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

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 enlar...

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Autores principales: Parolia, Abhishek, Kumar, Haresh, Ramamurthy, Srinivasan, Madheswaran, Thiagarajan, Davamani, Fabian, Pichika, Malikarjuna Rao, Mak, Kit-Kay, Fawzy, Amr S, Daood, Umer, Pau, Allan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7866495/
https://www.ncbi.nlm.nih.gov/pubmed/33573147
http://dx.doi.org/10.3390/molecules26030715
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author Parolia, Abhishek
Kumar, Haresh
Ramamurthy, Srinivasan
Madheswaran, Thiagarajan
Davamani, Fabian
Pichika, Malikarjuna Rao
Mak, Kit-Kay
Fawzy, Amr S
Daood, Umer
Pau, Allan
author_facet Parolia, Abhishek
Kumar, Haresh
Ramamurthy, Srinivasan
Madheswaran, Thiagarajan
Davamani, Fabian
Pichika, Malikarjuna Rao
Mak, Kit-Kay
Fawzy, Amr S
Daood, Umer
Pau, Allan
author_sort Parolia, Abhishek
collection PubMed
description 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.
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spelling pubmed-78664952021-02-07 Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal Parolia, Abhishek Kumar, Haresh Ramamurthy, Srinivasan Madheswaran, Thiagarajan Davamani, Fabian Pichika, Malikarjuna Rao Mak, Kit-Kay Fawzy, Amr S Daood, Umer Pau, Allan Molecules Article 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. MDPI 2021-01-30 /pmc/articles/PMC7866495/ /pubmed/33573147 http://dx.doi.org/10.3390/molecules26030715 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Parolia, Abhishek
Kumar, Haresh
Ramamurthy, Srinivasan
Madheswaran, Thiagarajan
Davamani, Fabian
Pichika, Malikarjuna Rao
Mak, Kit-Kay
Fawzy, Amr S
Daood, Umer
Pau, Allan
Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title_full Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title_fullStr Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title_full_unstemmed Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title_short Effect of Propolis Nanoparticles against Enterococcus faecalis Biofilm in the Root Canal
title_sort effect of propolis nanoparticles against enterococcus faecalis biofilm in the root canal
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7866495/
https://www.ncbi.nlm.nih.gov/pubmed/33573147
http://dx.doi.org/10.3390/molecules26030715
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