ABSTRACT
Periodontal diseases manifest by the formation of deep pockets between the gingiva and teeth where multispecies bacterial biofilms flourish, causing inflammation and bone loss. Epithelial cell receptor αvß6 integrin that regulates inflammation by activating the anti-inflammatory cytokine transforming growth factor-ß1, is highly expressed in healthy junctional epithelium that connects the gingiva to the tooth enamel. However, its expression is attenuated in human periodontal disease. Moreover, Itgb6 -/- mice display increased periodontal inflammation compared to wild-type mice. We hypothesized that bacterial biofilms present in the periodontal pockets suppress αvß6 integrin levels in periodontal disease and that this change aggravates inflammation. To this end, we generated three-week-old multi-species oral biofilms in vitro and treated cultured gingival epithelial cells (GECs) with their extracts. The biofilm extracts caused suppression of ß6 integrin expression and upregulation of pro-inflammatory cytokines, including interleukin-1ß and -6. Furthermore, GECs with ß6 integrin siRNA knockdown showed increased interleukin-1ß expression, indicating that αvß6 integrin-deficiency is associated with pro-inflammatory cytokine responsiveness. FSL-1, a synthetic bacterial lipopeptide, also suppressed ß6 integrin expression in GECs. Therefore, biofilm components, including lipopeptides, may downregulate αvß6 integrin expression in the pocket epithelium and thus promote epithelial cell-driven pro-inflammatory response in periodontal disease.
Subject(s)
Antigens, Neoplasm/genetics , Biofilms , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Gingiva/cytology , Gingiva/microbiology , Integrins/genetics , Microbiota , Animals , Cytokines/metabolism , Dental Plaque/microbiology , Diglycerides/metabolism , Gene Expression , Gene Knockout Techniques , Humans , Inflammation Mediators/metabolism , Keratinocytes/metabolism , Mice , Oligopeptides/metabolism , Periodontal Diseases/genetics , Periodontal Diseases/metabolism , Periodontal Diseases/pathology , Signal Transduction , Transforming Growth Factor beta1/metabolismABSTRACT
Calcium hydroxide (CH) dressing residues can compromise endodontic sealing. This study aimed to evaluate the amount of remaining CH in root canals after mechanical removal by four groups of irrigation techniques including needle irrigation only, ProTaper file, EndoActivator, and ultrasonic file. Fifteen extracted single-rooted teeth were collected and used for all four groups. The samples were firstly prepared by ProTaper rotary instruments, and then sectioned longitudinally through the long axis of the root canals, followed by final reassembling by wires. CH was kept in the canals for 7 days setting. The removal procedure began with 5 mL of 2.5% sodium hypochlorite (NaOCl) followed by 1 mL of 17% ethylenediaminetetraacetic acid and a final irrigation with 5 mL of 2.5% NaOCl solution for all groups. No additional agitation of the irrigant was performed in group 1, while agitation for 20 s between irrigants was done with F2 ProTaper rotary file in group 2, EndoActivator with tip size 25/.04 in group 3 and by an ultrasonic file 25/.02 in group 4. The total activation time was 60 s. The roots were then disassembled and captured by digital camera. The ratio of CH coated surface area to the surface area of the whole canal as well as each third of the canal was calculated. The data were statistically analyzed by one-way ANOVA using post hoc Tukey test. Results showed that none of the four techniques could remove all CH. No significant difference was found between EndoActivator and ultrasonic techniques. However, they both removed significantly more CH than ProTaper and needle irrigation (P=0.0001). In conclusion, the sonic and ultrasonic agitation techniques were more effective in removing intracanal medicaments than the ProTaper rotary file and needle irrigation in all thirds of the canal.
