Mapping of DNA methylation-sensitive cellular processes in gingival and periodontal ligament fibroblasts in the context of periodontal tissue homeostasis.

Interactions between gingival fibroblasts (GFs) and oral pathogens contribute to the chronicity of inflammation in periodontitis. Epigenetic changes in DNA methylation are involved in periodontitis pathogenesis, and recent studies indicate that DNA methyltransferase (DNMT) inhibitors may protect against epithelial barrier disruption and bone resorption. To assess the impact of DNMT inhibition on GFs, cells were cultured with decitabine (5-aza-2'-deoxycytidine, DAC) for 12 days to induce DNA hypomethylation. We observed several potentially detrimental effects of DAC on GF biological functions. First, extended treatment with DAC reduced GF proliferation and induced necrotic cell death. Second, DAC amplified Porphyromonas gingivalis- and cytokine-induced expression and secretion of the chemokine CCL20 and several matrix metalloproteinases (MMPs), including MMP1, MMP9, and MMP13. Similar pro-inflammatory effects of DAC were observed in periodontal ligament fibroblasts. Third, DAC upregulated intercellular adhesion molecule-1 (ICAM-1), which was associated with increased P. gingivalis adherence to GFs and may contribute to bacterial dissemination. Finally, analysis of DAC-induced genes identified by RNA sequencing revealed increased expression of CCL20, CCL5, CCL8, CCL13, TNF, IL1A, IL18, IL33, and CSF3, and showed that the most affected processes were related to immune and inflammatory responses. In contrast, the genes downregulated by DAC were associated with extracellular matrix and collagen fibril organization. Our observations demonstrate that studies of DNMT inhibitors provide important insights into the role of DNA methylation in cells involved in periodontitis pathogenesis. However, the therapeutic potential of hypomethylating agents in periodontal disease may be limited due to their cytotoxic effects on fibroblast populations and stimulation of pro-inflammatory pathways.

Impact of dietary habits on the incidence of oral diseases.

BACKGROUND: Diet is a factor that can modify the course of caries, dental erosion and periodontal diseases. OBJECTIVES: The aim of this study was to examine the impact of dietary habits and the anthropometric parameters on oral health. MATERIAL AND METHODS: 50 females and 45 males aged 19-21 years were examined in a cross-sectional study. Oral health was assessed utilizing selected dental indices: approximal plaque index (API), bleeding on probing (BoP), community periodontal index (CPI), and DMFT (D - decayed, M - missing, F - filled, T - teeth). In addition, dental erosion was assessed. Anthropometric measurements included body mass, height, body mass index (BMI), waist circumference, skinfold thickness, fatty and lean body mass, body fat percentage, and total body water. The frequency of consumption of food products was determined with the use of the Questionnaire on Food Products Frequency Intake. Student's t tests, the χ2 tests and Pearson's correlation coefficients were used to analyze the results. RESULTS: The average DMFT was 9.92, API was 52.97% and BoP was 20.46%. Dental erosion was observed in 44.21% of cases. A total of 11.58% of the study population were classified as CPI 0, 30.53% as CPI 1 and 57.89% as CPI 2. The consumption of crisps and cereal products increased caries (p = 0.003). Dental erosion was associated with the consumption of fruit, vegetables, meat, fish, and alcoholic beverages. The consumption of sugar, sweets and alcoholic beverages increased API and BoP. Caries rarely occurred in people who ate fruit and vegetables on a daily basis. The anthropometric parameters were associated with oral hygiene, gingivitis and body weight disorders (p < 0.05). CONCLUSIONS: Rational nutrition not only plays a role in the development of general systemic diseases, but also has an effect on oral health. Besides providing instructions on oral hygiene, dentists should also assess the eating habits of their patients.

hTERT-immortalized gingival fibroblasts respond to cytokines but fail to mimic primary cell responses to Porphyromonas gingivalis.

In periodontitis, gingival fibroblasts (GFs) interact with and respond to oral pathogens, significantly contributing to perpetuation of chronic inflammation and tissue destruction. The aim of this study was to determine the usefulness of the recently released hTERT-immortalized GF (TIGF) cell line for studies of host-pathogen interactions. We show that TIGFs are unable to upregulate expression and production of interleukin (IL)-6, IL-8 and prostaglandin E2 upon infection with Porphyromonas gingivalis despite being susceptible to adhesion and invasion by this oral pathogen. In contrast, induction of inflammatory mediators in TNFα- or IL-1ß-stimulated TIGFs is comparable to that observed in primary GFs. The inability of TIGFs to respond directly to P. gingivalis is caused by a specific defect in Toll-like receptor-2 (TLR2) expression, which is likely driven by TLR2 promoter hypermethylation. Consistently, TIGFs fail to upregulate inflammatory genes in response to the TLR2 agonists Pam2CSK4 and Pam3CSK4. These results identify important limitations of using TIGFs to study GF interaction with oral pathogens, though these cells may be useful for studies of TLR2-independent processes. Our observations also emphasize the importance of direct comparisons between immortalized and primary cells prior to using cell lines as models in studies of any biological processes.

Oral non-specific lesions in patient with Crohn's disease - a case report.

Crohn's disease (CD) is a chronic and granulomatous inflammatory disease of the entire gastrointestinal tract. The etiopathogenesis is not fully elucidated. The most common symptoms in the active phase of the disease include abdominal pain, prolonged diarrhea, fever, fatigue, malaise and weight loss. Oral manifestations of CD are classified into specific for CD with granulomatous changes and non-specific ones. This rare extraintestinal manifestation of CD in adults may precede gastrointestinal tract involvement, occur together or appear after years of its duration. Oral lesions can be initiated by malnourishment, poor absorption of nutrients or side-effect of medications. A CASE REPORT: We describe a 28-year-old female with a 9-years CD history, who presented in the active disease with oral lesions. They were classified as non-specific ones, and included oral candidiasis, irregular erythematous patches on the cheek mucosa, exfoliative lip inflammation, and angular cheilitis. The patient was treated with azathioprine, and since the last exacerbation of symptoms, induction therapy with adalimumab, (anti-TNF-alpha), has been prescribed. Nystatin was applied to treat the oral lesions, based on the microbiological assessment of the Candida albicans susceptibility, and symptomatic treatment. After a two-week treatment the oral mucosa was healed and angular cheilitis showed marked improvement compared to the initial presentation. CONCLUSIONS: The young female with active CD presented the nonspecific lesions in the oral cavity. The lesions coexisted with the active inflammatory process in the intestinal tract with characteristic clinical symptoms, and were associated with sideropenic anemia. The implementation of the local therapy, systemic CD treatment and supplementation of micronutrient deficiencies have led to a healing of the oral lesions. We emphasize a personalized approach to treatment and close cooperation between the dentist and the gastroenterologist.

Evaluation of Proinflammatory, NF-kappaB Dependent Cytokines: IL-1α, IL-6, IL-8, and TNF-α in Tissue Specimens and Saliva of Patients with Oral Squamous Cell Carcinoma and Oral Potentially Malignant Disorders.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a life-threatening disease. It could be preceded by oral potentially malignant disorders (OPMDs). It was confirmed that chronic inflammation can promote carcinogenesis. Cytokines play a crucial role in this process. The aim of the study was to evaluate interleukin-1alpha (IL-1α), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-α) in tissue specimens and saliva of patients with OSCC and OPMDs. METHODS: Cytokines were evaluated in 60 tissue specimens of pathological lesions (OSCCs or OPMDs) and in 7 controls (normal oral mucosa, NOM) by immunohistochemistry and in saliva of 45 patients with OSCC or OPMDs and 9 controls (healthy volunteers) by enzyme-linked immunosorbent assays. RESULTS: Immunohistochemical analysis revealed significantly higher expression of IL-8 in OSCC specimens and TNF-α in OSCCs and OPMDs with dysplasia as compared to NOM. Moreover, expression of TNF-α was significantly higher in oral leukoplakia and oral lichen planus without dysplasia, whereas expression of IL-8 only in oral leukoplakia without dysplasia in comparison with NOM. Salivary concentrations of all evaluated cytokines were significantly higher in patients with OSCC than in controls. Moreover, levels of IL-8 were significantly higher in saliva of patients with OPMDs with dysplasia as compared to controls and in OSCC patients as compared to patients with dysplastic lesions. There was also significant increase in salivary concentrations of IL-6, IL-8 and TNF-α in patients with OSCC as compared to patients with OPMDs without dysplasia. CONCLUSION: The study confirmed that proinflammatory, NF-kappaB dependent cytokines are involved in pathogenesis of OPMDs and OSCC. The most important biomarker of malignant transformation process within oral mucosa among all assessed cytokines seems to be IL-8. Further studies on a larger sample size are needed to corroborate these results.

The Relationship between the Concentration of Salivary Tyrosine and Antioxidants in Patients with Oral Lichen Planus.

The diagnosis of oral lichen planus (OLP) is based on clinical examination and histopathological criteria. Noninvasive diagnostics of saliva may be considered as a confirmation of OLP diagnosis and a potential alternative to an invasive method. The objective of the present study was to evaluate the relationship between the level of tyrosine (Tyr) as well as antioxidants like uric acid (UA) and glutathione peroxidase (GPx) activity in the saliva of patients with OLP in comparison with the control group (healthy subjects without any oral changes). A total of 40 patients with OLP and 40 healthy volunteers were selected for the study based on the modified WHO diagnostic (clinical and histopathological) criteria. High-performance liquid chromatography (HPLC) was performed for Tyr concentration, while GPx activity and uric acid levels were determined by a colorimetric method. The concentrations of Tyr, UA, and GPx activity were statistically lowered in OLP patients compared to the control group. All examined parameters correlated strongly and positively with each other. Mean values of salivary UA concentrations differed between the groups of OLP patients (reticular and erosive forms) and controls (206.66 vs. 196.54 vs. 218.49 µmol/L, respectively, p = 0.001). A similar trend was demonstrated in salivary Tyr concentration which differed statistically between the study and control groups (0.08 vs. 0.07 vs. 0.13 µmol/L, respectively, p = 0.001). Determining of a relationship between the concentrations of Tyr, UA, and GPx activity may be useful in the prognosis of OLP. The HPLC method may be employed, as an additional noninvasive diagnostic procedure to screen OLP patients, during the routine diagnostics of salivary biochemical parameters such as aromatic amino acids.

Validation of a modified Oral Health Impact Profile scale (OHIP-14) in patients with oral mucosa lesions or periodontal disease.

BACKGROUND: Now that healthcare systems have helped successfully extend the human lifespan, the next challenge is to improve the patient's quality of life (QOL), in particular health-related quality of life (HRQOL). A proper HRQOL assessment requires using reliable instruments that are well-adapted to the population. OBJECTIVES: The objective of this study was to validate a modified Polish version of the Oral Health Impact Profile scale (OHIP-14) for patients with oral mucosa lesions or periodontal disease. MATERIAL AND METHODS: The sample consisted of 180 adults seeking highly specialized treatment at the Periodontology Department of the University Dental Clinic in Kraków, Poland. The main modification made to OHIP-14 was the inclusion of subquestions regarding the teeth (subscale 1), oral mucosa and other soft tissues (subscale 2), and dentures (subscale 3). RESULTS: The Cronbach's alpha values were excellent for all 3 subscales (subscale 1: α = 0.924; subscale 2: α = 0.937; subscale 3: α = 0.936). In the case of subscale 1, the Kaiser criterion showed a model with 3 factors ("psychological and social limitations"; "physical limitations"; "functional limitations"), which together explained 67.1% of the variance, in the case of subscale 2 - a model with 1 factor, and in the case of subscale 3 - a 2-factor model ("social interactions limitations"; "basic activities disorder and personal discomfort"). CONCLUSIONS: Statistical testing demonstrated that a modified OHIP-14 questionnaire is a reliable tool for evaluating QOL in patients with periodontal or oral mucosa diseases.

Oral manifestations of granulomatosis with polyangiitis - Clinical and radiological assessment.

BACKGROUND/PURPOSE: Granulomatosis with polyangiitis (GPA) is a type of primary systemic vasculitis associated with the presence of anti-neutrophil cytoplasmic antibodies (ANCA). GPA mainly affects medium and small vessels and may manifest in different organs, most commonly upper respiratory tract. Oral lesions occur in 6-13% of GPA patients and might be the first symptom observed by the patient. This study presents the evaluation of orofacial manifestations of patients with GPA. MATERIALS AND METHODS: Prospective study was conducted between June 2014 and November 2017 in Department of Allergy and Immunology at University Hospital in Krakow. Patients diagnosed with GPA, after medical assessment, were examined including precise dental inspection, bacterial and fungal flora of oral cavity evaluation and Cone Beam Computed Tomography (CBCT) imaging. RESULTS: Nine patients were enrolled in the study. Characteristic for GPA strawberry gingivitis was observed in one patient. Bone destruction and inflammatory lesions in paranasal sinuses was confirmed by CBCT in 55.6% of patients. Fungal infection was revealed in 66.7% of patients. CONCLUSION: These findings oblige dentists to consult patient with laryngologist or internal medicine physician to establish further diagnostic approach, because early diagnosis of GPA is crucial for implementing appropriate treatment and preventing chronic organ damage.

Evaluation of enzymatic and non-enzymatic antioxidant status and biomarkers of oxidative stress in saliva of patients with oral squamous cell carcinoma and oral leukoplakia: a pilot study.

Objective: The study aimed to evaluate total antioxidant capacity as well as levels of various enzymatic and non-enzymatic antioxidants, and oxidative stress markers in saliva of patients with oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL). Material and methods: Twenty patients with OSCC, 20 patients with OL and 20 healthy subjects were enrolled into this prospective study. Total Antioxidant Capacity (TAC), activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) as well as levels of total glutathione (tGSH), reduced glutathione (GSH), oxidized glutathione (GSSG), GSH/GSSG ratio, uric acid (UA), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) were estimated in saliva using appropriate biochemical methods. Results: The activity of SOD was significantly higher in OSCC group in comparison with OL and control groups. The levels of GSH were markedly lower in OSCC and OL patients as compared to the control group. Likewise, we found that GSH/GSSG ratio was markedly lower in the OSCC and OL groups. Levels of some biomarkers were influenced by clinical staging of OSCC and OL as well as by sociodemographic factors. Conclusions: The results of this pilot study suggest that salivary activity of SOD is higher in OSCC patients, whereas levels of GSH and GSH/GSSG ratio are lower in saliva of patients with OSCC and OL. Clinical staging of OSCC and OL, as well as some sociodemographic factors may also influence salivary antioxidant status.

[Unilateral idiopathic gingival fibromatosis--a case report]. / Jednostronny idiopatyczny przerost dziasel--opis przypadku.

Gingival fibromatosis is a painless gingival overgrowth. It may result in difficulties with proper dental hygiene keeping, mastication and occlusion. Herein, a case of a 10-year-old patient was described. The patient reported to the Department of Periodontology and Oral Medicine of the Jagiellonian University Medical College in Krakow due to the problems with permanent teeth eruption (23-26), chewing and dental hygiene maintaining. Based on medical history, clinical examination, diagnostic tests and histopathological study of gingival tissue biopsies the patient was diagnosed with unilateral idiopathic gingival fibromatosis. After oral cavity hygienization, patient un- derwent dental surgery procedures by gingivectomy and gingivoplasty. The follow-up examination 2 and 6 months post operation showed un- eventful healing, proper tooth eruption, improved oral hygiene and chewing function. Twelve months post surgery no recurrence was noted. Due to the etiological diversity of gingival lesions occurring as an overgrowth, accurate medical history, clinical examination, laboratory tests and histopathological study are needed. Accurate diagnos- tics is crucial mainly to exclude he- matological and oncological diseases. Gingivectomy being the "gold method" of gingival fibromatosis treatment was effective and sufficient to cure the case presented in this article.

Clinical crown lengthening - a case report.

Maintaining healthy parodontium during teeth restoration procedures is an indispensable condition for obtain- ing regular functionning and esthetics. Thus, the knowledge of correct anatomy and the influence of filling and complement on paradontium tissue is vital. Difficulty in maintaining appropriate gingival biological width (GBW) is a frequent problem encountered in this type of reconstruction. Preservation of unchanged biological width conditions predictible treatment result and, what is more, lack of inflammatory lesions in marginal parodontium. The ideal situation for parodontium is localizing the filling/complement border supragingivaly, which is at least 3 mm from alveolar process edge. In the case, when the above conditions are impossible to fulfil, elongation of clinical crown is a metod of choice. The effect is possible to achieve by surgical treatment or combined orthodontic - surgical treatment. Surgical treatment is faster and preferred procedure for indirect reconstruction, where achieving high clinical crown is necessary. Three clinical cases of performing method of surgical clinical crown elongation were presented in the article. Performing the described procedure enables correct tooth crown reconstruction and, what is most important, keeping individual toothing.

Interleukin-6 and neopterin levels in the serum and saliva of patients with Lichen planus and oral Lichen planus.

BACKGROUND: Lichen planus together with its oral variant is a chronic, inflammatory disease of the skin and the mucosa of unclear aetiology and with an unpredictable course that still poses a major problem in terms of diagnosis and treatment. The objective of this study was to assess the concentrations of interleukin-6 (IL-6) and neopterin in saliva and serum of patients with lichen planus (including reticular and erosive form of oral lichen planus) and to compare them with the concentrations observed in healthy controls. METHODS: The study material comprised serum and saliva samples from 56 patients diagnosed with lichen planus and 56 healthy volunteers. The ELISA test was used to measure concentrations of IL-6 and neopterin in the serum and saliva of the study participants. RESULTS: The concentrations of IL-6 in saliva and serum of patients with lichen planus were significantly higher than in controls (P = 0.0002; P < 0.0001). The difference remains significant after adjustment for gingivitis and age. Patients with atrophic-erosive oral lichen planus had significantly higher IL-6 concentrations in their saliva compared to patients with reticular form of disease (P = 0.01). The concentrations of neopterin were significantly higher in the serum but not in saliva of lichen planus patients vs. controls (P <0.0001). CONCLUSIONS: Serum levels of proinflammatory cytokines IL-6 and neopterin are increased in lichen planus as well as the salivary concentrations of IL-6. The differences observed in IL-6 levels in patients with erosive-atrophic forms of oral lichen planus may indicate a substantial role played by the cytokine in the disease.

[Inflammatory changes of oral cavity proceeding appearance of clinical symptoms of Crohn's disease]. / Zmiany zapalne w jamie ustnej wyprzedzajace wystapienie objawów choroby Lesniowskiego-Crohna. Opis przypadku.

Crohn's disease belongs to the inflammatory bowel diseases. Inflammatory changes can be located in any part of the gastrointestinal tract including rarely oral cavity. We present a case of a 21 years old woman with unhealed by the local treatment, and verified by histological examination, inflammatory changes of oral cavity. These changes had proceed the diagnosis of typical changes in gastrointestinal tract for 6 months. Changes were located in terminal ileum and colon and were confirmed by colonoscopic, histologic and radiologic studies. Standard therapy of Crohn's disease with antiinflammatory and immunosuppressive drugs led to the healing of oral changes. We point out on the necessity of proper differential diagnosis of problematic unhealing changes, particularly with ulcerations of oral mucosa in young patients. Dental examination with histological confirmation of mucosal changes may be helpful in proper diagnosis of Crohn's disease in the young group of patients.

[Influence of smoking on incidence and progression of periodontal disease. Review of the literature]. / Wplyw palenia tytoniu na wystepowanie i przebieg chorób przyzebia. Przeglad pismiennictwa.

On the basis of available literature, presented is the influence of smoking on incidence and progression of peridontal disease. During the process of smoking; more than 400 destructive substances along with nicotine are excreted. Their effects on peridontal tissue present as direct and indirect symptoms (due to disfunction of the immunological reactions) which permit for better conditions for pathological bacterial invasion and increased secretion of proinflammatory factors.

[Comparison of the systems used for providing local anesthesia in dentistry--the Wand (Milestone Scientific) and Injex (Rosch)]. / Analiza porównawcza systemów do znieczulen miejscowych w stomatologii--the Wand (Milestone Scientific) oraz Injex (Rosch).

UNLABELLED: Local anesthesia is one of the basic and the most often executed interventions in dentistry. This procedure is very stressful for the patients because it is combined with pain. The new systems for delivering local anesthesia in dentistry have revolutionized the technique considerably by its simplify as well as reduction in pain. THE AIM: this study presents the comparison between the local anesthesia delivery systems used in dentistry--The Wand and Injex, taking into consideration pain intensity during performing anesthesia and the intensification of fear before executed anesthesia with the given system. MATERIAL AND METHODS: the Visual Analogue Scale (VAS), verbal scale and questionnaires were used to evaluate pain and fear. RESULTS: On the basis of our investigations it can be concluded that there were statistically important differences between men and women in fear intensity combined with the anesthesia procedure--men were less afraid than women. The patients who were anaesthetized with system The WAND declared less fear before similar anesthesia in future. The average value of intensity of pain analyzed with both verbal and visual scales during anaesthetizing with the system Injex (independently from sex) was statistically significantly higher than for system The WAND--respectively 0.57 and 8.55 for The WAND, 2.02 and 32.18 for Injex (p = 0.001). CONCLUSION: on the basis of the results of this study it can be concluded that the less stressful and painful local anesthesia delivery system is the WAND.

[Mucositis--a major non-hematologic complication of high-dose chemotherapy and radiotherapy--pathogenesis, prevention and treatment]. / Mucositis--najczestsze niehematologiczne powiklanie wysokodozowanej chemio i radioterapii--patogeneza, zapobieganie, leczenie.

High-dose chemotherapy and radiotherapy--induced oral mucositis (OM) represents a major non-hematologic complication of stem cell transplantation in cancer patients. This side effect causes significant treatment-related morbidity and is an important clinical problem because of the pain, odynodysphagia, the requirement for parenteral nutrition, narcotics and the risk of local infections which are exacerbated by the severe neutropenia and can become systemic. The pathogenesis of OM can be attributed to the direct mucosal toxicity of high-dose chemotherapy and ionizing radiation and to indirect mucosal damage caused by a concomitant local bacterial, viral and mycotic infections. This review describes pathophysiology as well as prophylactic and therapeutic armamentarium for the treatment of oral mucositis in patients receiving high dose chemotherapy and/or radiotherapy.