A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy
Objectives: To test and validate a new protocol for in vitro contamination of dentinal tubules using Fusobacterium nucleatum (F. nucleatum) by confocal laser scanning microscopy (CLSM), in addition to evaluating the effectiveness of conventional endodontic irrigants such as sodium hypochlorite (NaOC...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-07-01
|
| Series: | Heliyon |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844023052507 |
| _version_ | 1827892292413816832 |
|---|---|
| author | Mirela Cesar Barros Victor Feliz Pedrinha Marcia Sirlene Zardin Graeff Clovis Monteiro Bramante Marco Antonio Hungaro Duarte Flaviana Bombarda de Andrade |
| author_facet | Mirela Cesar Barros Victor Feliz Pedrinha Marcia Sirlene Zardin Graeff Clovis Monteiro Bramante Marco Antonio Hungaro Duarte Flaviana Bombarda de Andrade |
| author_sort | Mirela Cesar Barros |
| collection | DOAJ |
| description | Objectives: To test and validate a new protocol for in vitro contamination of dentinal tubules using Fusobacterium nucleatum (F. nucleatum) by confocal laser scanning microscopy (CLSM), in addition to evaluating the effectiveness of conventional endodontic irrigants such as sodium hypochlorite (NaOCl) and chlorhexidine (CLX) on this biofilm. Material and methods: Thirty lower premolars were contaminated with F. nucleatum (ATCC 51190) for 7 days under anaerobic conditions using the proposed new model. The specimens were divided into a control group and experimental groups, according to the irrigants: NaOCl 2.5% and CLX 2%. Then, the samples were submitted for analysis by CLSM and the LIVE/DEAD technique to quantify bacterial viability. Data normality was verified by the Shapiro-Wilk test. Intragroup and intergroup comparisons were performed using the Kruskal-Wallis test, followed by Dunn's post-test. Results: The CLSM images obtained demonstrated the effectiveness of the proposed new contamination protocol, with a high percentage of viable bacteria in relation to the treated groups (p < 0.05). Lower viability values were observed for the 2.5% NaOCl group. Conclusion: The new contamination protocol resulted in a high and homogeneous percentage of viable bacteria in the dentinal tubules in all specimens evaluated. Both irrigating solutions proved to be effective in reducing the intratubular microbiota, especially 2.5% NaOCl. |
| first_indexed | 2024-03-12T21:37:41Z |
| format | Article |
| id | doaj.art-a4a6bbfafcc54abca4bcb4a9f27b9e36 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-03-12T21:37:41Z |
| publishDate | 2023-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-a4a6bbfafcc54abca4bcb4a9f27b9e362023-07-27T05:58:20ZengElsevierHeliyon2405-84402023-07-0197e18042A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopyMirela Cesar Barros0Victor Feliz Pedrinha1Marcia Sirlene Zardin Graeff2Clovis Monteiro Bramante3Marco Antonio Hungaro Duarte4Flaviana Bombarda de Andrade5Department of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru, SP, BrazilDepartment of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru, SP, BrazilIntegrated Research Center, Bauru School of Dentistry, University of São Paulo, Bauru, SP, BrazilDepartment of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru, SP, BrazilDepartment of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru, SP, BrazilDepartment of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil; Corresponding author. Bauru School of Dentistry, University of São Paulo, Al. Dr. Octávio Pinheiro Brisolla 9-75, Vila Universitária, 17012-901, Bauru, SP, Brazil.Objectives: To test and validate a new protocol for in vitro contamination of dentinal tubules using Fusobacterium nucleatum (F. nucleatum) by confocal laser scanning microscopy (CLSM), in addition to evaluating the effectiveness of conventional endodontic irrigants such as sodium hypochlorite (NaOCl) and chlorhexidine (CLX) on this biofilm. Material and methods: Thirty lower premolars were contaminated with F. nucleatum (ATCC 51190) for 7 days under anaerobic conditions using the proposed new model. The specimens were divided into a control group and experimental groups, according to the irrigants: NaOCl 2.5% and CLX 2%. Then, the samples were submitted for analysis by CLSM and the LIVE/DEAD technique to quantify bacterial viability. Data normality was verified by the Shapiro-Wilk test. Intragroup and intergroup comparisons were performed using the Kruskal-Wallis test, followed by Dunn's post-test. Results: The CLSM images obtained demonstrated the effectiveness of the proposed new contamination protocol, with a high percentage of viable bacteria in relation to the treated groups (p < 0.05). Lower viability values were observed for the 2.5% NaOCl group. Conclusion: The new contamination protocol resulted in a high and homogeneous percentage of viable bacteria in the dentinal tubules in all specimens evaluated. Both irrigating solutions proved to be effective in reducing the intratubular microbiota, especially 2.5% NaOCl.http://www.sciencedirect.com/science/article/pii/S2405844023052507Fusobacterium nucleatumConfocal microscopyContaminationDentin |
| spellingShingle | Mirela Cesar Barros Victor Feliz Pedrinha Marcia Sirlene Zardin Graeff Clovis Monteiro Bramante Marco Antonio Hungaro Duarte Flaviana Bombarda de Andrade A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy Heliyon Fusobacterium nucleatum Confocal microscopy Contamination Dentin |
| title | A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy |
| title_full | A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy |
| title_fullStr | A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy |
| title_full_unstemmed | A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy |
| title_short | A new model of in vitro dentin intratubular contamination for Fusobacterium nucleatum: Validation by confocal laser scanning microscopy |
| title_sort | new model of in vitro dentin intratubular contamination for fusobacterium nucleatum validation by confocal laser scanning microscopy |
| topic | Fusobacterium nucleatum Confocal microscopy Contamination Dentin |
| url | http://www.sciencedirect.com/science/article/pii/S2405844023052507 |
| work_keys_str_mv | AT mirelacesarbarros anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT victorfelizpedrinha anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT marciasirlenezardingraeff anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT clovismonteirobramante anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT marcoantoniohungaroduarte anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT flavianabombardadeandrade anewmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT mirelacesarbarros newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT victorfelizpedrinha newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT marciasirlenezardingraeff newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT clovismonteirobramante newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT marcoantoniohungaroduarte newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy AT flavianabombardadeandrade newmodelofinvitrodentinintratubularcontaminationforfusobacteriumnucleatumvalidationbyconfocallaserscanningmicroscopy |