scholarly journals Use of electromagnetic stimulation on an Enterococcus faecalis biofilm on root canal treated teeth in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Beatriz H. D. Panariello ◽  
Justin K. Kindler ◽  
Kenneth J. Spolnik ◽  
Ygal Ehrlich ◽  
George J. Eckert ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Imaging ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Microscopy Imaging ◽  
Electromagnetic Stimulation ◽  
Root Canal Disinfection

AbstractRoot canal disinfection is of utmost importance in the success of the treatment, thus, a novel method for achieving root canal disinfection by electromagnetic waves, creating a synergistic reaction via electric and thermal energy, was created. To study electromagnetic stimulation (EMS) for the disinfection of root canal in vitro, single rooted teeth were instrumented with a 45.05 Wave One Gold reciprocating file. Specimens were sterilized and inoculated with Enterococcus faecalis ATCC 29,212, which grew for 15 days to form an established biofilm. Samples were treated with 6% sodium hypochlorite (NaOCl), 1.5% NaOCl 1.5% NaOCl with EMS, 0.9% saline with EMS or 0.9% saline. After treatments, the colony forming units (CFU) was determined. Data was analyzed by Wilcoxon Rank Sums Test (α = 0.05). One sample per group was scored and split for confocal laser scanning microscopy imaging. There was a significant effect with the use of NaOCl with or without EMS versus 0.9% saline with or without EMS (p = 0.012 and 0.003, respectively). CFUs were lower when using 0.9% saline with EMS versus 0.9% saline alone (p = 0.002). Confocal imaging confirmed CFU findings. EMS with saline has an antibiofilm effect against E. faecalis and can potentially be applied for endodontic disinfection.

scholarly journals Effectiveness of chitosan-propolis nanoparticle against Enterococcus faecalis biofilms in the root canal

2020 ◽  
Author(s):  
Abhishek Parolia ◽  
Haresh Kumar Kumar ◽  
Srinivasan Ramamurthy ◽  
Allan Pau
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Internal Diameter ◽  
Group Vi ◽  
Human Teeth ◽  
Group V ◽  
Group I

Abstract Background To determine the antibacterial effect of chitosan-propolis nanoparticle (CPN) as an intracanal medicament against Enterococcus faecalis biofilm in root canal. Methods 240 extracted human teeth were sectioned to obtain 6mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into eight groups ( n=30 ) according to the intracanal medicament placed: group I: saline, groupII: chitosan, group III: propolis100 µg/ml (P100), group IV: propolis 250 µg/ml (P250), group V: chitosan-propolis nanoparticle 100µg/ml (CPN100), group VI: chitosan-propolis nanoparticle 250 µg/ml (CPN250), group VII: calcium hydroxide(CH) and group VIII: 2% chlorhexidine (CHX) gel. Dentine shavings were collected at 200 and 400 μm depths, and total numbers of CFUs were determined at the end of day one, three and seven. The non-parametric Kruskal Wallis and Mann-Whitney tests were used to compare the differences in reduction of CFUs between all groups and probability values of P < 0.05 were set as the reference for statistically significant results. The scanning electron microscope (SEM) and confocal laser scanning microscopy (CLSM) were also performed after exposure to CPNs. The effectiveness of CPNs were also evaluated against E. faecalis isolated obtained from patients having failed root canal treatment. Results Reduction in the number of colony‐forming units was statistically significant in all groups compared to saline (p <.05). On day one and three, at 200 and 400-μm, CPN250 showed significant reduction of CFUs compared to all other groups (p <.05), while CPN100 was significantly better than other groups (p <.05) except CPN250 and CHX. On day seven, at 200-μm CPN250 showed significant reduction of CFUs compared to all other groups (p <.05) except CPN100 and CHX, while at 400 μm CPN250 showed similar effectiveness as CPN100, CH and CHX. SEM and CLSM images also showed the maximum reduction of E. faecalis with CPN250. Conclusion CPN250 was the most effective in reducing E. faecalis colonies on day one, three at both depths and at day seven CPN250 was equally effective as CPN100 and CHX.

Microanalysis of Thermal-Induced Changes at the Resin–Dentin Interface

2014 ◽  
Vol 20 (4) ◽  
pp. 1218-1233 ◽  
Author(s):  
Manuel Toledano ◽  
Fátima S. Aguilera ◽  
Estrella Osorio ◽  
Inmaculada Cabello ◽  
Raquel Osorio
Keyword(s):  
Cluster Analysis ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Hybrid Layer ◽  
Microscopy Imaging ◽  
Etch And Rinse ◽  
Induced Changes ◽  
Dentin Adhesive

AbstractThe purpose of this study was to evaluate the ability of two dentin adhesive systems to induce remineralization in the bonded dentin interface afterin vitrothermo-cycling. Dentin surfaces were treated with two different adhesive approaches: (1) 37% phosphoric acid (PA) plus an “etch-and-rinse” dentin adhesive (single bond, SB) (PA+SB) or (2) application of a “self-etch” dentin adhesive (Clearfil SE bond, SEB). Three groups were established: (i) 24 h or (ii) 3 m storage, and (iii) specimens submitted to thermal cycling (100,000 cy/5 and 55ºC). Atomic force microscopy imaging/nanoindentation, Raman spectroscopy/cluster analysis with dye-assisted confocal laser scanning microscopy (CLSM) evaluation and Masson’s trichrome staining assessments were implemented for characterization. Thermo-cycling increased nanohardness in PA+SB at the hybrid layer (HL) and in SEB at the bottom of the HL (BHL). Young’s modulus increased at both the HL and BHL in SEB and at the HL in PA+SB, after thermal stress. Cluster analysis demonstrated an augmentation of the mineral–matrix ratio in thermo-cycled specimens. CLSM showed a decrease of both micropermeability and nanoleakage after thermo-cycling in PA+SB, and were completely absent in SEB. Trichrome staining reflected a scarce demineralized front in PA+SB after thermo-cycling and total remineralization in SEB.

scholarly journals Evaluation of Antibacterial Efficacy of Vitex negundo Linn. extract as Root Canal Irrigant against Enterococcus faecalis and its penetration into Root Dentin: An in-vitro study

2020 ◽  
Vol 11 (2) ◽  
pp. 193-199
Author(s):  
Suruchi Santosh Gupta ◽  
Nilima Thosar ◽  
Nilesh Rathi ◽  
Sudhindra M Baliga ◽  
Yagnesh Thakkar ◽  
...  
Keyword(s):  
Penetration Depth ◽  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Antibacterial Efficacy ◽  
Vitex Negundo ◽  
Root Dentin ◽  
Root Canal Irrigant

Introduction: The aim of this study was to evaluate the antibacterial efficacy of Vitex negundo Linn. extract as root canal irrigant against Enterococcus faecalis and its penetration into root dentin. Methods and Materials: Forty single rooted premolars were randomly divided into 4 groups: 3% Sodium hypochlorite (NaOCl), 2% Chlorhexidine (CHX) , 100mg/ml Vitex negundo Linn. and saline as control all mixed with Rhodamine B dye. Test samples were analysed for bacterial count before and after irrigation using absorbent paper points and the colony forming units were recorded and measured. Sectioning of the samples was performed at three levels 3mm,6mm,9mm from apex and then these samples were analysed using confocal laser scanning microscopy for penetration depth of the irrigant within the dentinal tubules. Paired t-test and ANOVA test were used to perform statistical analysis with level of significance set at 0.05 Results: The mean CFU/ml count of Enterococcus facealis reduced significantly in all the groups post irrigation. All the irrigants showed maximum penetration depth at coronal third level compared to middle and apical third level respectively. The penetration depth of NaOCl group was better when compared to CHX group and Vitex negundo Linn. group but the difference was statistically not significant. Conclusion: Although 3% NaOCl was the most effective irrigant, all agents exerted acceptable antimicrobial activity against Enterococcus faecalis and penetration depth within tubules of dentin.

scholarly journals Effectiveness of chitosan-propolis nanoparticle against Enterococcus faecalis biofilms in the root canal

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Abhishek Parolia ◽  
Haresh Kumar ◽  
Srinivasan Ramamurthy ◽  
Fabian Davamani ◽  
Allan Pau
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Current Trend ◽  
Laser Scanning Microscopy ◽  
Successful Outcome ◽  
Confocal Laser ◽  
Internal Diameter ◽  
Sem Images ◽  
Group I

Abstract Background The successful outcome of endodontic treatment depends on controlling the intra-radicular microbial biofilm by effective instrumentation and disinfection using various irrigants and intracanal medicaments. Instrumentation alone cannot effectively debride the root canals specially due to the complex morphology of the root canal system. A number of antibiotics and surfactants are being widely used in the treatment of biofilms however, the current trend is towards identification of natural products in disinfection. The aim of the study was to determine the antibacterial effect of chitosan-propolis nanoparticle (CPN) as an intracanal medicament against Enterococcus faecalis biofilm in root canal. Methods 240 extracted human teeth were sectioned to obtain 6 mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9 mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into eight groups (n = 30) according to the intracanal medicament placed: group I: saline, group II: chitosan, group III: propolis100 µg/ml (P100), group IV: propolis 250 µg/ml (P250), group V: chitosan-propolis nanoparticle 100 µg/ml (CPN100), group VI: chitosan-propolis nanoparticle 250 µg/ml (CPN250), group VII: calcium hydroxide(CH) and group VIII: 2% chlorhexidine (CHX) gel. Dentine shavings were collected at 200 and 400 μm depths, and total numbers of CFUs were determined at the end of day one, three and seven. The non-parametric Kruskal Wallis and Mann–Whitney tests were used to compare the differences in reduction of CFUs between all groups and probability values of p < 0.05 were set as the reference for statistically significant results. The scanning electron microscope (SEM) and confocal laser scanning microscopy (CLSM) were also performed after exposure to CPNs. The effectiveness of CPNs were also evaluated against E. faecalis isolated obtained from patients having failed root canal treatment. Results The treatments of chitosan, P100, P250, CPN100, CPN250, CH and 2% CHX reduced the CFUs significantly compared to saline (p < .05). On day one and three, at 200 and 400-μm, CPN250 showed significant reduction of CFUs compared to all other groups (p < .05), while CPN100 was significantly better than other groups (p < .05) except CPN250 and 2% CHX. On day seven, at 200-μm CPN250 showed significant reduction of CFUs compared to all other groups (p < .05) except CPN100 and CHX, while at 400 μm CPN250 showed similar effectiveness as CPN100, CH and 2% CHX. SEM images showed root canal dentin treated with CPN250 had less coverage with E. faecalis bacteria similarly, CLSM images also showed higher percentage of dead E. faecalis bacteria with CPN250 than to CPN100. Conclusion CPN250 was the most effective in reducing E. faecalis colonies on day one, three at both depths and at day seven CPN250 was equally effective as CPN100 and 2% CHX.

scholarly journals Characterization of Enterococcus faecalis in different culture conditions

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Mi-Ah Kim ◽  
Vinicius Rosa ◽  
Kyung-San Min
Keyword(s):  
Enterococcus Faecalis ◽  
Laser Scanning ◽  
Colorimetric Assay ◽  
Culture Conditions ◽  
Laser Scanning Microscopy ◽  
Extracellular Dna ◽  
Confocal Laser ◽  
Qrt Pcr ◽  
Microscopy Analysis

AbstractThe aim of this study was to investigate how carbohydrates (glucose or sucrose) affect the characteristics of Enterococcus faecalis (E. faecalis) planktonic and biofilm in vitro. For this study, E. faecalis was cultured in tryptone-yeast extract broth with 0% glucose + 0% sucrose, 0.5% glucose, 1% glucose, 0.5% sucrose, or 1% sucrose. Viability of E. faecalis was examined by colony forming unit counting assays. Biofilm formation was assessed by measuring extracellular DNA (eDNA), a component of the biofilm matrix. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression of virulence-associated genes. Field emission scanning electron microscopy analysis, confocal laser scanning microscopy analysis, and crystal violet colorimetric assay were conducted to study E. faecalis biofilms. E. faecalis showed the highest viability and eDNA levels in 1% sucrose medium in biofilms. The result of qRT-PCR showed that the virulence-associated genes expressed highest in 1% sucrose-grown biofilms and in 1% glucose-grown planktonic cultures. E. faecalis showed highly aggregated biofilms and higher bacteria and exopolysaccharide (EPS) bio-volume in sucrose than in 0% glucose + 0% sucrose or glucose. The results indicate that the production of eDNA and EPS and expression of virulence-associated genes in E. faecalis are affected by the concentration of carbohydrates in biofilm or planktonic culture.

scholarly journals Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 715
Author(s):  
Abhishek Parolia ◽  
Haresh Kumar ◽  
Srinivasan Ramamurthy ◽  
Thiagarajan Madheswaran ◽  
Fabian Davamani ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Bioactive Compounds ◽  
Root Canal ◽  
Laser Scanning ◽  
Antibacterial Effect ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Internal Diameter ◽  
Group I ◽  
Significant Difference

To determine the antibacterial effect of propolis nanoparticles (PNs) as an endodontic irrigant against Enterococcus faecalis biofilm inside the endodontic root canal system. Two-hundred-ten extracted human teeth were sectioned to obtain 6 mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9 mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into seven groups, with 30 dentinal blocks in each group including: group I—saline; group II—propolis 100 µg/mL; group III—propolis 300 µg/mL; group IV—propolis nanoparticle 100 µg/mL; group V—propolis nanoparticle 300µg/mL; group VI—6% sodium hypochlorite; group VII—2% chlorhexidine. Dentin shavings were collected at 200 and 400 μm depths, and total numbers of CFUs were determined at the end of one, five, and ten minutes. The non-parametric Kruskal–Wallis and Mann–Whitney tests were used to compare the differences in reduction in CFUs between all groups, and probability values of p < 0.05 were set as the reference for statistically significant results. The antibacterial effect of PNs as an endodontic irrigant was also assessed against E. faecalis isolates from patients with failed root canal treatment. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were also performed after exposure to PNs. A Raman spectroscope, equipped with a Leica microscope and lenses with curve-fitting Raman software, was used for analysis. The molecular interactions between bioactive compounds of propolis (Pinocembrin, Kaempferol, and Quercetin) and the proteins Sortase A and β-galactosidase were also understood by computational molecular docking studies. PN300 was significantly more effective in reducing CFUs compared to all other groups (p < 0.05) except 6% NaOCl and 2% CHX (p > 0.05) at all time intervals and both depths. At five minutes, 6% NaOCl and 2% CHX were the most effective in reducing CFUs (p < 0.05). However, no significant difference was found between PN300, 6% NaOCl, and 2% CHX at 10 min (p > 0.05). SEM images also showed the maximum reduction in E. faecalis with PN300, 6% NaOCl, and 2% CHX at five and ten minutes. CLSM images showed the number of dead cells in dentin were highest with PN300 compared to PN100 and saline. There was a reduction in the 484 cm−1 band and an increase in the 870 cm−1 band in the PN300 group. The detailed observations of the docking poses of bioactive compounds and their interactions with key residues of the binding site in all the three docking protocols revealed that the interactions were consistent with reasonable docking and IFD docking scores. PN300 was equally as effective as 6% NaOCl and 2% CHX in reducing the E. faecalis biofilms.

Antibacterial effect of propolis nanoparticles against Enterococcus faecalis biofilm in the root canal

2019 ◽  
Author(s):  
Abhishek Parolia ◽  
Haresh Kumar Kumar ◽  
Srinivasan Ramamurthy Ramamurthy ◽  
Allan Pau
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Antibacterial Effect ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Internal Diameter ◽  
Sem Images ◽  
Group I ◽  
Significant Difference

Abstract Background: To determine the antibacterial effect of propolis nanoparticles (PNs) as an endodontic irrigant against Enterococcus faecalis biofilm in root canal. Methods: PNs were prepared by ultrasonication and the particle size distribution and polydispersity index were determined by dynamic light scattering using Zetasizer Nano S90. 210 extracted human teeth were sectioned to obtain 6mm of the middle third of the root. The root canal was enlarged to an internal diameter of 0.9mm. The specimens were inoculated with E. faecalis for 21 days. Following this, specimens were randomly divided into seven groups having 30 dentinal blocks in each group including group I: saline, group II: propolis 100µg/ml, group III: propolis 300µg/ml, group IV: propolis nanoparticle 100µg/ml, group V: propolis nanoparticle 300µg/ml, group VI: 6% sodium hypochlorite, group VII: 2% chlorhexidine. Dentine shavings were collected at 200 and 400 μm depths, and total numbers of CFUs were determined at the end of one, five, and ten minutes. The non-parametric Kruskal Wallis and Mann-Whitney tests were used to compare the differences in reduction of CFUs between all groups and probability values of P < 0.05 were set as the reference for statistically significant results. The scanning electron microscope and confocal laser scanning microscopy were also performed after exposure to PNs. Results: PN300 was significantly more effective in reducing CFUs compared to all other groups (p <0.05) except 6% NaOCl and 2% CHX (p >0.05) at all-time intervals and both depths. At five minutes, 6% NaOCl and 2 % CHX were the most effective in reducing CFUs (p <0.05) however, no significant difference was found in between PN300, 6% NaOCl and 2 % CHX at 10 minutes (p >0.05). SEM images also showed the maximum reduction of E. faecalis with PN300, 6% NaOCl and 2% CHX (>90 %) at five and ten minutes. CLSM images showed the number of dead cells in dentin was highest with PN300 (>90%) compared to PN100 (>40%) and saline (all live cells). Conclusion: PN300 was equally effective as 6% NaOCl, and 2% CHX in reducing E. faecalis CFUs after one minute, five and ten minutes at both depths.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

scholarly journals Effect of dissolution rate and subsequent ion release on cytocompatibility properties of borophosphate glasses

2019 ◽  
Vol 5 (1) ◽  
pp. 85-97
Author(s):  
Nusrat Sharmin ◽  
Mohammad S. Hasan ◽  
Md. Towhidul Islam ◽  
Chengheng Pang ◽  
Fu Gu ◽  
...  
Keyword(s):  
Dissolution Rate ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ion Release ◽  
Scanning Calorimetry ◽  
Cell Culture Study ◽  
Borophosphate Glasses ◽  
Mg63 Cells

AbstractPresent work explores the relationship between the composition, dissolution rate, ion release and cytocompatibility of a series of borophosphate glasses. While, the base glass was selected to be 40mol%P2O5-16mol%CaO-24mol%MgO-20mol%Na2O, three B2O3 modified glass compositions were formulated by replacing Na2O with 1, 5 and 10 mol% B2O3. Ion release study was conducted using inductively coupled plasma atomic emission spectroscopy (ICP-AES). The thermal scans of the glasses as determined by differential scanning calorimetry (DSC) revealed an increment in the thermal properties with increasing B2O3 content in the glasses. On the other hand, the dissolution rate of the glasses decreased with increasing B2O3 content. To identify the effect of boron ion release on the cytocompatibility properties of the glasses, MG63 cells were cultured on the surface of the glass discs. The in vitro cell culture study suggested that glasses with 5 mol% B2O3 (P40B5) showed better cell proliferation and metabolic activity as compares to the glasses with 10 mol% (P40B10) or with no B2O3 (P40B0). The confocal laser scanning microscopy (CLSM) images of live/dead stained MG63 cells attached to the surface of the glasses also revealed that the number of dead cells attached to P40B5 glasses were significantly lower than both P40B0 and P40B10 glasses.

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