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A new improved protocol for in vitro intratubular dentinal bacterial contamination for antimicrobial endodontic tests: standardization and validation by confocal laser scanning microscopy

Overview of attention for article published in Journal of Applied Oral Science, July 2015
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Title
A new improved protocol for in vitro intratubular dentinal bacterial contamination for antimicrobial endodontic tests: standardization and validation by confocal laser scanning microscopy
Published in
Journal of Applied Oral Science, July 2015
DOI 10.1590/1678-775720140261
Pubmed ID
Authors

ANDRADE, Flaviana Bombarda de, ARIAS, Marcela Paola Castro, MALIZA, Amanda Garcia Alves, DUARTE, Marco Antonio Hungaro, GRAEFF, Márcia Sirlene Zardin, AMOROSO-SILVA, Pablo Andrés, MIDENA, Raquel Zanin, MORAES, Ivaldo Gomes de, Andrade, Flaviana Bombarda de, Arias, Marcela Paola Castro, Maliza, Amanda Garcia Alves, Duarte, Marco Antonio Hungaro, Graeff, Márcia Sirlene Zardin, Amoroso-Silva, Pablo Andrés, Midena, Raquel Zanin, Moraes, Ivaldo Gomes de

Abstract

Objectives To compare three methods of intratubular contamination that simulate endodontic infections using confocal laser scanning microscopy (CLSM). Material and Methods Two pre-existing models of dentinal contamination were used to induce intratubular infection (groups A and B). These methods were modified in an attempt to improve the model (group C). Among the modifications it may be included: specimen contamination for five days, ultrasonic bath with BHI broth after specimen sterilization, use of E. faecalis during the exponential growth phase, greater concentration of inoculum, and two cycles of centrifugation on alternate days with changes of culture media. All specimens were longitudinally sectioned and stained with of LIVE/DEAD® for 20 min. Specimens were assessed using CLSM, which provided images of the depth of viable bacterial proliferation inside the dentinal tubules. Additionally, three examiners used scores to classify the CLSM images according to the following parameters: homogeneity, density, and depth of the bacterial contamination inside the dentinal tubules. Kruskal-Wallis and Dunn's tests were used to evaluate the live and dead cells rates, and the scores obtained. Results The contamination scores revealed higher contamination levels in group C when compared with groups A and B (p<0.05). No differences were observed between group A and B (p>0.05). The volume of live cells in group C was higher than in groups A and B (p<0.05). Conclusion The new protocol for intratubular infection resulted in high and uniform patterns of bacterial contamination and higher cell viability in all specimens when compared with the current methods.

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Mendeley readers

The data shown below were compiled from readership statistics for 57 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Egypt 1 2%
Unknown 56 98%

Demographic breakdown

Readers by professional status Count As %
Student > Master 12 21%
Student > Ph. D. Student 8 14%
Student > Bachelor 6 11%
Student > Doctoral Student 5 9%
Student > Postgraduate 5 9%
Other 9 16%
Unknown 12 21%
Readers by discipline Count As %
Medicine and Dentistry 34 60%
Agricultural and Biological Sciences 3 5%
Biochemistry, Genetics and Molecular Biology 2 4%
Nursing and Health Professions 1 2%
Immunology and Microbiology 1 2%
Other 1 2%
Unknown 15 26%