Truncating NFKB1 variants cause combined NLRP3 inflammasome activation and type I interferon signaling and predispose to necrotizing fasciitis.

In monogenic autoinflammatory diseases, mutations in genes regulating innate immune responses often lead to uncontrolled activation of inflammasome pathways or the type I interferon (IFN-I) response. We describe a mechanism of autoinflammation potentially predisposing patients to life-threatening necrotizing soft tissue inflammation. Six unrelated families are identified in which affected members present with necrotizing fasciitis or severe soft tissue inflammations. Exome sequencing reveals truncating monoallelic loss-of-function variants of nuclear factor κ light-chain enhancer of activated B cells (NFKB1) in affected patients. In patients' macrophages and in NFKB1-variant-bearing THP-1 cells, activation increases both interleukin (IL)-1ß secretion and IFN-I signaling. Truncation of NF-κB1 impairs autophagy, accompanied by the accumulation of reactive oxygen species and reduced degradation of inflammasome receptor nucleotide-binding oligomerization domain, leucine-rich repeat-containing protein 3 (NLRP3), and Toll/IL-1 receptor domain-containing adaptor protein inducing IFN-ß (TRIF), thus leading to combined excessive inflammasome and IFN-I activity. Many of the patients respond to anti-inflammatory treatment, and targeting IL-1ß and/or IFN-I signaling could represent a therapeutic approach for these patients.

Nickel ion release and surface analyses on instrument fragments fractured beyond the apex: a laboratory investigation.

BACKGROUND: To analyse the changes in surface and nickel ion release characteristics of fractured root canal shaping instruments in a simulated body fluid environment. METHODS: A total of 54 new instruments were studied. The instrument groups consisted of five different NiTi alloys and a stainless-steel alloy. To standardize instrument fracture, a torsional type of failure was created on each instrument. The fractured specimens of each instrument group were randomly divided into three static immersion subgroups of 1 h, 7-day, and 30-day (n = 3). Simulated body fluid (SBF) was prepared to mimic human blood plasma by Kokubo&Takadama protocol for ex situ static immersions at 37ºC. The surfaces were examined via scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. To determine the quantitative ion release, the retrieved SBFs were analyzed using inductively coupled plasma mass spectrometry. Two-way ANOVA and Tukey post hoc tests sought the statistical significance of the nickel ion values(p < 0.05). RESULTS: In 1 h of immersion, the newly formed structures, exhibiting mostly oxygen signals, were widespread and evident on NiTi surfaces. In contrast, fewer structures were detected on the SS surface in that subgroup. In 7 days of immersion, a tendency for a decrease in the density of the new structures was revealed in NiTi groups. The oxygen signals on NiTi group surfaces significantly increased, contrary to their decrease in SS. Signals of sodium, chlorine, and calcium were detected, indicating salt precipitates in groups. In 30 days of immersion, salt precipitates continued to form. The Ni-ion release values in all instrument groups presented significant differences in comparison to the SBF control in all immersion periods(p < 0.001). No significant differences were observed in immersion time periods or instrument groups(p > 0.05). CONCLUSIONS: Within the limitations of the presented study, it was concluded that the fractured SS and NiTi root canal instruments release Ni ions in contact with body fluid. However, the Ni ion release values determined during the observation periods are lower than the critical toxic or allergic thresholds defined for the human body. This was due to the ionic dissolution cycle reaching a stable state from 1-hour to 30-day exposure to the body fluid of fractured instruments.

Evaluation of removal efficiency of capping materials used in pulp revascularization in vitro.

BACKGROUND: This study aimed to evaluate the removal efficiency of different capping materials used in pulp revascularization (PR) in a failure scenario. METHODS: The apices of freshly extracted 30 maxillary incisors were cut to mimic the immature teeth; then, root canals were shaped up to #6 Peeso reamers. The regeneration steps of the American Association of Endodontists (AAE) were followed to simulate PR treatment in vitro. The canals were dressed with the Ciprofloxacin and Metronidazole medicament mixture for 2 weeks. Then capping material groups were created: BioDentine (BD), ProRootMTA (PMTA), and RetroMTA (RMTA) (n = 10). The sealed specimens were stored for 2 weeks at 37 ºC in phosphate-buffered saline then the samples were examined by micro-computed tomography (µ-CT) analysis. Set capping materials were retrieved using a specific cement removal kit by a single blind operator. The residue materials were examined again by µCT. Kruskal-Wallis and Mann-Whitney U tests sought the significance for residue volumes. One-way ANOVA and Tukey post hoc tests with the Bonferroni corrections sought significance for the duration (p = 0.05). RESULTS: In the first examined µCT data, the mean (SD) capping material volumes of the PMTA, BD, and RMTA were 6.447 µm3 (1.086), 8.771 µm3 (0.491), and 8.114 µm3 (2.447), respectively. In the last examined µCT data, the median (IQR) residual volumes of the PMTA, BD, and RMTA were 0.051 µm3 (0.1), 0.313 µm3 (0.5), and 0.124 µm3 (0.1), respectively. A significant difference was found between BD and PMTA in the residual volumes (p < 0.05). The mean (SD) durations of the retrieving procedures of PMTA, BD, and RMTA were 19.83 min (2.34), 19.24 (3.60), and 22.04 (1.68), respectively (p = 0.063). CONCLUSIONS: Within the limitations of the presented study, it was concluded that the capping materials were largely removed from the root canals using a non-invasive approach. Nevertheless, this duration of the retrieving could be described as long.

Detection of tooth numbering, frenulum attachment, gingival overgrowth, and gingival inflammation signs on dental photographs using convolutional neural network algorithms: a retrospective study.

OBJECTIVES: This study aimed to develop an artificial intelligence (AI) model that can determine automatic tooth numbering, frenulum attachments, gingival overgrowth areas, and gingival inflammation signs on intraoral photographs and to evaluate the performance of this model. METHOD AND MATERIALS: A total of 654 intraoral photographs were used in the study (n = 654). All photographs were reviewed by three periodontists, and all teeth, frenulum attachment, gingival overgrowth areas, and gingival inflammation signs on photographs were labeled using the segmentation method in a web-based labeling software. In addition, tooth numbering was carried out according to the FDI system. An AI model was developed with the help of YOLOv5x architecture with labels of 16,795 teeth, 2,493 frenulum attachments, 1,211 gingival overgrowth areas, and 2,956 gingival inflammation signs. The confusion matrix system and ROC (receiver operator characteristic) analysis were used to statistically evaluate the success of the developed model. RESULTS: The sensitivity, precision, F1 score, and AUC (area under the curve) for tooth numbering were 0.990, 0.784, 0.875, and 0.989; for frenulum attachment these were 0.894, 0.775, 0.830, and 0.827; for gingival overgrowth area these were 0.757, 0.675, 0.714, and 0.774; and for gingival inflammation sign 0.737, 0.823, 0.777, and 0.802, respectively. CONCLUSION: The results of the present study show that AI systems can be successfully used to interpret intraoral photographs. These systems have the potential to accelerate the digital transformation in the clinical and academic functioning of dentistry with the automatic determination of anatomical structures and dental conditions from intraoral photographs.

Severity and progression rate of periodontitis are associated with an increased risk of hypertension of patients attending a university clinic.

BACKGROUND: Although periodontitis is associated with increased risk of hypertension, studies based on new periodontal disease classification is limited. We investigated whether periodontitis severity and progression rate are linked with self-reports on doctor-diagnosed hypertension in a large cohort of patients attending the periodontology clinic at the faculty of dentistry. METHODS: Archived patient files, including radiographic image records and results from full-mouth clinical periodontal examination were screened for inclusion. Data on socioeconomic factors, smoking and oral hygiene habits, and medical history were collected with a questionnaire. RESULTS: Diagnosis and background data were available for 7008 patients. The median (IQR) age was 31.0 (21.0) years; 60.1% (n = 4211) were female. Hypertension was diagnosed in 6.2% (n = 435) of patients. Both periodontitis stage and grade differed (p < 0.001) between patients with or without hypertension. Increased periodontal disease severity was associated with a 20% increasing risk for hypertension; the odds ratio (OR) was 2.63 (95% confidence interval [CI] 1.48-4.68, p < 0.001) in stage IV periodontitis. Increasing periodontitis progression rate was associated with a 35% increased risk for hypertension; the OR was 2.22 (95% CI 1.45-3.40, p < 0.001) in grade C periodontitis. CONCLUSION: Severity and progression rate of periodontitis may be independent risk factors for hypertension in this large cohort of patients attending the university periodontal department.

The impact of smoking on periodontal status and dental caries.

INTRODUCTION: Investigations to explore the relationship between smoking and its oral manifestations are important to clinicians. Among these oral manifestations, periodontal diseases and dental caries have still a controversial association. This study aims to analyze the effect of smoking on periodontal disease and caries and their relevance to each other. METHODS: Data on demographic and clinical features were retrieved from 7028 patients. Smoking status was categorized as a smoker, non-smoker, former smoker and passive smoker. Each patient received a diagnosis according to the new classification system for periodontal disease, in which periodontal disease is divides into stages (PS). The carries status was diagnosed by evaluating the decayed, missing, and filled teeth (DMFT) index. RESULTS: Of the patients, 66.6% were non-smoker women, whereas 53.7 % of passive smokers were women. Being a worker and having a Bachelor's degree was associated with a higher likelihood of getting diagnosed with periodontal disease and caries in smokers. Smoking significantly influences periodontal disease severity and DMFT values (p<0.001). This becomes more evident in former smokers by showing the highest severe periodontal problems (PS3: 29.7% and PS4: 18.9%), and the highest DMFT mean (16.4 ± 7.4) Accordingly, persons having high DMFT had significantly the most severe periodontal disease, namely PS4 (p<0.05). CONCLUSIONS: Smoking is associated with higher caries prevalence and more severe periodontal disease, and DMFT tend to increase with the severity of periodontitis in the same subjects.

The impact of smoking on oral health and patient assessment of tobacco cessation support from Turkish dentists.

INTRODUCTION: Dentists are in a critical position to help patients quit smoking. This study analyses the effectiveness of Turkish dentists in smoking cessation as part of routine patient care. METHODS: An in-person cross-sectional survey on previous dental visit experiences was completed by 226 patients recruited from the Department of Periodontology, Eskisehir Osmangazi University, Turkey, from March 2019 to September 2019. The questionnaire included topics on patient's smoking/quit characteristics, experiences on smoking cessation from their dentists, and willingness for the implementation of smoking cessation advice by dentists. RESULTS: In all, 38% of the patients were current smokers, 8% were former smokers, and 68% tried to quit previously. Smokers demonstrated consistently higher scores for plaque index, gingival index, and probing depth, than former/non-smokers (p<0.05). Patients' knowledge of adverse effects was high, and the patients presented a positive attitude toward receiving cessation activities from dentists (86.7%). A total of 89% responded positively to be asked about their smoking behavior. However, the dentists' approach for cessation discussions did not go any further than listing the harmful effects. Only 32% of the patients were informed about side effects of smoking and one-third were encouraged to quit. In general, offering smoking cessation advice was relatively infrequent, and the majority of patients tried to quit smoking by themselves (76%) without using any nicotine replacement product (84%). CONCLUSIONS: Smoking leads to oral health problems. Dentists in Turkey may ask their patients' about their smoking habits but less frequently offer practical help to quit.

Aggregatibacter actinomycetemcomitans Biofilm Reduces Gingival Epithelial Cell Keratin Expression in an Organotypic Gingival Tissue Culture Model.

Epithelial cells express keratins, which are essential for the structural integrity and mechanical strength of the cells. In the junctional epithelium (JE) of the tooth, keratins such as K16, K18, and K19, are expressed, which is typical for non-differentiated and rapidly dividing cells. The expression of K17, K4, and K13 keratins can be induced by injury, bacterial irritation, smoking, and inflammation. In addition, these keratins can be found in the sulcular epithelium and in the JE. Our aim was to estimate the changes in K4, K13, K17, and K19 expression in gingival epithelial cells exposed to Aggregatibacter actinomycetemcomitans. An organotypic gingival mucosa and biofilm co-culture was used as a model system. The effect of the biofilm after 24 h was assessed using immunohistochemistry. The structure of the epithelium was also studied with transmission electron microscopy (TEM). The expression of K17 and K19, as well as total keratin expression, decreased in the suprabasal layers of epithelium, which were in close contact with the A. actinomycetemcomitans biofilm. The effect on keratin expression was biofilm specific. The expression of K4 and K13 was low in all of the tested conditions. When stimulated with the A. actinomycetemcomitans biofilm, the epithelial contact site displayed a thick necrotic layer on the top of the epithelium. The A. actinomycetemcomitans biofilm released vesicles, which were found in close contact with the epithelium. After A. actinomycetemcomitans irritation, gingival epithelial cells may lose their resistance and become more vulnerable to bacterial infection.

The effect of smoking on the marginal bone loss around implant-supported prostheses.

INTRODUCTION: Implantology has led to several changes in the planning process involved in the application of dental prostheses to diminish bone level changes along the margins of dental implants. However, the relationship between smoking and marginal bone loss around dental implants, supporting both fixed and removable prostheses has not been investigated. We hypothesize that the design of different prostheses alter the effects of smoking, which consequently affects the amount of supporting alveolar bone. METHODS: In this study, we included 137 implants in the 'implant-supported fixed prostheses' (ISFP) group (31 smokers, 106 non-smokers) and 94 implants (21 smokers, 73 non-smokers) in the 'implant-supported removable prostheses' (ISRP) group. The corresponding patients were examined in routine recall sessions conducted at 6, 12 and 24 months after the placement of the dental prostheses. The recorded clinical periodontal parameters were the presence/ absence of a plaque index, bleeding index, and the probing depths. These periodontal parameters were assessed in conjunction with marginal bone level measurements. Comparative bone level measurements were obtained from radiographical images at ×20 magnification using the CorelDraw 11.0 software program. Statistical analysis was performed using the SPSS Statistical Software version 21.0. RESULTS: The overall clinical parameters were found to be poorer in smokers than in non-smokers (p<0.05). In all the groups, time-dependent bone loss was observed. However, among the patients with ISRPs, smokers were associated with significantly greater marginal bone loss compared to patients with ISFPs (p<0.05). CONCLUSIONS: In smokers with dental ISRPs, the marginal bone loss rates are likely to reach critical levels. Therefore, after the placement of prostheses, strict recall periods with a dental professional should be observed, and their guidance should be implemented in order to monitor the health of the bones around the implants.

A novel intrinsically disordered outer membrane lipoprotein of Aggregatibacter actinomycetemcomitans binds various cytokines and plays a role in biofilm response to interleukin-1ß and interleukin-8.

Intrinsically disordered proteins (IDPs) do not have a well-defined and stable 3-dimensional fold. Some IDPs can function as either transient or permanent binders of other proteins and may interact with an array of ligands by adopting different conformations. A novel outer membrane lipoprotein, bacterial interleukin receptor I (BilRI) of the opportunistic oral pathogen Aggregatibacter actinomycetemcomitans binds a key gatekeeper proinflammatory cytokine interleukin (IL)-1ß. Because the amino acid sequence of the novel lipoprotein resembles that of fibrinogen binder A of Haemophilus ducreyi, BilRI could have the potential to bind other proteins, such as host matrix proteins. However, from the tested host matrix proteins, BilRI interacted with neither collagen nor fibrinogen. Instead, the recombinant non-lipidated BilRI, which was intrinsically disordered, bound various pro/anti-inflammatory cytokines, such as IL-8, tumor necrosis factor (TNF)-α, interferon (IFN)-γ and IL-10. Moreover, BilRI played a role in the in vitro sensing of IL-1ß and IL-8 because low concentrations of cytokines did not decrease the amount of extracellular DNA in the matrix of bilRI- mutant biofilm as they did in the matrix of wild-type biofilm when the biofilms were exposed to recombinant cytokines for 22 hours. BilRI played a role in the internalization of IL-1ß in the gingival model system but did not affect either IL-8 or IL-6 uptake. However, bilRI deletion did not entirely prevent IL-1ß internalization, and the binding of cytokines to BilRI was relatively weak. Thus, BilRI might sequester cytokines on the surface of A. actinomycetemcomitans to facilitate the internalization process in low local cytokine concentrations.

Chronic sinusitis associated with the use of unrecognized bone substitute: a case report.

Bone grafts are used for bone augmentation to ensure optimal implant placement. However, this procedure may sometimes cause sinusitis. The case of a 44-year-old woman with the diagnosis of recurrent and chronic sinusitis of her right maxillary sinus with a history of dental implant surgery is presented. After several attempts with normal standard sinusitis therapy, unrecognized bone substitute was removed from the sinus cavity, which finally led to resolution of the sinusitis. This case reiterates the importance of a careful examination, consultation, and second opinion for the selection of optimal treatment.

A possible CD1a Langerhans cell-mast cell interaction in chronic hyperplastic candidosis.

AIMS: T lymphocyte-antigen-presenting cell (APC) interaction plays a central role in T lymphocyte activation and APC maturation. We therefore studied the CD1a-positive Langerhans cells with respect to receptor activator of nuclear factor kappa B ligand (RANKL)-positive cells in chronic hyperplastic candidosis (CHC). MATERIALS AND METHODS: Tissue sections of CHC were compared with leukoplakia and healthy oral mucosa using RANKL and CD1a monoclonal antibodies in an avidin-biotin peroxidase complex protocol. Two different antigen-retrieval protocols, pepsin preincubation and Tris-EDTA heat treatment, were used. RESULTS: CD1a-positive Langerhans cells were in healthy and leukoplakia epithelium found in the middle layer, but in CHC in all layers of the epithelium, at the basement membrane and as mononuclear round cells in the lamina propria. Use of pepsin digestion enabled studies of mast cells and their activation in the form of degranulation of RANKL. CONCLUSIONS: The numerical, morphological and topographical versatility of the CD1a-positive Langerhans cells in CHC can be clarified by dendritic cell (DC) recruitment into the epithelium. RANK-positive and RANKL-sensitive DCs have ample opportunity to interact with local T lymphocytes. Use of an optimized antigen-retrieval protocol enabled demonstration of an active engagement (degranulation) of mast cells, which represent a rapidly available source of soluble RANKL.

Effects of two multi-step self-etch primer/adhesives on apoptosis in human gingival fibroblasts in vitro.

Various in vitro studies have shown induction of apoptosis by monomers incorporated to dental restorative materials and adhesive resins, while information regarding the effect of monomer combinations as commercially available products on apoptosis is limited. The aim of this study was to investigate the effects of two multi-step self-etch primer/adhesive systems on apoptosis of cultured primary human gingival fibroblasts. Cells were treated up to 48 h with Clearfil SE Bond (Kuraray, Japan) and FL Bond (Shofu, Japan) at 1:1000 v:v ratio to determine cell proliferation, using 0.02 mM staurosporine as positive control. Apoptosis was assessed using propidium iodide/acridine orange (PI/AO) staining, compared to nontreated controls. When compared to FL Bond, exposure of gingival fibroblasts to Clearfil SE Primer and Clearfil SE Bond resulted in a higher degree of cell proliferation. PI/AO staining revealed typical morphological features of apoptosis in FL Bond and Staurosporine groups, while some cells cultured in the presence of primer and adhesive components of Clearfil SE Bond showed nuclear fragmentation, indicative of early apoptosis. Our results indicate that apoptotic potential of the multi-step self-etch adhesives were material-dependent within the 48 h test period.

Collagenases in gingival crevicular fluid in type 1 diabetes mellitus.

BACKGROUND: Studies have demonstrated that high levels of collagenase activity in gingival crevicular fluid (GCF) are associated with degradation of periodontal tissues in progressive periodontitis compared to periodontally healthy tissues. Because the activation of collagenases is an important issue in periodontitis, we have studied the activation of collagenase in gingival crevicular fluid samples of diabetic patients. METHODS: Collagenase activity was studied in human gingival crevicular fluids. Twenty-two poorly controlled diabetic patients (e.g., blood glucose: 11.0+/-0.7 mmol/l; hemoglobin A1c [HbA1c]: 9.6%+/-0.3%) and five well-controlled diabetic patients were compared to six chronic periodontitis subjects and five healthy controls. Collagenase activity against type I collagen was measured using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis quantitated by laser densitometry. RESULTS: The poorly controlled diabetic patients had more alveolar bone loss than the well-controlled diabetic subjects and controls (P<0.001; t test). The activity of collagenases in GCF in poorly controlled diabetic patients was similar to that seen in chronic periodontitis subjects (P>0.05) but higher than in healthy controls (P<0.01; t test), whereas there was no difference between the well-controlled diabetic subjects and systemically healthy controls (P>0.05; t test). CONCLUSION: Poorly controlled diabetes is strongly related to periodontal tissue destruction, and collagenases in GCF may mediate and reflect this effect.

The microenvironment around total hip replacement prostheses.

The metal stem of the totally replaced hip carries load and resists fatigue, but it is electrochemically corroded. Metallic atoms act as haptens, induce type 1 T-helper cells/Th1-type immune responses and enhance periprosthetic osteolysis. Stiff metal implants, which do not have the same elasticity as the surrounding bone, cause stress shielding. Cyclic loading and lack of ligamentous support lead to mechanical and ischemia reperfusion injury and particle formation from bone, polymethylmethacrylate, and porous implant surfaces, which accelerate third-body polyethylene wear. Surgical injury and micromotion induce the formation of a fibrous capsule interface. Type-B lining cells produce lubricin and surface-active phospholipids to promote solid-to-solid lubrication but may loosen the implant from bone. The pumping action of the cyclically loaded joint and synovial fluid pressure waves dissect the implant-host interface and transports polyethylene particles and pro-inflammatory mediators to the interface. Hyaluronan induces formation of a synovial lining like layer. Because of its localization close to bone, foreign body inflammation at the interface stimulates osteoclastogenesis and peri-implant bone loss. Metal-on-metal and ceramic-on-ceramic pairs might minimize third body wear, but can lead to high-impact load of the acetabulum. Diamond coating of a metal-on-polyethylene couple might solve both of these problems. The basic biomaterial solutions allow good mechanical performance and relatively long life in-service, but surface modifications (porous coating, hydroxyapatite, diamond, bioglass, and others) may facilitate performance of the implant and improve the biomaterial and body interfaces.