Identification of a new genetic variant (G231N, E232T, N235D) of peptidylarginine deiminase from P. gingivalis in advanced periodontitis.

Chronic periodontitis (CP), an inflammatory disease of periodontal tissues driven by a dysbiotic subgingival bacterial biofilm, is also associated with several systemic diseases, including rheumatoid arthritis (RA). Porphyromonas gingivalis, one of the bacterial species implicated in CP as a keystone pathogen produces peptidyl arginine deiminase (PPAD) that citrullinates C-terminal arginine residues in proteins and peptides. Autoimmunity to citrullinated epitopes is crucial in RA, hence PPAD activity is considered a possible mechanistic link between CP and RA. Here we determined the PPAD enzymatic activity produced by clinical isolates of P. gingivalis, sequenced the ppad gene, and correlated the results with clinical determinants of CP in patients from whom the bacteria were isolated. The analysis revealed variations in PPAD activity and genetic diversity of the ppad gene in clinical P. gingivalis isolates. Interestingly, the severity of CP was correlated with a higher level of PPAD activity that was associated with the presence of a triple mutation (G231N, E232T, N235D) in PPAD in comparison to W83 and ATCC 33277 type strains. The relation between mutations and enhanced activity was verified by directed mutagenesis which showed that all three amino acid residue substitutions must be introduced into PPAD expressed by the type strains to obtain the super-active enzyme. Cumulatively, these results may lead to the development of novel prognostic tools to assess the progress of CP in the context of associated RA by analyzing the ppad genotype in CP patients infected with P. gingivalis.

Salivary matrix metalloproteinase-9 (MMP-9) as a biomarker of periodontitis in pregnant patients with diabetes.

BACKGROUND: Pregnancy and diabetes increase the risk of developing pathological conditions in the periodontium. Salivary biomarkers, such as matrix metalloproteinase-9 (MMP-9), as well as antioxidants can be used as diagnostic indicators in monitoring periodontitis. OBJECTIVES: The aim of the study was to assess the periodontal status of pregnant women with regard to the presence of diabetes. In addition, we aimed to assess antioxidant activity and the level of MMP-9 in saliva in order to establish the optimal noninvasive determinants of periodontitis. MATERIAL AND METHODS: The study included 104 pregnant women: 35 patients had gestational diabetes mellitus (GDM); 30 patients had type 1 diabetes (T1D); and 39 patients did not have diabetes (the control group). The physical examination included the assessment of the approximal plaque index (API), the gingival index (GI), bleeding on probing (BOP), the probing pocket depth (PPD), and clinical attachment loss (CAL). In the saliva study, MMP-9 concentration as well as the ferric reducing ability of plasma (FRAP), and the activity of superoxide dismutase (SOD), glutathione reductase (GR) and glutathione peroxidase (GPX) were measured. RESULTS: The pregnant patients with GDM and T1D had higher GI, BOP, PPD, and CAL scores than the control women (p < 0.0001, p = 0.0040, p = 0.0100, p = 0.0030, and p < 0.0001, p < 0.0009, p < 0.0001, p < 0.0001, respectively). The T1D patients had higher API scores as compared to the control women (p = 0.0010). The patients with periodontitis had higher salivary MMP-9 levels than the patients without periodontitis (p = 0.0001). The salivary antioxidant levels and activity were comparable among the study groups. The determinants of periodontitis (p < 0.0001) were MMP-9 concentration (p = 0.0008) and oral hygiene (p = 0.0001). The concentration of MMP-9 was also a useful determiner of the presence of periodontitis (p < 0.0001). CONCLUSIONS: In the pregnant women with diabetes, we observed worse gingival conditions, deeper periodontal pockets and greater attachment loss in comparison with the women from the control group. The concentration of MMP-9 in saliva is a good predictor of periodontitis and might be a useful tool for diagnosing periodontitis.

Oral health-related quality of life (OHRQoL) in Polish adults with periodontal diseases, oral mucosal diseases and dental caries.

BACKGROUND: With the development of medicine and extending the human lifespan, the next challenge for healthcare providers is to improve the quality of life. Oral Health Impact Profile (OHIP) is a worldwide known questionnaire that is used for assessing oral health-related quality of life (OHRQoL). OBJECTIVES: The aim of the present study was to assess the impact of periodontal diseases, oral mucosal lesions and dental caries on OHRQoL among Polish adults. MATERIAL AND METHODS: A cross-sectional study consisting of an intraoral clinical examination and a questionnaire was conducted among 250 adult patients seeking dental treatment at the University Dental Clinic (UDC) in Cracow, Poland. The obtained clinical data included the number of decayed, filled and missing teeth (DMFT), the presence of fixed or removable dental prostheses, the type and size of oral mucosal diseases, periodontal data based on a visual examination as well as the approximal plaque index (API) and modified sulcus bleeding index (mSBI) scores, and the patient's dental history. A modified OHIP questionnaire was used, which had been previously validated amongst patients with periodontal and oral mucosal diseases. RESULTS: In patients reporting problems with oral mucosa, the OHIP-14 scores in relation to oral mucosa and other soft tissues were higher, and the scores in relation to the teeth were lower than in patients who did not suffer from oral mucosal diseases (0.86 (0.25-1.81) vs. 0.29 (0-1.00); p < 0.001, and 0.39 (0.07-1.07) vs. 0.68 (0.29-1.29); p = 0.048, respectively). Among patients looking for treatment due to caries and other dental problems, the OHIP-14 scores relating to dentures were higher and the scores relating to oral mucosa were lower than in patients who did not report such problems (2.07 (0.96-2.15) vs. 0.64 (0-1.38); p = 0.043, and 0.14 (0-0.56) vs. 0.57 (0.14-1.31); p = 0.001, respectively). Among patients noticing prosthetic problems, the OHIP-14 scores relating to dentures were higher than in those who did not suffer from such issues (2.07 (1.23-2.36) vs. 0.64 (0-1.36); p = 0.004). CONCLUSIONS: The symptoms reported by patients with periodontal diseases, oral mucosal lesions and dental caries influenced their OHRQoL. The proper prophylaxis and treatment of these diseases are important to avoid the worsening of OHRQoL.

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.

Oral Health Impact Profile in adult Polish population.

BACKGROUND: Oral Health Related Quality of Life (OHRQoL) is a significant patient-oriented outcome which should be considered in diagnostic, decision making and therapeutic process by dentists. OBJECTIVES: To evaluate the influence of a condition of teeth, oral mucosa and dentures on the OHRQoL among patients seeking dental treatment in the University Dental Clinic (UDC) in Kraków, Poland. MATERIAL AND METHODS: 250 patients took part in a cross-sectional study conducted in the UDC in Kraków. Collected clinical data included: oral mucosa and periodontal condition, number of Decayed, Missed and Filled Teeth (DMFT index), presence of dentures. Questionnaire part of the study has been based on OHIP-14 questionnaire. RESULTS: The condition of teeth worsened OHRQoL in following groups of patients: first-time visitors, smokers, with periodontal diseases and without oral mucosal diseases. There was a positive significant correlation between number of decayed teeth and OHRQoL related to teeth. The condition of oral mucosa significantly impaired OHRQoL in: women, patients with oral mucosal diseases and those who undergone prosthetic treatment in the last year. There was a correlation between number of missing teeth, DMFT index, age and subscale 2 of the form. The condition of dentures significantly impaired the OHRQoL in patients: who used removable dental prostheses, undergone prosthetic treatment in the last year and females. There was a positive correlation between subscale 3 of the questionnaire and number of missing teeth and DMFT index and a negative correlation with number of filled teeth within this subscale. CONCLUSIONS: Evaluation of patient's OHRQoL is one of important components for successful dental treatment.

Antimicrobial photodynamic therapy effectively reduces Porphyromonas gingivalis infection in gingival fibroblasts and keratinocytes: An in vitro study.

BACKGROUND: Porphyromonas gingivalis possess the ability to invade host cells which prevents this pathogen from eradication by conventional periodontal therapy. Recently, antimicrobial photodynamic therapy (aPDT) was introduced to periodontal treatment as a complementary antibacterial method. The aim of this study was to evaluate the effect of toluidine blue-O (TBO) mediated aPDT on the viability of P. gingivalis invading gingival fibroblasts and keratinocytes in an in vitro model of infection. METHODS: Primary human gingival fibroblasts (PHGF) and telomerase immortalized gingival keratinocytes (TIGK) were infected with Pg ATCC 33277. Two concentrations of TBO (0.01 mg/mL, TBO-c1 and 0.001 mg/mL, TBO-c2) and a non-laser red light source (λ = 630 nm) were applied to treat both cell-adherent/intracellular Pg (the adhesion/invasion model) or exclusively the intracellular bacteria (the intracellular infection model). RESULTS: The median viability of cell-adherent/intracellular Pg in infected keratinocytes declined from 1.88 × 105 cfu/mL in infected cells treated with TBO without irradiation to 40 cfu/mL upon irradiation for 10 s with TBO-c1. At higher light doses a complete photokilling of P. gingivalis was observed. Pg from exclusively intracellular infection model was also efficiently eradicated as the residual viability dropped from 1.44 × 105 cfu/mL in control samples to 160, 20 and 10 cfu/mL upon irradiation for 10, 20 and 30 s, respectively. In the infected fibroblasts irradiation significantly reduced bacterial viability but did not completely eradicate the intracellular pathogen. CONCLUSIONS: Antimicrobial PDT is effective in reducing the viability of intracellular periopathogens, however those residing within gingival fibroblasts seems to attenuate the photokilling effectiveness of this method.

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.

Candida albicans Shields the Periodontal Killer Porphyromonas gingivalis from Recognition by the Host Immune System and Supports the Bacterial Infection of Gingival Tissue.

Candida albicans is a pathogenic fungus capable of switching its morphology between yeast-like cells and filamentous hyphae and can associate with bacteria to form mixed biofilms resistant to antibiotics. In these structures, the fungal milieu can play a protective function for bacteria as has recently been reported for C. albicans and a periodontal pathogen-Porphyromonas gingivalis. Our current study aimed to determine how this type of mutual microbe protection within the mixed biofilm affects the contacting host cells. To analyze C. albicans and P. gingivalis persistence and host infection, several models for host-biofilm interactions were developed, including microbial exposure to a representative monocyte cell line (THP1) and gingival fibroblasts isolated from periodontitis patients. For in vivo experiments, a mouse subcutaneous chamber model was utilized. The persistence of P. gingivalis cells was observed within mixed biofilm with C. albicans. This microbial co-existence influenced host immunity by attenuating macrophage and fibroblast responses. Cytokine and chemokine production decreased compared to pure bacterial infection. The fibroblasts isolated from patients with severe periodontitis were less susceptible to fungal colonization, indicating a modulation of the host environment by the dominating bacterial infection. The results obtained for the mouse model in which a sequential infection was initiated by the fungus showed that this host colonization induced a milder inflammation, leading to a significant reduction in mouse mortality. Moreover, high bacterial counts in animal organisms were noted on a longer time scale in the presence of C. albicans, suggesting the chronic nature of the dual-species infection.

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.

Metabolomic Status of The Oral Cavity in Chronic Periodontitis.

Chronic periodontitis is an inflammatory disease of tooth-supporting tissues associated with Porphyromonas gingivalis. Expansion and invasion of this bacterium into the periodontium is associated with changes in the metabolome of the oral cavity. MATERIALS AND METHODS: Metabolomics analysis of mouth washout and tongue swab samples based on proton nuclear magnetic resonance (1H-NMR) method was employed to determine metabolic status of the oral cavity in chronic periodontal disease. RESULTS: Mouth washout extracts contained a total of 23 metabolites and tongue swab extracts contained 17. Identified metabolites partially overlap with the content of saliva and gingival crevicular fluid. The colonization of the oral cavity of patients with periodontitis by bacteria was manifested in the change in levels of eight metabolites. CONCLUSION: NMR-based metabolomics analysis is a potentially useful methodological approach for monitoring the pathological processes observed in the oral cavity in the course of periodontitis.

Evaluation of depression, anxiety and stress levels in patients with oral lichen planus.

The pathogenesis of oral lichen planus (OLP) remains to be fully elucidated; however, certain psychoneurological factors may influence the onset and exacerbation of OLP. The aim of the present study is to evaluate the intensity of negative emotions in patients with OLP. A cross-sectional, questionnaire-based study was performed. The sample consisted of 52 subjects, comprising 26 patients with OLP (OLP group) and 26 controls (CTRL group). The Depression Anxiety Stress Scale 21 (DASS-21) was used for psychometric evaluation. The patients were also asked about their attitude toward the disease, treatment, and interference of the disease on daily life. The mean level of depression was 12.54 ± 6.6 in the OLP group and 7.69 ± 5.22 in the CTRL group (P = 0.006). The mean level of anxiety was 11.15 ± 7.95 in the OLP group and 6.62 ± 4.86 in the CTRL group (P = 0.018). The mean stress levels were 8.69 ± 7.06 and 3.85 ± 3.18 in the OLP and CTRL groups, respectively (P = 0.003). Severe and very severe scores of depression and very severe scores of anxiety and stress were present in the OLP group, whereas these emotions were normal in the majority of controls. Depression, stress, and anxiety may be involved in the pathogenesis and course of OLP.

BET Bromodomain Inhibitors Suppress Inflammatory Activation of Gingival Fibroblasts and Epithelial Cells From Periodontitis Patients.

BET bromodomain proteins are important epigenetic regulators of gene expression that bind acetylated histone tails and regulate the formation of acetylation-dependent chromatin complexes. BET inhibitors suppress inflammatory responses in multiple cell types and animal models, and protect against bone loss in experimental periodontitis in mice. Here, we analyzed the role of BET proteins in inflammatory activation of gingival fibroblasts (GFs) and gingival epithelial cells (GECs). We show that the BET inhibitors I-BET151 and JQ1 significantly reduced expression and/or production of distinct, but overlapping, profiles of cytokine-inducible mediators of inflammation and bone resorption in GFs from healthy donors (IL6, IL8, IL1B, CCL2, CCL5, COX2, and MMP3) and the GEC line TIGK (IL6, IL8, IL1B, CXCL10, MMP9) without affecting cell viability. Activation of mitogen-activated protein kinase and nuclear factor-κB pathways was unaffected by I-BET151, as was the histone acetylation status, and new protein synthesis was not required for the anti-inflammatory effects of BET inhibition. I-BET151 and JQ1 also suppressed expression of inflammatory cytokines, chemokines, and osteoclastogenic mediators in GFs and TIGKs infected with the key periodontal pathogen Porphyromonas gingivalis. Notably, P. gingivalis internalization and intracellular survival in GFs and TIGKs remained unaffected by BET inhibitors. Finally, inhibition of BET proteins significantly reduced P. gingivalis-induced inflammatory mediator expression in GECs and GFs from patients with periodontitis. Our results demonstrate that BET inhibitors may block the excessive inflammatory mediator production by resident cells of the gingival tissue and identify the BET family of epigenetic reader proteins as a potential therapeutic target in the treatment of periodontal disease.

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.

Fibroblasts from recurrent fibrotic overgrowths reveal high rate of proliferation in vitro - findings from the study of hereditary and idiopathic gingival fibromatosis.

PURPOSE: Investigate the content of fibrotic fibrils in gingival tissue and the proliferation of fibroblasts collected from recurrent and non-recurrent hereditary gingival fibromatosis (HGF) and idiopathic gingival fibromatosis (IGF). METHODS: Gingival biopsies were collected from HGF (n = 3) and IGF (n = 3) donors with recurrent and non-recurrent gingival overgrowths and from a control group (Ctrl, n = 3). Hematoxylin staining was performed to evaluate the histomorphology of gingival tissue. Heidenhain's AZAN trichrome staining served for visualization of fibrotic fibrils in gingiva. Quantitative analysis of the content of fibrotic fibrils in gingival tissue was performed using a polarized light microscope. Proliferation was evaluated at 24 h, 48 h, and 72 h in fibroblast cultures using a cell proliferation ELISA assay based on 5-bromo-2'-deoxyuridine (BrdU). RESULTS: Numerous blood vessels and fibroblasts were observed in recurrent overgrowths, whereas moderate blood vessels and moderate to scanty fibroblasts were detected in non-recurrent overgrowths. Heidenhain's staining revealed numerous collagen fibers in both recurrent and non-recurrent overgrowths. Quantitative analysis in a polarizing microscope showed significant accumulation of fibrotic fibrils exclusively in the overgrowths with the recurrence. In all time-points, increased proliferation of cells from all recurrent overgrowths was observed, but not from overgrowths which do not reoccur. CONCLUSIONS: The study revealed that recurrent gingival overgrowths consist of highly fibrotic and dense connective tissue with numerous blood vessels and abundant fibroblasts. We also demonstrated that unlike fibroblasts derived from overgrowths, which did not present recurrence, fibroblasts derived from highly fibrotic and recurrent overgrowths maintain high rate of proliferation in vitro.

Intraoral manifestation of systemic AL amyloidosis with unique microscopic presentation of intracellular amyloid deposition in striated muscles.

We report the history of a 59-year old patient with systemic AL amyloidosis of intraoral manifestation. The patient first presented with complaints about dysphagia and remarkable enlargement of the tongue with highly reduced mobility, as well as bilateral submucosal thickenings on the cheeks. Histopathological examination of the incisional biopsy of the buccal mucosa and underlying tissues revealed AL amyloidosis. The microscopic presentation was, however, unique, as the amyloid deposits were present intracellularly in the striated muscles. The subsequent bone marrow biopsy confirmed the diagnosis of primary amyloidosis/multiple myeloma - associated amyloidosis.

TIMP-1 association with collagen type I overproduction in hereditary gingival fibromatosis.

OBJECTIVES: To investigate the processes associated with the excessive production of collagen I in hereditary gingival fibromatosis (HGF). MATERIALS AND METHODS: Three HGF subjects and five controls were enrolled in the study. Histomorphological and immunohistological analyses were performed on gingival tissues. The expression of heat-shock protein 47 (HSP47), collagen I, transforming growth factor-ß1 (TGF-ß1), connective tissue growth factor (CTGF), matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) by gingival fibroblasts isolated from HGF and controls was analysed using qRT-PCR, Western blotting and ELISA. RESULTS: Considerable accumulation of fibrotic fibrils and increased synthesis of HSP47 were noted in HGF gingival tissues. The synthesis of collagen I, HSP47, TGF-ß1, CTGF and TIMP-1 was significantly elevated in HGF gingival fibroblasts compared with controls, while the production of MMP-1 was decreased. CONCLUSIONS: We report that fibrosis in HGF gingival tissues is associated with increased synthesis of HSP47. This finding was confirmed by an in vitro study, where excessive production of collagen I was associated with increased synthesis of HSP47, TGF-ß1 and CTGF by HGF gingival fibroblasts. Moreover, the shift in the TIMP-1/MMP-1 ratio identifies increased synthesis of TIMP-1 as one of the processes associated with collagen I overproduction in HGF fibroblasts.

Association of Distinct Fine Specificities of Anti-Citrullinated Peptide Antibodies With Elevated Immune Responses to Prevotella intermedia in a Subgroup of Patients With Rheumatoid Arthritis and Periodontitis.

OBJECTIVE: In addition to the long-established link with smoking, periodontitis (PD) is a risk factor for rheumatoid arthritis (RA). This study was undertaken to elucidate the mechanism by which PD could induce antibodies to citrullinated peptides (ACPAs), by examining the antibody response to a novel citrullinated peptide of cytokeratin 13 (CK-13) identified in gingival crevicular fluid (GCF), and comparing the response to 4 other citrullinated peptides in patients with RA who were well-characterized for PD and smoking. METHODS: The citrullinomes of GCF and periodontal tissue from patients with PD were mapped by mass spectrometry. ACPAs of CK13 (cCK13), tenascin-C (cTNC5), vimentin (cVIM), α-enolase (CEP-1), and fibrinogen ß (cFIBß) were examined by enzyme-linked immunosorbent assay in patients with RA (n = 287) and patients with osteoarthritis (n = 330), and cross-reactivity was assessed by inhibition assays. RESULTS: A novel citrullinated peptide cCK13-1 (444 TSNASGR-Cit-TSDV-Cit-RP458 ) identified in GCF exhibited elevated antibody responses in RA patients (24%). Anti-cCK13-1 antibody levels correlated with anti-cTNC5 antibody levels, and absorption experiments confirmed this was not due to cross-reactivity. Only anti-cCK13-1 and anti-cTNC5 were associated with antibodies to the periodontal pathogen Prevotella intermedia (P = 0.05 and P = 0.001, respectively), but not with antibodies to Porphyromonas gingivalis arginine gingipains. Levels of antibodies to CEP-1, cFIBß, and cVIM correlated with each other, and with smoking and shared epitope risk factors in RA. CONCLUSION: This study identifies 2 groups of ACPA fine specificities associated with different RA risk factors. One is predominantly linked to smoking and shared epitope, and the other links anti-cTNC5 and cCK13-1 to infection with the periodontal pathogen P intermedia.

Periopathogens differ in terms of the susceptibility to toluidine blue O-mediated photodynamic inactivation.

BACKGROUND: The main goal of periodontal therapy is to eliminate the infection spreading in periodontium. Antimicrobial photodynamic therapy may be applied in order to eradicate pathogens remaining in periodontal tissues after conventional mechanical debridement, to improve the treatment results. The aim of this in vitro study was to evaluate the susceptibility of selected key periopathogens to toluidine blue O-mediated photodynamic inactivation and the influence of photosensitizer's concentration and light dose on the effectiveness of this process. METHODS: Following bacterial strains were used in the experiments: Porphyromonas gingivalis ATCC 33277, Aggregatibacter actinomyctemecomitans ATCC 33384, Fusobacterium nucleatum ATCC 10953. Toluidine blue O (TBO) was used in concentration ranging from 0.004 to 0.5mg/mL. Irradiation was performed by a non-laser red light source. RESULTS: Complete eradication of P. gingivalis was obtained upon the application of TBO in the concentration of 0.1mg/mL and the highest light dose. A, actinomycetemcomitans was, in turn, not susceptible to photodynamic inactivation regardless of the dosimetric parameters applied. High viability reductions were also obtained for F. nucleatum, however no complete eradication. The effectiveness of photodynamic inactivation of susceptible periopathogens was dependent on the light dose and photosensitizer's concentration. CONCLUSIONS: Periopathogens differ in terms of their susceptibility to photodynamic inactivation. Antimicrobial PDT may be valuable in the treatment of those cases of periodontal disease, in which P. gingivalis is a dominating pathogen. Microbiological examination prior to clinical application of aPDT may be recommended.

Gingival Fibromatosis with Significant De Novo Formation of Fibrotic Tissue and a High Rate of Recurrence.

BACKGROUND Hereditary gingival fibromatosis is characterized by slowly progressive enlargement of the gingiva that can present as an isolated condition or present as part of various syndromes. CASE REPORT An 11-year-old female reported with a gingival lesion that caused masticatory problems and poor oral hygiene. Periodontal examination revealed a dense tissue covering 30% of her teeth crowns within both jaws. Panoramic x-ray showed a normal bone height and teeth positioning. The patient did not use any medications, but a similar condition was also present in other family members. The patient was diagnosed with hereditary gingival fibromatosis. Surgery was carried out to remove excess of gingival tissue. Post-surgical healing was uneventful, but four weeks after the first surgery, the condition recurred amounting to 45% of the initial tissue volume presenting in the mandible, and 25% in the maxilla. Two months later, no significant growth was noted in the mandible, while in the maxilla, growth increased to 40% of the preoperative state. Analysis by polarized microscope showed a significant increase of thin fibrotic fibrils that contributed 80% of the total pool of collagen fibrils in the patient's gingiva, but only 25% in healthy gingiva. The patient was receiving outpatient care for follow-up every three months and surgical intervention had not been planned as long as her periodontal health was not be compromised.  CONCLUSIONS It is currently not clear whether the extent of the fibrosis had a mechanistic association with the ratio of gingival tissue re-growth in our case study. Further studies are needed to explain this association and improve the management of this condition.