable dwarfism. The skeleton of the SIRT6-KO mouse is deficient in cartilage and mineralized bone tissue. Moreover, the osteocalcin concentration in blood is lower, which suggests that bone mass is markedly lost. Besides, the tartrate-resistant acid phosphatase 5b(TRAP5b)concentration is much higher, which suggests that bone resorption is overactive. Both trabecular and cortical bones exhibit severe osteopenia, and the bone mineral density is decreased. Moreover, double-labelling analysis shows that bone formation is much slower. To determine whether SIRT6 directly regulates bone metabolism, we cultured primary bone marrow stromal cells for osteogenesis and osteoclastogenesis separately to avoid indirect interference in vivo responses such as inflammation. Taken together, these results show that SIRT6 can directly regulate osteoblast proliferation and differentiation, resulting in attenuation in mineralization. Furthermore, SIRT6 can directly regulate osteoclast differentiation and results in a higher number of small osteoclasts, which may be related to overactive bone resorption.
基金support from the National Natural Science Foundation of China(Nos.81170996,81070867 and 81100777)the Sichuan Province Science and Technology Innovation Team Program(2011JTD0006)
文摘‘Bronze teeth’reflect the mechanical properties of natural teeth to a certain extent.Their mechanical properties resemble those of a tough metal,and the gradient of these properties lies in the direction from outside to inside.These attributes confer human teeth with effective mastication ability.Understanding the various mechanical properties of human teeth and dental materials is the basis for the development of restorative materials.In this study,the elastic properties,dynamic mechanical properties(visco-elasticity)and fracture mechanical properties of enamel and dentin were reviewed to provide a more thorough understanding of the mechanical properties of human teeth.
基金supported by the National Natural Science Foundation of China (81430011 to Xue-Dong Zhou and 81371135 to Ji-Yao Li)Clinical Advanced Technique Award of West China Hospital of Stomatology (LCXJS2015 to Xin Xu)the Brilliant Young Investigator Award,Sichuan University (2015SCU04A16 to Xin Xu)
文摘Saliva is secreted from the salivary glands and has multiple functions, including mouth cleaning and protection, antibacterial effects and digestion. With the rapid advancement in salivaomics, saliva is well recognized as a pool of biological markers.Saliva, as a non-invasive and safe source, could be a substitute for blood in the diagnosis and prognosis of diseases. This review summarizes the latest advancements in saliva-related studies and addresses the potential value of saliva in the early diagnosis of oral diseases, such as dental caries and periodontal disease, as well as cancer, diabetes and other systemic disorders. Saliva biomarkers range from changes in the biochemical indices of DNA, RNA and proteins to the diversi?cation of microbiota structures. This study integrates data reported in the recent literature and discusses the clinical signi?cance and prospects for the application of saliva in the early diagnosis of diseases, translational medicine and precision medicine.
基金supported by a National Natural Science Foundation of China grant 81372889 (Lei Cheng), 81430011 (Xue-dong Zhou)the Program for New Century Excellent Talents in University (Lei Cheng)+3 种基金a Youth Grant of the Science and Technology Department of Sichuan Province, China 2014JQ0033 (Lei Cheng)the International Science and Technology Cooperation Program of China 2014DFE30180 (Xue-dong Zhou)NIH R01 DE17974 (Hockin HK Xu)a Seed Grant (Hockin HK Xu) from the University of Maryland School of Dentistry
文摘Dental restorative materials with antimicrobial properties can inhibit bacterial colonization, which may result in a reduction of caries at tooth-filling interaction zones. This study aimed to develop antibacterial glass–ionomer cements(GIC) containing a quaternary ammonium monomer(dimethylaminododecyl methacrylate, DMADDM), and to investigate their effect on material performance and antibacterial properties. Different mass fractions(0, 1.1% and 2.2%) of DMADDM were incorporated into the GIC. The flexure strength, surface charge density, surface roughness and fluoride release were tested. A Streptococcus mutans biofilm model was used. Exopolysaccharides(EPS) staining was used to analyze the inhibitory effect of DMADDM on the biofilm matrix. In addition, biofilm metabolic activity, lactic acid metabolism and the expression of glucosyltransferase genes gtf B, gtf C and gtf D were measured. GIC containing 1.1% and 2.2% DMADDM had flexural strengths matching those of the commercial control(P40.1). DMADDM was able to increase the surface charge density but reduced surface roughness(Po0.05). The incorporation of 1.1% and 2.2% DMADDM elevated the release of fluoride by the GIC in the first 2 days(Po0.05). The novel DMADDM-modified GIC significantly reduced biofilm metabolic activity(Po0.05) and decreased lactic acid production(Po0.05). The quantitative polymerase chain reaction(q PCR) results showed that the expression of gtf B, gtf C and gtf D decreased when mass fractions of DMADDM increased(Po0.05). EPS staining showed that both the bacteria and EPS in biofilm decreased in the DMADDM groups. The incorporation of DMADDM could modify the properties of GIC to influence the development of S. mutans biofilms. In this study, we investigated the interface properties of antibacterial materials for the first time. GIC containing DMADDM can improve material performance and antibacterial properties and may contribute to the better management of secondary caries.
基金supported by NIH R01 DE17974West China School of Stomatologya Seed Grant from Department of Endodontics,Prosthodontics and Operative Dentistry,University of Maryland
文摘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 18were 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.
基金supported by NIH R01 DE17974 (Hockin HK Xu)a scholarship from West China School of Stomatology (Chen Chen)+3 种基金Program for New Century Excellent Talents in University (Lei Cheng)Youth Grant of Science and Technology Department of Sichuan Province China (2014JQ0033) (Lei Cheng)a bridge fund from University of Maryland Baltimore School of Dentistry (Hockin HK Xu)a seed grant from University of Maryland Baltimore (Hockin HK Xu)
文摘Antibacterial dimethylaminododecyl methacrylate(DMADDM) was recently synthesized. The objectives of this study were to:(1)investigate antibacterial activity of DMADDM-containing primer on Streptococcus mutans impregnated into dentin blocks for the first time, and(2) compare the antibacterial efficacy of DMADDM with a previous quaternary ammonium dimethacrylate(QADM).Scotchbond Multi-Purpose(SBMP) bonding agent was used. DMADDM and QADM were mixed into SBMP primer. Six primers were tested: SBMP control primer P, P+2.5% DMADDM, P+5% DMADDM, P+7.5% DMADDM, P+10% DMADDM, and P+10% QADM.S. mutans were impregnated into human dentin blocks, and each primer was applied to dentin to test its ability to kill bacteria in dentinal tubules. Bacteria in dentin were collected via a sonication method, and the colony-forming units(CFU) and inhibition zones were measured. The bacterial inhibition zone of P+10% DMADDM was 10 times that of control primer(Po0.05). CFU in dentin with P+10% DMADDM was reduced by three orders of magnitude, compared with control. DMADDM had a much stronger antibacterial effect than QADM, and antibacterial efficacy increased with increasing DMADDM concentration. Dentin shear bond strengths were similar among all groups(P40.1). In conclusion, antibacterial DMADDM-containing primer was validated to kill bacteria inside dentin blocks, possessing a much stronger antibacterial potency than the previous QADM. DMADDM-containing bonding agent was effective in eradicating bacteria in dentin, and its efficacy was directly proportional to DMADDM mass fraction. Therefore, DMADDM may be promising for use in bonding agents as well as in other restorative and preventive materials to inhibit bacteria.
基金supported by grants from the National Natural Science Foundation of China(NSFC8137117381571001)+2 种基金State Key Laboratory of Oral Diseases(SKLOD201704)International Team for Implantology(Grant No.975_2014,Basel,Switzerland)to Quan Yuanthe National Key R&D Program of China during the 13th Five-Year Plan(2016YFC1102700)to Xue-Dong Zhou
文摘Marginal bone loss during bone healing exists around non-submerged dental implants. The aim of this study was to identify the relationship between different degrees of marginal bone loss during bone healing and the salivary microbiome. One hundred patients were recruited, and marginal bone loss around their implants was measured using cone beam computed tomography during a 3-month healing period. The patients were divided into three groups according to the severity of marginal bone loss.Saliva samples were collected from all subjected and were analysed using 16 SMiSeq sequencing. Although the overall structure of the microbial community was not dramatically altered, the relative abundance of several taxonomic groups noticeably changed. The abundance of species in the phyla Spirochaeta and Synergistetes increased significantly as the bone loss became more severe. Species within the genus Treponema also exhibited increased abundance, whereas Veillonella, Haemophilus and Leptotrichia exhibited reduced abundances, in groups with more bone loss. Porphyromonasgingivalis, Treponemadenticola and Streptococcus intermedius were significantly more abundant in the moderate group and/or severe group. The severity of marginal bone loss around the non-submerged implant was associated with dissimilar taxonomic compositions. An increased severity of marginal bone loss was related to increased proportions of periodontal pathogenic species. These data suggest a potential role of microbes in the progression of marginal bone loss during bone healing.
基金the support of the National Natural Science Foundation of China (No. 81372892)Sichuan Province Science and Technology Innovation Team Program (JCPT 2011-9)
文摘Salivary analysis can be used to assess the severity of caries. Of the known salivary proteins, a paucity of information exists concerning the role of proteinase 3(PR3), a serine protease of the chymotrypsin family, in dental caries. Whole, unstimulated saliva was collected from children with varying degrees of active caries and tested using a Human Protease Array Kit and an enzyme-linked immunosorbent assay.A significantly decreased concentration of salivary PR3 was noted with increasing severity of dental caries(P,0.01); a positive correlation(r50.87; P,0.01; Pearson's correlation analysis) was also observed between salivary p H and PR3 concentration. In an antibacterial test,a PR3 concentration of 250 ng?m L21 or higher significantly inhibited Streptococcus mutans UA159 growth after 12 h of incubation(P,0.05). These studies indicate that PR3 is a salivary factor associated with the severity of dental caries, as suggested by the negative relationship between salivary PR3 concentration and the severity of caries as well as the susceptibility of S. mutans to PR3.
基金financially supported through grants from the National Natural Science Foundation of China (81400501 to Ming-Yun Li, 81371135 to Ji-Yao Li and 81430011 to Xue-Dong Zhou)the International Science and Technology Cooperation Programme of China (2014DFE30180 to Xue-Dong Zhou)+1 种基金the Talented Young Investigator Award of Sichuan University (2082604184224 to Xin Xu)the Special Fund of State Key Laboratory of Oral Diseases, Sichuan University (SKLOD201525 to Ming-Yun Li)
文摘D-alanine(D-Ala) is an essential amino acid that has a key role in bacterial cell wall synthesis. Alanine racemase(Alr) is a unique enzyme that interconverts L-alanine and D-alanine in most bacteria, making this enzyme a potential target for antimicrobial drug development. Streptococcus mutans is a major causative factor of dental caries. The factors involved in the survival, virulence and interspecies interactions of S. mutans could be exploited as potential targets for caries control. The current study aimed to investigate the physiological role of Alr in S. mutans. We constructed alr mutant strain of S. mutans and evaluated its phenotypic traits and interspecies competitiveness compared with the wild-type strain. We found that alr deletion was lethal to S. mutans. A minimal supplement of D-Ala(150 μg·m L^(-1)) was required for the optimal growth of the alr mutant.The depletion of D-alanine in the growth medium resulted in cell wall perforation and cell lysis in the alr mutant strain. We also determined the compromised competitiveness of the alr mutant strain relative to the wild-type S. mutans against other oral streptococci(S. sanguinis or S. gordonii), demonstrated using either conditioned medium assays or dual-species fluorescent in situ hybridization analysis. Given the importance and necessity of alr to the growth and competitiveness of S. mutans, Alr may represent a promising target to modulate the cariogenicity of oral biofilms and to benefit the management of dental caries.
文摘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 I/II. 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/II-dependent biofilm formation assays. The strains exhibited no significant differences with different concentrations of nicotine in planktonic growth assays. However, they had significantly increased(Pf0.05) 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 I/II was upregulated with nicotine but biofilm was not able to be formed as much as wild-type NG8 without functional sortase A.
基金supported by the National Key Research and Development Program of China 2016YFC1102700(X.Z.)the National Natural Science Foundation of China grants 81372889(LC)+2 种基金81600858(BR)and 81430011(XZ)the Recruitment Program for Young Professionals(MF)the Youth Grant of the Science and Technology Department of Sichuan Province,China2017JQ0028(L.C.)
文摘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 preve Fnt 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 μg·m L^(-1), 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 m M, 1 m M and 10 m M of extra glucose significantly reduced the number of persisters both in planktonic and biofilm conditions. The formation of noninheritable and multidrug tolerant persisters induced by DMADDM suggested that drug tolerance and new persistent eradication strategies should be considered for oral antibacterial agents.
基金supported by National Science Foundation of China (Grant No. 81371136) to Xue-Dong ZhouNational Science Foundation of China (Grant No. 81200760 and 81470711) to Li-Wei Zheng
文摘Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. To explore the spatiotemporal expression of histone H3 lysine 4trimethylation(H3K4me3) and histone H3 lysine 27 trimethylation(H3K27me3) epigenetic marks and methylation or demethylation transferases in tooth organ development, we measured the expression of SET7, EZH2, KDM5 B and JMJD3 via immunohistochemistry and quantitative polymerase chain reaction(qP CR) analysis in the first molar of BALB/c mice embryos at E13.5, E15.5, E17.5, P0 and P3, respectively. We also measured the expression of H3K4me3 and H3K27me3 with immunofluorescence staining. During murine tooth germ development, methylation or demethylation transferases were expressed in a spatial–temporal manner. The bivalent modification characterized by H3K4me3 and H3K27me3 can be found during the tooth germ development, as shown by immunofluorescence. The expression of SET7, EZH2 as methylation transferases and KDM5 B and JMJD3 as demethylation transferases indicated accordingly with the expression of H3K4me3 and H3K27me3 respectively to some extent. The bivalent histone may play a critical role in tooth organ development via the regulation of cell differentiation.
文摘Ever since the first discovery of bacteria over 400 years ago by van LeeuwenhoeK,reductionism has been used by microbiologists as they analyzed small components of individually isolated bacteria to try and understand the whole.Powered by molecular biology and genomics,modern microbiologists have realized that the whole is more than the simple sum of its parts."System thinking"and"holism"have led microbiologists from studying individual cells to examining complex communities.Biofilm research is at the center stage of this exciting new revolution!
基金supported by Chinese State Scholarship Fund to R. WangUS National Institutes of Health (NIH) Grants DE020102 and GM95373 to W. Shi
文摘Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol·L-1 glycine. Genomic DNA was extracted, subjected to 16S rRNA PCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F. nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species.
