FISHing for gutta-percha-adhered biofilms in purulent post-treatment apical periodontitis.
This study investigated the possibility of depicting individual taxa in clinically relevant biofilms using fluorescent in situ hybridization (FISH). Gutta-percha samples were collected from the apical aspect of root canals associated with a chronic apical abscess (test samples, n = 8). Corresponding control samples were obtained from previously filled root canals with apparently normal periapical tissues (n = 3). The transport medium was investigated for detached biofilm fragments using FISH staining and conventional epifluorescence microscopy. Gutta-percha samples were stained by multiplex FISH, and inspected using confocal laser scanning microscopy. FISH of the transport medium confirmed the presence of the main species formerly identified by conventional methods in post-treatment purulent endodontic infections, most prominently Fusobacterium spp., Bacteroidetes and Prevotellaceae. Treponemes were identified in five of eight cases associated with purulent infections, but Enterococcus faecalis and Staphylococcus spp. were not identified. The biofilms on gutta-percha from root canals associated with apical periodontitis showed dense aggregates of variable composition. Control samples contained few, if any, bacteria in the transport medium, and featured no biofilms on the respective gutta-percha specimens. The current study revealed some direct, visual in situ information on the nature of biofilms associated with purulent periapical infections in man.
Clinic of Preventive Dentistry, Periodontology and Cariology, Center for Dental Medicine, University of Zürich, Zürich, Switzerland.
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M
Zehnder
Clinic of Preventive Dentistry, Periodontology and Cariology, Center for Dental Medicine, University of Zürich, Zürich, Switzerland.
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DK
Rechenberg
Division of Oral Microbiology and Immunology, Institute of Oral Biology, Center for Dental Medicine, University of Zürich, Zürich, Switzerland.
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T
Thurnheer
Division of Oral Microbiology and Immunology, Institute of Oral Biology, Center for Dental Medicine, University of Zürich, Zürich, Switzerland.
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H
Lüthi-Schaller
Division of Oral Microbiology and Immunology, Institute of Oral Biology, Center for Dental Medicine, University of Zürich, Zürich, Switzerland.|Division of Cariology and Endodontics, Department of Dental Medicine, Karolinska Institute, Huddinge, Sweden.
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Belibasakis
D000328:Adult; D000368:Aged; D041963:Bacteroidetes / Q000302:isolation & purification; D018441:Biofilms; D003786:Dental Pulp Cavity / Q000382:microbiology; D013293:Enterococcus faecalis / Q000302:isolation & purification; D005260:Female; D005673:Fusobacterium / Q000302:isolation & purification; D006171:Gutta-Percha / Q000737:chemistry*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010485:Periapical Periodontitis / Q000382:microbiology* / Q000628:therapy; D018720:Prevotella / Q000302:isolation & purification; D013210:Staphylococcus / Q000302:isolation & purification
D006171:Gutta-Percha
apical periodontitis; biofilms; fluorescence in situ hybridization; infection; root canal