纳米银对牙本质表面粪肠球菌生物膜的杀菌作用

目的 研究纳米银对牙本质表面粪肠球菌生物膜的杀菌作用,为探寻新型有效的根管消毒药物提供实验依据.方法 制备牙本质片309个,随机分成3组,在牙本质表面形成粪肠球菌生物膜后,分别用0.1%纳米银溶液、1.313%次氯酸钠溶液和0.9%氯化钠溶液处理.3组分别于处理0(即处理前)、1、6、12、24 h时取出样本,应用活菌...     

纳米银对多菌种生物膜中粪肠球菌的杀菌作用

目的:研究纳米银对多菌种生物膜中粪肠球菌的杀菌作用.方法:用粪肠球菌、产黑色素普雷沃菌以及具核梭杆菌构建多菌种生物膜85 个.分别用0.1%(12~15 nm、100 nm)纳米银悬浊液以及2%的次氯酸钠溶液处理,然后通过qPCR方法检测样本中粪肠球菌的活菌量和粪肠球菌的细菌总量.配对t检验比较各组细菌样本临界循环数(Ct).结果:各个实验组使用和不使用PMA之间粪肠球菌的细菌计数有着差别(P=0.000);将3 个实验组使用和不使用PMA qPCR检测到的Ct值的差值进行比较:纳米银(12~15 nm)实验组最大,纳米银(100 nm)实验组次之,次氯酸钠实验组最小(P<0.05).结论:纳米银对于多菌种生物膜中的粪肠球菌有着较强的杀菌作用,直径较小的纳米银杀菌作用较强.     

电解质水对粪肠球菌生物膜的杀菌作用研究

目的:观察强酸性电解质水(SAEW)和碱性电解质水(AEW)对粪肠球菌生物膜的杀菌作用。方法:利用BioFluxTM200微流体系统建立粪肠球菌生物膜模型,并分别用SAEW、AEW、52.5 g/L次氯酸钠液(阳性对照)和生理盐水(阴性对照)处理1、5、10 min后,激光共聚焦显微镜(CLSM)观察并采集荧光图像;ImageJ图像分析软件分析并计算各处理组不同时间点的活菌比例。结果:SAEW、AEW和52.5 g/L次氯酸钠液处理1、5、10 min各时间点的活菌比例都显著低于生理盐水组(P<0.05),其中AEW组活菌比例下降幅度最小,与SAEW和52.5 g/L次氯酸钠液处理组相比,各时间点均有统计学差异(P<0.05);而SAEW与52.5 g/L次氯酸钠液处理组相比,各时间点的活菌比例均无统计学差异(P>0.05)。结论:SAEW和AEW对粪肠球菌均有杀菌作用,其中SAEW的作用与52.5 g/L次氯酸钠液相当;而AEW的杀菌效果较弱,且需要延长作用时间。     

饥饿期粪肠球菌生物膜毒力因子明胶酶E表达研究

目的:研究粪肠球菌生物膜形成相关毒力因子明胶酶E(gelE)在饥饿期及药物作用后的表达情况。方法:体外建立对数期、稳定期、饥饿期粪肠球菌生物膜模型,分别以1%、2.5%、5.25%次氯酸钠溶液作用于各时期粪肠球菌生物膜后,用Real-time PCR对gelE的基因表达相对量进行检测。结果:gelE基因表达相对量,饥饿期高于稳定期及对数期(P<0.05);用药后,3个期gelE基因表达相对量,5.25%次氯酸钠溶液组低于2.5%次氯酸钠溶液组和1%次氯酸钠溶液组(P<0.05)。结论:饥饿期粪肠球菌生物膜形成相关毒力因子gelE较对数期及稳定期表达增强。次氯酸钠浓度依赖性可抑制粪肠球菌毒力因子gelE的表达。     

茶多酚、MTAD和次氯酸钠液抑制粪肠球菌及其生物膜的作用比较

目的:检测中药茶多酚、MTAD、52.5 g/L次氯酸钠抑制根管壁粪肠球菌生物膜的能力,并分析中药茶多酚作为根管冲洗液的可行性。方法:采用微孔板滴定法检测茶多酚,MTAD和52.5 g/L次氯酸钠液对粪肠球菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),采用琼脂扩散实验检测3种抗菌剂对粪肠球菌的抑菌圈。通过建立粪肠球菌感染根管3周和6周的模型,分别用茶多酚、MTAD和52.5 g/L次氯酸钠液对粪肠球菌感染后的根管进行处理,然后对根管壁生物膜中的细菌生存情况进行定性定量分析,比较这3种根管冲洗液抗粪肠球菌生物膜的能力。结果:MIC、MBC和琼脂扩散实验显示:3组冲洗剂对粪肠球菌均有抑制作用。在粪肠球菌感染根管3周的生物膜中,MTAD和次氯酸钠液可以完全抑制粪肠球菌生物膜,茶多酚处理后虽仍有粪肠球菌生长,但明显低于生理盐水处理组的细菌生存数,差异有统计学意义(P<0.05)。在粪肠球菌感染根管6周的生物膜中,次氯酸钠液可以完全抑制细菌生长,茶多酚和MTAD作用后仍有活菌生长,但与对照组相比,细菌生存数均显示了8个log值的下降,差异有统计学意义(P<0.05)。结论:52.5 g/L次氯酸钠液具有很强的抑制根管内粪肠球菌生物膜作用,而茶多酚和MTAD也具有一定的抑制粪肠球菌生物膜的能力。     

次氯酸钠溶液抗菌体积分数的测定

目的评价次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌牙髓病原菌的体外抗菌性能。方法采用琼脂稀释法测定次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌的抗菌活性,其抗菌活性以最低抑菌体积分数(MIVF)和最低杀菌体积分数(MBVF)表示。结果次氯酸钠溶液对牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌和粪肠球菌的MIVF分别为0.035%、0.07%、0.018%和0.28%,MBVF分别为0.035%、0.07%、0.035%和0.28%,其总的抗菌效果排序分别为具核梭杆菌、牙龈卟啉单胞菌、中间普雷沃菌和粪肠球菌。结论次氯酸钠对口腔常见厌氧菌有抑制和杀灭作用,具有理想的抗菌性能。0.5%MBVF的次氯酸钠溶液的抑菌性能可满足临床需求。     

PIPS联合纳米银对根管粪肠球菌抗菌效果研究

目的 探究激光引发光声流系统(PIPS)和纳米银(AgNPs)联合使用对根管内粪肠球菌生物膜的抗菌效果。方法 收集36颗单根管离体牙,建立粪肠球菌感染根管实验模型,将样本随机分成6组,采用0.9%NaCl、2%NaClO、0.1%AgNPs溶液分别联合传统手动冲洗(CNI)或PIPS对根管进行冲洗,使用菌落计数法测定治疗前后根管内粪肠球菌生物膜菌落数,并计算菌落计数减少的百分比。结果 所有实验组粪肠球菌生物膜的抑制效果均强于对照组(P<0.05),使用PIPS辅助0.9%NaCl、2%NaClO、0.1%AgNPs冲洗组的降幅均分别大于CNI辅助0.9%NaCl、2%NaClO、0.1%AgNPs冲洗组(P<0.05)。PIPS辅助0.1%AgNPs冲洗组的降幅明显大于PIPS辅助2%NaClO冲洗组(P<0.05)。结论 PIPS辅助AgNPs溶液冲洗可以显著提高根管内粪肠球菌生物膜的清除效果。     

超声下不同根管冲洗剂组合对感染根管内粪肠球菌杀灭效果的研究

目的:比较超声作用下不同根管冲洗剂组合对根管内粪肠球菌的杀灭效果。方法:取粪肠球菌感染根管模型样本90个,随机分为6组(n=15),其中5个样本用于基线对照,10个样本分别按相应分组处理,A组:25 g/L次氯酸钠液液+20 g/L洗必泰液;B组:25 g/L次氯酸钠液+30 m L/L过氧化氢液;C组:10 g/L次氯酸钠液+20 g/L洗必泰液;D组:10 g/L次氯酸钠液+30 m L/L过氧化氢液;E组:52.5 g/L次氯酸钠液阳性对照;F组:生理盐水阴性对照。处理前后从样本的根管壁表面采样,并钻取不同深度牙本质进行细菌培养和计数。结果:A、B、C、D、E组根管壁表面及不同深度牙本质的细菌清除率均显著高于F组(P<0.05);对各层牙本质的细菌清除率,A组高于B、C、D组(P<0.05),与E组相比,除200~300μm深度处明显降低(P<0.05)外,其他各层与E组相当(P>0.05);C组对不同深度牙本质层的细菌清除率均在90%以上,低于A、E组,高于其他各组(P<0.05);B、D组在各不同深度牙本质层的细菌清除率基本相当(P>0.05),低于其他各实验组和E组(P<0.05)。结论:超声辅助下用25 g/L次氯酸钠液联合20 g/L洗必泰液冲洗根管,能有效杀灭根管内的粪肠球菌。     

Er:YAG激光联合次氯酸钠对粪肠球菌杀菌效果的研究

目的:比较掺铒钇铝-石榴石脉冲(Er:YAG)激光联合不同浓度次氯酸钠对根管内粪肠球菌杀菌效果的研究。方法:选择新鲜拔除的人单根管离体前磨牙建立粪肠球菌根管感染模型75个,随机分为5组,每组15个,分别进行如下处理:A组:5.25%次氯酸钠单独冲洗;B组:Er:YAG激光+5.25%次氯酸钠冲洗;C组:1%次氯酸钠单独冲洗;D组:Er:YAG激光+1%次氯酸钠冲洗。E组:空白对照。采集5组根管内的细菌进行培养计数并对结果进行卡方检验(Chi-square test)。结果:A、B、D三组均能完全杀灭根管内的粪肠球菌;C组未能完全杀灭根管内的粪肠球菌。15个培养皿中有11个可见粪肠球菌菌落。A、B、D三组间杀菌效果无差异且均显著优于C组。结论:高浓度次氯酸钠可以完全杀灭根管内的粪肠球菌,低浓度次氯酸钠不能完全杀灭根管内的粪肠球菌;Er:YAG激光可以显著增强低浓度次氯酸钠的杀菌作用。     

胞外多糖对粪肠球菌单菌种生物膜形成的影响

目的:观察粪肠球菌单菌种生物膜的动态形成过程,探讨胞外多糖在粪肠球菌生物膜形成过程中的分布和作用。方法:粪肠球菌在盖玻片上形成单菌种生物膜,采用荧光技术标记胞外多糖和细菌,荧光显微镜观察生物膜结构和胞外多糖的分布;蒽酮法测定粪肠球菌在游离和粘附状态下产生水溶性胞外多糖(Water soluble exopolysaccharides,WSE)和水不溶性胞外多糖(Water insoluble exopolysaccharides,WIE)的能力变化。结果:粪肠球菌能在玻片表面粘附形成生物膜,生物膜中胞外多糖分布与菌落中的细菌分布一致,相互包裹成熟形成网络状结构。不同培养时间、不同生存状态粪肠球菌合成WSE和WIE的能力不同,差异有显著性(P<0.05)。粘附菌产生WSE的能力往往低于浮游菌,但是粘附菌产生WIE均高于浮游菌,差异具有统计学意义(P<0.05)。结论:胞外多糖在粪肠球菌生物膜形成过程中发挥重要作用。     

不同pH值次氯酸钠对粪肠球菌抗菌效果的体外研究

目的：研究不同pH值2.5%次氯酸钠对粪肠球菌的抗菌效果。方法：采用纸片扩散法药敏实验观察pH12（A组）、pH9（B组）、pH7.5（C组）、pH6（D组）4种不同pH值2.5%次氯酸钠溶液和生理盐水（E组）对粪肠球菌的的抗菌效果,并分别测量其抑菌环直径的大小（mm）。结果：5组抑菌环直径从小到大依次为E组、A组、B组、C组和D组,其中A组与B组、A组与C组、B组与C组间差异均无显著性,D组与A组、B组、C组间差异均有显著性（P（0.01）,E组与A组、B组、C组、D组间差异均有显著性（P（0.01）。结论：随着pH值的降低,2.5%次氯酸钠溶液抑制粪肠球菌的效果逐渐增强,且pH为6时抗菌作用最强。     

Influence of Endodontic Procedure on the Adherence of Enterococcus faecalis

IntroductionThe purpose of this study was to investigate the effects of instrumentation and irrigation on the initial adherence of Enterococcus faecalis to root canal dentin and to explore initial microbial adhesion to root filling materials.MethodsThe following specimens were prepared: instrumented and uninstrumented dentin, dentin treated with different irrigation protocols, and root filling materials. The number of E. faecalis cells adhered on dentin was measured. The adhesion force of E. faecalis cells on different materials and the roughness of different surfaces were measured. The contact angle of the surfaces was recorded. The results were analyzed using the t test.ResultsInstrumented dentin specimens had a significantly higher amount of E. faecalis adherence than uninstrumented dentin. There were higher numbers of adhering bacteria on the dentin when EDTA was used alone (P < .05) compared with other irrigants alone. The use of chlorhexidine (CHX) as the last irrigant for a certain time resulted in a reduced number of adhering bacteria when the specimens were first exposed to sodium hypochlorite (NaOCl) followed by EDTA. EDTA used alone had the highest adhesion force followed by NaOCl alone and CHX alone (P < .05). Dentin treated with EDTA alone had the highest roughness and contact angle followed by NaOCl alone and CHX alone (P < .05). CHX added as the final irrigant after NaOCl with EDTA irrigation reduced the contact angle (P < .05). Larger amounts of adhering bacteria and higher adhesion force were detected on the surface of gutta-percha and sealer than on the dentin surface (P < .05).ConclusionsInstrumentation and irrigation alter the initial adherence of E. faecalis to root canal dentin and the surface properties of the dentin as well.     

Influence of irrigation regimens on the adherence of Enterococcus faecalis to root canal dentin

Enterococcus faecalis is frequently associated with post-treatment endodontic infections. Because adherence of bacteria to a substrate is the earliest stage in biofilm formation, eliciting the factors that links adherence of this bacterium to dentin would help in understanding its association with treatment-failed root canals. This investigation aimed to study the effects of endodontic irrigants on the adherence of E. faecalis to dentin. The bacteria adherence assay was conducted by using fluorescence microscopy, and the adhesion force was measured by using atomic force microscopy. There were significant increases in adherence and adhesion force after irrigation of dentin with ethylenediaminetetraacetic acid (EDTA), whereas sodium hypochlorite (NaOCl) reduced it. With the use of chlorhexidine (CHX), the force of adhesion increased, but the adherence assay showed a reduction in the number of adhering bacteria. The irrigation regimen of EDTA, NaOCl, and CHX resulted in the least number of adhering E. faecalis cells. This study highlighted that chemicals that alter the physicochemical properties of dentin will influence the nature of adherence, adhesion force, and subsequent biofilm formation of E. faecalis to dentin.     

Effect of smear layer and chlorhexidine treatment on the adhesion of Enterococcus faecalis to bovine dentin

The aim of this in vitro study was to determine the effects of a smear layer and chlorhexidine (CHX) treatment on the adhesion of Enterococcus faecalis to bovine dentin. Forty dentin blocks from bovine incisors were prepared and randomly divided into four groups of 10 each. The blocks in group 1 were placed in sterile saline for 5 minutes, while those in group 2 were treated with 17% EDTA for 5 minutes. The blocks in group 3 were placed in 2% CHX for 7 days. The blocks in group 4 were treated with 17% EDTA for 5 minutes, and then placed in 2% CHX for 7 days. All the blocks were immersed in a suspension of E. faecalis for 3 hours. The bacteria adhering to the dentin surface were counted by examination using a scanning electron microscope. The most significant amount of bacteria was retained on the samples from group 1 (p < 0.05) and the smallest amount of bacteria adhered to the samples from group 4. These results suggest that a smear layer enhances the adherence of E. faecalis to the dentin, and CHX is effective in reducing the adherence of microorganisms.     

Disinfection by endodontic irrigants and dressings of experimentally infected dentinal tubules

The effect of endodontic irrigants and dressings was tested on bacteria in bovine dentin specimens experimentally infected with Enterococcus faecalis, Streptococcus sanguis, Escherichia coli, or Pseudomonas aeruginosa. Standardized, cylindrical dentin test pieces were prepared and cleaned by ultrasonic treatment with EDTA and sodium hypochlorite. The specimens were infected with the test organism for periods up to 14 days, and the degree of infection into the tubules was monitored using Brown & Brenn stain, scanning electron microscopy, and culturing of dentin dust from sequential bur samples starting from the pulpal side. E. faecalis rapidly infected the whole length of the tubules; S. sanguis required up to 2 weeks for complete infection; E. coli only penetrated to some 600 microns, even after prolonged incubation periods. P. aeruginosa infected dentin quickly, but apparently in very low numbers. E. faecalis persisted for at least 10 d after withdrawal of nutrient support, whereas the other 3 organisms died within 4 to 48 h. Endodontic medicaments were applied to infected specimen for comparison of antibacterial potency. Camphorated p-monochlorophenol was generally more efficient than Calasept, and of the irrigants tested, iodine potassium iodide appeared more potent than sodium hypochlorite or chlorhexidine. The presence of a smear layer delayed, but did not eliminate, the effect of the medicaments.     

Effectiveness of different chemical agents for disinfection of gutta-percha cones

This aim of this study was to evaluate and compare the efficacy of different chemical methods to disinfect gutta-percha cones (GP). Eighty-six size 80 GP cones were used. The cones were contaminated by immersion in saliva and Enterococcus faecalis. Four chemical agents were used: 1% sodium hypochlorite (G1), 2% chlorhexidine gluconate (G2), 10% povidone iodine (G3) and 0.9% saline solution (G4). GP cones were immersed in the solutions for periods of 1 and 10 min. After the disinfection procedure, the cones were incubated in blood heart infusion and the presence of bacterial growth was analysed by turbidity of the medium. In G4, bacterial growth was observed in all specimens; G3 showed growth after immersion for 1 min when contaminated with E. faecalis; G1 showed diverse results after the immersion for 1 min. Meanwhile, G1 and G3 after 10 min, and G2 at both times evaluated did not show bacterial growth. The immersion of GP cones in 2% chlorhexidine gluconate for 1 min was an effective method for GP disinfection, while 10% povidone iodine and 1% sodium hypochlorite needed 10 min of immersion to disinfect the GP.     

Extension of Bactericidal Effect of Sodium Hypochlorite into Dentinal Tubules

Introduction

The aim of this study was to determine the extent of disinfection mediated by 2 concentrations of sodium hypochlorite for bacteria-laden dentin wall of a root canal.

Methods

A dual-species film, Enterococcus faecalis and Porphyromonas gingivalis, was cultivated on root canal dentin for 7 days. The canal wall was irrigated with 0.5% or 3% sodium hypochlorite (n = 8 each), with a control group (n = 2) irrigated with sterile saline. The roots were then split horizontally at 5-, 7-, and 9-mm levels from root apex and examined under confocal laser scanning microscope and then scanning electron microscope. The proportions of viable cells situated on different depths into root canal dentin were compared at a significance level of P = .05.

Results

Both concentrations of sodium hypochlorite significantly reduced the amount of live bacteria in the most superficial layer (first 0.1 mm) of root canal dentin, compared with the control; however, the difference was not significant between the 2 concentrations. For the next 2 layers (0.1–0.3 mm into dentinal tubules), irrigation with 3% hypochlorite resulted in significantly lower amounts of viable bacteria than 0.5% hypochlorite or saline. No significant difference between 0.5% or 3% sodium hypochlorite and the positive control was observed in deeper (>0.3 mm) regions into the root canal wall.

Conclusions

Increasing the concentration of sodium hypochlorite improves the penetration depth of its antibacterial action into dentinal tubules, but that seemed unable to completely eradicate bacteria residing there. Total elimination of bacteria from dentinal tubules cannot be achieved by irrigation with sodium hypochlorite alone.     

Tissue-dissolving capacity and antibacterial effect of buffered and unbuffered hypochlorite solutions

OBJECTIVE: The goal of this study was to compare the dissolving potential of Dakin's solution with that of equivalent buffered and unbuffered sodium hypochlorite solutions on fresh and decayed tissues. In addition, the antimicrobial effect of Dakin's solution and equivalent unbuffered hypochlorite was tested. STUDY DESIGN: Tissue specimens were obtained from freshly dissected pig palates. Unbuffered 2.5% and 0.5% sodium hypochlorite solutions and 0.5% solutions buffered at a pH of 12 and a pH of 9 (Dakin's solution) were tested on fresh and decayed tissue. Tissue decay was assessed histologically. Antimicrobial testing was performed with Enterococcus faecalis in dentin blocks and on filter papers. RESULTS: The 2.5% NaOCl solution was substantially more effective than the three 0.5% solutions in dissolving the test tissues. Buffering had little effect on tissue dissolution, and Dakin's solution was equally effective on decayed and fresh tissues. No differences were recorded for the antibacterial properties of Dakin's solution and an equivalent unbuffered hypochlorite solution. CONCLUSIONS: In contrast to earlier statements, the results of this study do not demonstrate any benefit from buffering sodium hypochlorite with sodium bicarbonate according to Dakin's method. An irrigation solution with less dissolving potential may be obtained by simply diluting stock solutions of NaOCl with water.     

Pilot study of conditioner/primer effects on resin-dentin bonding after provisional cement contamination using SEM, energy dispersive x-ray spectroscopy, and bond strength evaluation measures

STATEMENT OF PROBLEM: Previous studies have shown that provisional cement cannot be completely removed from the dentin surface and the remnants inhibit adhesion of resin cement to dentin. PURPOSE: This study investigated the effects of dentin conditioners on resin-dentin bonding after provisional cement contamination. MATERIAL AND METHODS: A total of 216 bovine dentin specimens were divided into 2 groups with and without provisional cement coating on the dentin surface. Each group was further divided into 3 subgroups with 2 conditioning methods and without dentin conditioning (36 specimens per subgroup). The 2 methods applied were (1) phosphoric acid etching and (2) phosphoric acid etching, followed by sodium hypochlorite gel. Provisional cement was completely removed with a curette before dentin conditioning. Stainless steel rods were luted to dentin surfaces with Panavia 21 resin cement. Tensile bond strengths were measured before and after thermocycling. The dentin surface was analyzed by SEM and energy dispersive x-ray spectroscopy. Three-way ANOVA and Scheffe's comparison test were performed (P =. 05). RESULTS: Bond strength was significantly influenced by provisional cement application, type of dentin conditioning, and thermocycling. Bond strength decreased when treated with provisional cement, but the decreased strengths were restored to their original values or higher by both dentin conditioners used. Specimens treated with sodium hypochlorite gel applied after phosphoric acid etching demonstrated the highest bond strengths with good stability after thermocycling. CONCLUSION: Application of sodium hypochlorite gel after phosphoric acid etching was more effective than etching with phosphoric acid alone to eliminate the inhibitory effect of provisional cement remnants on adhesion between resin cement and dentin.