OBJECTIVETo evaluate in vitro the antibacterial effectiveness of sodium hypochlorite on Enterococcus faecalis (E. faecalis) within root canals.

METHODSRoots inoculated with E. faecalis were divided into six groups, which underwent different root canal irrigation and treatment: Group 1: 5.25% NaOCl, Group 2: 2.5% NaOCl, Group 3: 0.9% NaCl, Group 4: root canal preparation + 5.25% NaOCl, Group 5: root canal preparation + 2.5% NaOCl, Group 6: root canal preparation + 0.9% NaCl. Microbiological samples were collected from root canals at three time points (before irrigation, immediately after irrigation and three days after irrigation).

RESULTSThe numbers of E. faecalis in root canal in each of the six groups were effectively reduced. Group 1 and 2 were statistically more effective than Group 3. Group 4 was statistically more effective than Group 5 and 6. Group 5 was statistically more effective than Group 6. Bacterial survival was observed in all canals of each group after a 3-day-incubation.

CONCLUSIONS2.5% NaOCl was highly effective for root canal irrigation. However residual bacteria remained in the root canals after chemical irrigation by NaOCl and mechanical preparation.

Dental Pulp Cavity ; microbiology ; Enterococcus faecalis ; drug effects ; Humans ; In Vitro Techniques ; Root Canal Irrigants ; pharmacology ; Sodium Hypochlorite ; pharmacology

OBJECTIVETo compare the antimicrobial efficacy of four endodontic irrigants using an in vitro model infected by Enterococcus faecalis (Ef).

METHODSThe root canals of fifty extracted teeth were infected by Ef in vitro. The test groups were irrigated with 3% H(2)O(2), 2.5% sodium hypochlorite (SH), 2% chloramine-T (CR), and 2% chlorhexidine (CHX), respectively, and the control group was irrigated with 0.9% NaCl. The concentration of Ef in canals of each group was calculated before and after irrigation. The residual bacteria within the dentinal tubules and vitalities of the residual bacteria were also examined.

RESULTSAll chemical irrigants were significantly more effective than 0.9% NaCl (P < 0.05); 2.5% SH and 2% CHX were statistically more effective than 3% H(2)O(2) (P < 0.05). Residual bacteria could be found in the dentinal tubules and propagated 72 h after.

CONCLUSIONS2% CR and 2% CHX had almost the equivalent antimicrobial effect as 2.5% SH, but 3% H(2)O(2) was less effective.

Chloramines ; pharmacology ; Chlorhexidine ; pharmacology ; Dental Pulp Cavity ; microbiology ; Enterococcus faecalis ; drug effects ; Humans ; Hydrogen Peroxide ; pharmacology ; In Vitro Techniques ; Root Canal Irrigants ; pharmacology ; Sodium Hypochlorite ; pharmacology ; Tosyl Compounds ; pharmacology

OBJECTIVETo investigate the effects of putative bacteriocin immunity proteins on the growth mode of Streptococcus mutans (Sm). To observe the differences of antimicrobial sensitivity in planktonic Sm wild-type strains and mutant strains caused by the inactivation of bacteriocin immunity proteins and their influence on the biofilm formation.

METHODSSm wild-type strains (WT) and its knockout mutants defective in immA and immB (ΔimmA(-) and ΔimmB(-) mutants) coding putative bacteriocin immunity proteins were cultured in brain heart infusion (BHI) and selected by erythromycin at the concentration of 10 mg/L. Optical density was detected by spectrophotometer every hour and growth curve was drawn. WT, ΔimmA(-) and ΔimmB(-) mutants were treated with ampicillin (0.04, 0.05, 0.06, 0.07, 0.08 mg/L), sodium fluoride (50, 100, 150, 200, 250 mg/L) and sodium hypochlorite (0.078%, 0.156%, 0.313%, 0.625%, 1.250%) for 24 hours. Optical density was detected by multifunctional micro plate reader. WT and the mutants were cultured in MBEC(TM) P&G Assay for 24 hours. The minimum biofilm eradication concentration (MBEC) of chlorhexidine against Sm was determined by serial dilution method. Confocal laser scanning microscopy (CLSM) was used to visualize the biofilm architecture, depth and ratio of live to dead bacteria.

RESULTSGrowth curve showed that it took about 3 hours to reach exponential phase and about 7 hours to stationary phase for WT, while 4 hours to exponential phase and 8 hours to stationary phase for mutants. Optical density of mutants were lower than WT in the presence of various antimicrobial agents (P < 0.01). In 0.06 mg/L ampicillin group, optical density value of WT, ΔimmA(-) and ΔimmB(-) mutants were 0.334 ± 0.016, 0.027 ± 0.016 and 0.047 ± 0.018. In 150 mg/L sodium fluoride group, optical density value of WT and mutants were 0.254 ± 0.018, 0.129 ± 0.011 and 0.167 ± 0.010. In 0.313% sodium hypochlorite group, optical density value of WT and mutants were 0.467 ± 0.008, 0.017 ± 0.006 and 0.050 ± 0.006. The MBEC of chlorhexidine against Sm WT, ΔimmA(-) and ΔimmB(-) mutants were 6.25, 1.57, and 3.13 mg/L. The results by CLSM showed a noticeable difference in biofilm architecture. The depth of WT biofilm was higher than the mutants biofilm (P < 0.01). The ratio of live to dead bacteria of WT biofilm was higher than ΔimmA(-) mutants in all layers (P < 0.05) and ΔimmB(-) mutants in the outer and intermedium layer (P < 0.01). There is no significant different between the inner layers of WT and ΔimmB(-) mutants (P = 0.191).

CONCLUSIONSPutative bacteriocin immunity proteins have influence on the growth mode of Sm. The antimicrobial sensitivity of planktonic Sm can be up-regulated by the inactivation of immA or immB. The MBEC of chlorhexidine against ΔimmA(-) and ΔimmB(-) mutants is lower than WT. The inactivation of immA or immB affects the biofilm formation.

Ampicillin ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacteriocins ; genetics ; immunology ; Biofilms ; drug effects ; growth & development ; Cariostatic Agents ; pharmacology ; Chlorhexidine ; pharmacology ; Disinfectants ; pharmacology ; Microbial Sensitivity Tests ; Mutation ; Plankton ; drug effects ; Sodium Fluoride ; pharmacology ; Sodium Hypochlorite ; pharmacology ; Streptococcus mutans ; drug effects ; genetics

OBJECTIVETo scan the most resistable bacteriophage as an indicator in disinfection tests, and to study the resistance of bacteriophage T4, Phichi 174D, and f2 to the sodium dichloroisocyanurate (NaDCC) in laboratory.

METHODSThe virucidal activity of NaDCC against bacteriophage T4, Phichi 174D, and f2 were assessed by suspension test. The neutralizer was selected and be appraised by test of neutralizer. Bacteriophage T4, Phichi 174D, and f2 were detected and enumerated by the double-agar-layer plaque technique.

RESULTS(1) With 150 mg/L of available chlorine of NaDCC solution, within a contact time of 40 minutes, or 300 mg/L, 5 minutes, the reductions of bacteriophage T4 achieved the "disinfection" level [log(10) inactivation value or log(10) reduction value of bacteriophage T4 (log(10)No-log(10)Nt) > or = 4.00 log(10)]. (2) With 300 mg/L of available chlorine of NaDCC solution, within a contact time of 5 minutes, or 400 mg/L, 3 minutes, the reductions of bacteriophage Phichi 174D achieved the "disinfection" level. (3) With 2000 mg/L of available chlorine of NaDCC solution, within a contact time of 20 minutes, or 4000 mg/L, 5 minutes, the reductions of bacteriophage f2 might achieve the "disinfection" level.

CONCLUSIONThe order of resistance of the above three bacteriophages to NaDCC from greatest to smallest is as follows: bacteriophage f2 > bacteriophage T4 > bacteriophage Phichi 174D.

Bacteriophage T4 ; drug effects ; Bacteriophage phi X 174 ; drug effects ; Bacteriophages ; drug effects ; Disinfectants ; pharmacology ; Drug Resistance, Viral ; Sodium Hypochlorite ; pharmacology

OBJECTIVETo study the changes of growth and biofilm formation capability of Enterococcus faecalis (Ef) in different stress conditions.

METHODSThe changes of growth of Ef in stress conditions were observed by measuring the A600 value with ultraviolet spectrophotometer. Ef was incubated on glass slide in stress conditions, biofilm formation capability of cells was investigated by colony-forming unit (CFU) counting of the culturable bacteria and fluorescence confocal laser scanning microscopy.

RESULTSEf couldn't growth under the conditions of 2%, 5%NaClO, pH = 11 and 12, the A600 value was unchanged in 96 hours. But the growth curve changed at different levels in other stress conditions: under 1%NaClO, the A600 value peaked at 1.461 at 16 hour (the peaked level was 1.238 at 6 hours in control group) ; under 0,0.05%,0.15% glucose, it peaked at 0.645,0.890, 1.173, respectively, at 6 hour (it was maximized to 1.195 at 6 hours in control group); the A600 value peaked at 1.704 at 6 hours at pH = 9 and 1.225 at 10 hours at pH = 10 (the peak level was 1.732 at 6 hours at pH = 7) . Biofilm assay showed that Ef were able to form biofilm in these stress conditions except 5%NaClO and pH = 12.

CONCLUSIONSEf could growth and form biofilms in energy starvation, low concentrations of sodium hypochlorite and weak alkaline stress.

Biofilms ; drug effects ; growth & development ; Colony Count, Microbial ; Enterococcus faecalis ; drug effects ; growth & development ; Glucose ; pharmacology ; Hydrogen-Ion Concentration ; Microscopy, Confocal ; Sodium Hypochlorite ; pharmacology

OBJECTIVES: Root canal therapy is the most effective and common method for pulpitis and periapical periodontitis. During the root canal preparation, chemical irrigation plays a key role. However, sodium hypochlorite (NaOCl), the widely used irrigation fluid, may impact the bonding strength between dentin and restorative material meanwhile sterilization and dissolving. Therefore, it's important to explore the influence of NaOCl on the adhesion between dentin and restoration materials to ensure clinical efficacy. This study aims to explore the effect of NaOCl on dentine adhesion and evaluate the effect of dentine adhesion induced by sodium erythorbate (ERY), and to provide clinical guidance on dentin bonding after root canal therapy. METHODS: Seventy freshly complete extracted human third molars aged 18-33 years old, without caries and restorations were selected. A diamond saw was used under running water to achieve dentine fragments which were divided into 10 groups with 14 fragments in each group: 2 control [deionized water (DW)±10% ERY] and 8 experimental groups (0.5%, 1%, 2.5%, and 5.25% NaOCl±10% ERY). The dentine specimens in the control group (treated with DW) and the experimental groups (treated with 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl) were immersed for 20 min using corresponding solutions which were renewed every 5 min. The other 5 groups were immersed in 10% ERY for 5 min after an initial washing with DW for 1 min. Then, we selected 4 dentine fragments from all 14 fragments in each group and the numbers and diameters of opening dentinal tubules were observed under scanning electron microscope (SEM). The other 10 dentine fragments from each group were used to make adhesive samples by using self-etch adhesive wand composite resin. All the above adhesive samples were sectioned perpendicular to the bonded interface into 20 slabs with a cross-sectional area of 1 mm×1 mm using a diamond saw under the cooling water, and then the morphology of 10 slabs in each group's bonding interface was observed from aspects of formation of resin tags, depth of tags in dentin, and formation of hybrid layer under SEM. The other 10 slabs of each group's microtensile bond strength and failure modes were also analyzed. RESULTS: Among the 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules gradually increased with the rise of concentration of NaOCl solution (all P<0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules increased after using ERY, but without significant difference (all P>0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, the scores of formation of resin tags under SEM gradually increased with the increase of concentration of NaOCl solution, while the score in the 5.25% NaOCl group decreased significantly compared with the score of the 2.5% NaOCl group (P<0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (all P>0.05). The scores of length of the tags under SEM in the 5.25% NaOCl group was significantly higher than the scores of DW, 0.5% NaOCl, and 1% NaOCl groups (all P<0.05), and it was also higher than the score of the 2.5% NaOCl group, but without significant difference (P>0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (P>0.05). The scores of formation of hybrid layer under SEM in the 2.5% NaOCl and 5.25% NaOCl groups significantly decreased compared with the score of the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). Microtensile bond strength was greater in the 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, but lower in the 5.25% NaOCl group than that in the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). The incidence of type "Adhesive" of failure modes in the 5.25% NaOCl group was significantly higher than that in other groups (all P<0.05), while the incidence of type "Adhesive" in the 5.25% NaOCl+10% ERY group was lower than that in the 5.25% NaOCl group (P<0.05). CONCLUSIONS The bonding strength to dentine increases with the increase of NaOCl concentration when the concentration lower than 2.5%; whereas it is decreased at a higher concentration (such as 5.25%). 10% ERY has a definite recovery effect on attenuated bonding strength to 5.25% NaOCl-treated dentine.
Adolescent ; Adult ; Ascorbic Acid ; Dental Bonding ; Dentin ; Dentin-Bonding Agents/pharmacology* ; Diamond/pharmacology* ; Humans ; Materials Testing ; Microscopy, Electron, Scanning ; Resin Cements/pharmacology* ; Sodium Hypochlorite/pharmacology* ; Tensile Strength ; Water/pharmacology* ; Young Adult