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.

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 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.

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 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 ROOT CEMENTUM INVASION AND ADHESION BY ENTEROCOCCUS FAECALIS CONFOCAL ANALYSIS.

2012 ◽  
Vol 02 (04) ◽  
pp. 44-49
Author(s):  
Rahul Halkai ◽  
Mithra N. Hegde ◽  
Kiran Halkai
Keyword(s):  
Enterococcus Faecalis ◽  
Root Canal ◽  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Control Group ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Group I ◽  
Group Ii

Abstract Aims to know ability of Enterococcus faecalis invasion and adhesion into root cementum. Methodology Forty single rooted human intact teeth were selected, group I with 20 teeth as control group with no access opening and apical seal with varnish. Group II with 20 teeth, after access opening and canal debridement, all the samples were subjected for gamma sterilization to ensure complete absence of microorganisms, and then exposed to Enterococcus faecalis broth, in group II broth is placed with the help of micro pipette into root canal and at the same time apical 1/3 of tooth were immersed into broth with both groups for 8 weeks. Biomechanical preparation, obturation and coronal sealing done using GIC with group II, followed by apical 1/3 immersed in the enterococcus broth for 8 weeks, examination under confocal laser scanning microscope after splitting the teeth samples into two halfs buccolingually. Results: This study shows only adhesion of Enterococcus faecalis to root cementum. Conclusion Adhesion of Enterococcus faecalis to root cementum may be the possible reason for persisting infection after root canal treatment.

scholarly journals The Penetration Ability of Calcium Silicate Root Canal Sealers into Dentinal Tubules Compared to Conventional Resin-Based Sealer: A Confocal Laser Scanning Microscopy Study

Materials ◽  
2019 ◽  
Vol 12 (3) ◽  
pp. 531 ◽  
Author(s):  
Yemi Kim ◽  
Ban-Suk Kim ◽  
Yong-Min Kim ◽  
Donghee Lee ◽  
Sin-Young Kim
Keyword(s):  
Root Canal ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Intensity Level ◽  
Single Cone ◽  
Penetration Ability ◽  
Ah Plus ◽  
Root Canal Sealers ◽  
Apical Area

The purpose of this study was to compare the penetration ability of calcium silicate root canal sealers and conventional resin-based sealer using confocal laser scanning microscopy (CLSM). A total of 60 recently extracted single-rooted human premolars were used in this study. The root canals were prepared to a size 40/0.06 taper with ProFile rotary instruments and irrigated with NaOCl and EDTA. After drying all canals, the specimens were randomly divided into three experimental groups (n = 20): Group 1, gutta-percha (GP)/AH Plus with continuous wave compaction; group 2, GP/BioRoot RCS with a single-cone technique; and group 3, GP/Endoseal MTA with a single-cone technique. All experimental samples were sectioned perpendicular to their long axis using a low-speed diamond wheel at the apical, middle, and coronal third levels. The penetration abilities of all samples were evaluated using CLSM. A Kruskal–Wallis analysis and a series of Mann–Whitney U post hoc tests were performed. A higher intensity level was found in the coronal area and a lower intensity level in the apical area in all the experimental groups. The AH Plus group showed higher sum fluorescence intensity in the apical and coronal thirds compared with the BioRoot RCS and Endoseal MTA groups, whereas the BioRoot RCS group showed a higher intensity level in the middle third, similar to the AH Plus group. The maximum sealer penetration depth was low in the apical area and high in the coronal area in the AH Plus and Endoseal MTA groups. In the BioRoot RCS group, maximum sealer penetration was observed in the middle third. In conclusion, there were significant differences in sealer penetration pattern and distance according to the root level and sealer type.

scholarly journals Evaluation of four final irrigation protocols for cleaning root canal walls

2020 ◽  
Vol 12 (1) ◽  
Author(s):  
Qiang Li ◽  
Qian Zhang ◽  
Xiaoying Zou ◽  
Lin Yue
Keyword(s):  
Root Canal ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Smear Layer ◽  
Confocal Laser ◽  
Bacterial Survival ◽  
Bacterial Inhibition ◽  
Layer Removal ◽  
Significant Difference ◽  
Canal Systems

Abstract The aim of this study was to compare the efficiency of four final irrigation protocols in smear layer removal and bacterial inhibition in root canal systems. Thirty roots inoculated with Enterococcus faecalis were prepared with ProTaper Universal files. The teeth were disinfected by conventional needle irrigation, sonic agitation using the EndoActivator device, passive ultrasonic irrigation, or an M3 Max file. Teeth with no root canal preparation served as blank controls for the establishment of the infection baseline. Teeth with preparation but no final irrigation served as a post-instrumentation baseline. After the final irrigation, the teeth were sectioned in half. One half of each tooth was examined by scanning electron microscopy (SEM) to assess smear layer removal using a five-point scale. The other half was examined by confocal laser scanning microscopy (CLSM) using the LIVE/DEAD BackLight bacterial viability kit to evaluate the depth of bacterial survival in dentinal tubules. SEM analysis revealed no significant difference in smear layer removal throughout the whole canal among the EA, PUI, and M3 Max groups (P > 0.05). CLSM revealed that PUI achieved the greatest bacterial inhibition depth in the coronal ((174.27 ± 31.63) μm), middle ((160.94 ± 37.77) μm), and apical ((119.53 ± 28.49) μm) thirds of the canal (all P < 0.05 vs. other groups). According to this comprehensive SEM and CLSM evaluation, PUI appears to have the best infection control ability in root canal systems.

scholarly journals Comparative Evaluation of Apical Sealing Ability of Different Obturation Techniques by Confocal Microscopy: An In Vitro Study

2020 ◽  
Vol 8 (02) ◽  
pp. 55-59
Author(s):  
Reetu Arora ◽  
Yogesh Kumar ◽  
Neetu Jindal ◽  
Renu Aggarwal ◽  
Kavneet Takhar
Keyword(s):  
Root Canal ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Group Iv ◽  
Control Group ◽  
Group Vi ◽  
Group V ◽  
Group Iii ◽  
Group I

Abstract Introduction The aim of obturation in the root canal is to completely seal the canal space to eliminate all the portals of entry and exit between root canal and periodontal space. Various techniques have been developed to achieve a hermetic seal. Materials and Methods As many as 150 extracted human maxillary central incisors were taken for the study. Biomechanical preparation was done up to F5 protaper file. According to different obturation techniques, samples were divided into six groups, keeping 30 samples in experimental and 15 samples in control groups. Group I–Lateral Condensation, Group II–Thermafil, Group III–Beefill, Group IV–GuttaFlow, Group V–Positive Control group, Group VI–Negative Control group. After obturation, the samples were immersed in 2% Rhodamine-B dye for 24 hours. Each sample was longitudinally sectioned to examine under confocal laser scanning microscope. Statistical Analysis The results were evaluated with ANOVA and posthoc Tukey honest significant difference (HSD) comparison test. Results The mean values of dye penetration of different groups were Group I (Lateral Condensation) 1.51 ± 0.451, Group II (Thermafil) 0.918 ± 0.399, Group III (Beefill) 1.30 ± 0.559. Group IV (GuttaFlow) 0.655 ± 0.396, Group V (Positive Control group) 1.96 ±0.046, Group VI (Negative Control group) 0 ± 0. The lowest mean value of apical microleakage was found in GuttaFlow amongst all experimental groups. Conclusion It can be concluded that the GuttaFlow obturating material exhibited better apical sealing ability with canal walls.

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