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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
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; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • ARIAS, Marcela Paola Castro; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • MALIZA, Amanda Garcia Alves; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • DUARTE, Marco Antonio Hungaro; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • GRAEFF, Márcia Sirlene Zardin; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • AMOROSO-SILVA, Pablo Andrés; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • MIDENA, Raquel Zanin; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
  • MORAES, Ivaldo Gomes de; University of São Paulo. Bauru School of Dentistry,. Department of Operative Dentistry, Endodontics and Dental Materials. Bauru. BR
J. appl. oral sci ; 23(6): 591-598, Nov.-Dec. 2015. graf
Article in English | LILACS, BBO | ID: lil-769816
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. faecalisduring 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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Full text: Available Index: LILACS (Americas) Main subject: Enterococcus faecalis / Dental Pulp Cavity / Dentin / Disease Models, Animal Type of study: Prognostic study Limits: Animals Language: English Journal: J. appl. oral sci Journal subject: Dentistry Year: 2015 Type: Article Affiliation country: Brazil Institution/Affiliation country: University of São Paulo/BR

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Full text: Available Index: LILACS (Americas) Main subject: Enterococcus faecalis / Dental Pulp Cavity / Dentin / Disease Models, Animal Type of study: Prognostic study Limits: Animals Language: English Journal: J. appl. oral sci Journal subject: Dentistry Year: 2015 Type: Article Affiliation country: Brazil Institution/Affiliation country: University of São Paulo/BR