Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite
Biofilm communities are tolerant to antimicrobials and difficult to eradicate. This study aimed to investigate the effect of melittin, an antimicrobial peptide, either alone or in combination with deoxyribonuclease (DNase), an inhibitor of extracellular deoxyribonucleic acid (eDNA), against <i>...
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doaj-32ced153adf04f13850f67ceb54ea0dd2020-11-25T03:02:11ZengMDPI AGMaterials1996-19442020-08-01133740374010.3390/ma13173740Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium HypochloriteSujitha Ramaraj0Mi-Ah Kim1Vinicius Rosa2Prasanna Neelakantan3Won-Jun Shon4Kyung-San Min5Department of Conservative Dentistry, School of Dentistry and Institute of Oral Bioscience, Jeonbuk National University, Jeonju 54896, KoreaDepartment of Conservative Dentistry, School of Dentistry and Institute of Oral Bioscience, Jeonbuk National University, Jeonju 54896, KoreaDiscipline of Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore 119085, SingaporeDiscipline of Endodontology, Department of Restorative Dental Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, ChinaDepartment of Conservative Dentistry, School of Dentistry, Seoul National University, Seoul 03080, KoreaDepartment of Conservative Dentistry, School of Dentistry and Institute of Oral Bioscience, Jeonbuk National University, Jeonju 54896, KoreaBiofilm communities are tolerant to antimicrobials and difficult to eradicate. This study aimed to investigate the effect of melittin, an antimicrobial peptide, either alone or in combination with deoxyribonuclease (DNase), an inhibitor of extracellular deoxyribonucleic acid (eDNA), against <i>Enterococcus faecalis</i> (<i>E. faecalis</i>) biofilms, and biofilm susceptibility to sodium hypochlorite (NaOCl). Biofilms of <i>E. faecalis</i> were developed in root canals of bovine teeth. The biofilms were treated with distilled water (control), melittin, DNase, or DNase+melittin. The antibiofilm effects of the treatments were analyzed using colony forming unit (CFU) assay, crystal violet staining, confocal laser scanning microscopy (CLSM), and field emission scanning electron microscope (FE-SEM). The susceptibility of DNase+melittin-treated biofilms to NaOCl (0%, 2.5% and 5%) was investigated by the CFU assay. The data were statistically analyzed using one-way analysis of variance, followed by Tukey’s test. A p-value of <0.05 was considered significant. Specimens treated with DNase+melittin showed a more significant decrease in the CFUs, eDNA level, and biofilm formation rate than those treated only with melittin or DNase (<i>p</i> < 0.05). CLSM analysis showed DNase+melittin treatment significantly reduced the volume of biofilms and extracellular polymeric substance compared to either treatment alone (<i>p</i> < 0.05). FE-SEM images showed a high degree of biofilm disruption in specimens that received DNase+melittin. 2.5% NaOCl in specimens pretreated with DNase+melittin showed higher antibacterial activity than those treated only with 5% NaOCl (<i>p</i> < 0.05). This study highlighted that DNase improved the antibiofilm effects of melittin. Moreover, DNase+melittin treatment increased the susceptibility of biofilms to NaOCl. Thus, the complex could be a clinical strategy for safer use of NaOCl by reducing the concentration.https://www.mdpi.com/1996-1944/13/17/3740biofilmDNase<i>Enterococcus faecalis</i>melittinsodium hypochlorite |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sujitha Ramaraj Mi-Ah Kim Vinicius Rosa Prasanna Neelakantan Won-Jun Shon Kyung-San Min |
spellingShingle |
Sujitha Ramaraj Mi-Ah Kim Vinicius Rosa Prasanna Neelakantan Won-Jun Shon Kyung-San Min Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite Materials biofilm DNase <i>Enterococcus faecalis</i> melittin sodium hypochlorite |
author_facet |
Sujitha Ramaraj Mi-Ah Kim Vinicius Rosa Prasanna Neelakantan Won-Jun Shon Kyung-San Min |
author_sort |
Sujitha Ramaraj |
title |
Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite |
title_short |
Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite |
title_full |
Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite |
title_fullStr |
Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite |
title_full_unstemmed |
Combined Effect of Melittin<i> </i>and DNase on <i>Enterococcus</i> f<i>aecalis</i> Biofilms and Its Susceptibility to Sodium Hypochlorite |
title_sort |
combined effect of melittin<i> </i>and dnase on <i>enterococcus</i> f<i>aecalis</i> biofilms and its susceptibility to sodium hypochlorite |
publisher |
MDPI AG |
series |
Materials |
issn |
1996-1944 |
publishDate |
2020-08-01 |
description |
Biofilm communities are tolerant to antimicrobials and difficult to eradicate. This study aimed to investigate the effect of melittin, an antimicrobial peptide, either alone or in combination with deoxyribonuclease (DNase), an inhibitor of extracellular deoxyribonucleic acid (eDNA), against <i>Enterococcus faecalis</i> (<i>E. faecalis</i>) biofilms, and biofilm susceptibility to sodium hypochlorite (NaOCl). Biofilms of <i>E. faecalis</i> were developed in root canals of bovine teeth. The biofilms were treated with distilled water (control), melittin, DNase, or DNase+melittin. The antibiofilm effects of the treatments were analyzed using colony forming unit (CFU) assay, crystal violet staining, confocal laser scanning microscopy (CLSM), and field emission scanning electron microscope (FE-SEM). The susceptibility of DNase+melittin-treated biofilms to NaOCl (0%, 2.5% and 5%) was investigated by the CFU assay. The data were statistically analyzed using one-way analysis of variance, followed by Tukey’s test. A p-value of <0.05 was considered significant. Specimens treated with DNase+melittin showed a more significant decrease in the CFUs, eDNA level, and biofilm formation rate than those treated only with melittin or DNase (<i>p</i> < 0.05). CLSM analysis showed DNase+melittin treatment significantly reduced the volume of biofilms and extracellular polymeric substance compared to either treatment alone (<i>p</i> < 0.05). FE-SEM images showed a high degree of biofilm disruption in specimens that received DNase+melittin. 2.5% NaOCl in specimens pretreated with DNase+melittin showed higher antibacterial activity than those treated only with 5% NaOCl (<i>p</i> < 0.05). This study highlighted that DNase improved the antibiofilm effects of melittin. Moreover, DNase+melittin treatment increased the susceptibility of biofilms to NaOCl. Thus, the complex could be a clinical strategy for safer use of NaOCl by reducing the concentration. |
topic |
biofilm DNase <i>Enterococcus faecalis</i> melittin sodium hypochlorite |
url |
https://www.mdpi.com/1996-1944/13/17/3740 |
work_keys_str_mv |
AT sujitharamaraj combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite AT miahkim combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite AT viniciusrosa combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite AT prasannaneelakantan combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite AT wonjunshon combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite AT kyungsanmin combinedeffectofmelittiniianddnaseonienterococcusifiaecalisibiofilmsanditssusceptibilitytosodiumhypochlorite |
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1724690950177947648 |