Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm

Fabíola Galbiatti de Carvalho, Regina Maria Puppin-Rontani, Suzana Beatriz Portugal de Fúcio, Thais de Cássia Negrini, Hugo Lemes Carlo, Franklin Garcia-Godoy

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Since bacteria remain in the dentin following caries removal, restorative materials with antibacterial properties are desirable to help maintaining the residual microorganisms inactive. The adhesive system Clearfil Protect Bond (PB) contains the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) in its primer, which has shown antimicrobial activity. However, its bactericidal effect against biofilm on the dentin has been little investigated. Objective: The aim of this study was to analyze by confocal laser scanning microscopy (CLSM) and viable bacteria counting (CFU) the MDPB bactericidal effect against S. mutans biofilm on the dentin surface. Material and Methods: Bovine dentin surfaces were obtained and subjected to S. mutans biofilm formation in BHI broth supplemented with 1% (w/v) sucrose for 18 h. Samples were divided into three groups, according to the primer application (n=3): Clearfil Protect Bond (PB), Clearfil SE Bond, which does not contain MDPB, (SE) and saline (control group). After the biofilm formation, Live/ Dead stain was applied directly to the surface of each sample. Next, 10 μL of each primer were applied on the samples during 590 s for the real-time CLSM analysis. The experiment was conducted in triplicate. The primers and saline were also applied on the other dentin samples during 20, 90, 300 and 590 s (n=9 for each group and period evaluated) and the CFU were assessed by colonies counting. Results: The results of the CLSM showed that with the SE application, although non-viable bacteria were detected at 20 s, there was no increase in their count during 590 s. In contrast, after the PB application there was a gradual increase of non-viable bacteria over 590 s. Conclusions: The quantitative analysis demonstrated a significant decrease of S. mutans CFU at 90 s PB exposure and only after 300 s of SE application. Protect Bond showed an earlier antibacterial effect than SE Bond.

Original languageEnglish (US)
Pages (from-to)568-575
Number of pages8
JournalJournal of Applied Oral Science
Volume20
Issue number5
DOIs
StatePublished - Jan 1 2012

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Dentin
Biofilms
Confocal Microscopy
Bacteria
Adhesives
Sucrose
Coloring Agents
methacryloyloxydodecylpyridinium bromide
Clearfil Protect Bond
Control Groups

All Science Journal Classification (ASJC) codes

  • Dentistry(all)

Cite this

Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm. / de Carvalho, Fabíola Galbiatti; Puppin-Rontani, Regina Maria; de Fúcio, Suzana Beatriz Portugal; de Cássia Negrini, Thais; Carlo, Hugo Lemes; Garcia-Godoy, Franklin.

In: Journal of Applied Oral Science, Vol. 20, No. 5, 01.01.2012, p. 568-575.

Research output: Contribution to journalArticle

de Carvalho, Fabíola Galbiatti ; Puppin-Rontani, Regina Maria ; de Fúcio, Suzana Beatriz Portugal ; de Cássia Negrini, Thais ; Carlo, Hugo Lemes ; Garcia-Godoy, Franklin. / Analysis by confocal laser scanning microscopy of the MDPB bactericidal effect on S. mutans biofilm CLSM analysis of MDPB bactericidal effect on biofilm. In: Journal of Applied Oral Science. 2012 ; Vol. 20, No. 5. pp. 568-575.
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abstract = "Since bacteria remain in the dentin following caries removal, restorative materials with antibacterial properties are desirable to help maintaining the residual microorganisms inactive. The adhesive system Clearfil Protect Bond (PB) contains the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) in its primer, which has shown antimicrobial activity. However, its bactericidal effect against biofilm on the dentin has been little investigated. Objective: The aim of this study was to analyze by confocal laser scanning microscopy (CLSM) and viable bacteria counting (CFU) the MDPB bactericidal effect against S. mutans biofilm on the dentin surface. Material and Methods: Bovine dentin surfaces were obtained and subjected to S. mutans biofilm formation in BHI broth supplemented with 1{\%} (w/v) sucrose for 18 h. Samples were divided into three groups, according to the primer application (n=3): Clearfil Protect Bond (PB), Clearfil SE Bond, which does not contain MDPB, (SE) and saline (control group). After the biofilm formation, Live/ Dead stain was applied directly to the surface of each sample. Next, 10 μL of each primer were applied on the samples during 590 s for the real-time CLSM analysis. The experiment was conducted in triplicate. The primers and saline were also applied on the other dentin samples during 20, 90, 300 and 590 s (n=9 for each group and period evaluated) and the CFU were assessed by colonies counting. Results: The results of the CLSM showed that with the SE application, although non-viable bacteria were detected at 20 s, there was no increase in their count during 590 s. In contrast, after the PB application there was a gradual increase of non-viable bacteria over 590 s. Conclusions: The quantitative analysis demonstrated a significant decrease of S. mutans CFU at 90 s PB exposure and only after 300 s of SE application. Protect Bond showed an earlier antibacterial effect than SE Bond.",
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