Gingival fibroblast activation by Porphyromonas gingivalis is driven by TLR2 and is independent of the LPS-TLR4 axis.

Gingival fibroblasts (GFs) are abundant structural cells of the periodontium that contribute to the host's innate immunity by producing cytokines and chemokines in response to oral pathogens, such as Porphyromonas gingivalis. Isolated lipopolysaccharide (Pg-LPS) is commonly used to study GF responses to P. gingivalis; however, this approach produced conflicting observations regarding its proinflammatory potential and the engagement of specific Toll-like receptors (TLRs). In this work, we demonstrate that commercially available Pg-LPS preparations are weak activators of GF innate immune responses compared with live P. gingivalis or other relevant virulence factors, such as P. gingivalis fimbriae or LPS from Escherichia coli. GF's nonresponsiveness to Pg-LPS can be only partly attributed to the low expression of TLR4 and its accessory molecules, CD14 and LY36, and is likely caused by the unique structure and composition of the Pg-LPS lipid A. Finally, we combined gene silencing and neutralizing antibody studies to demonstrate that GF response to infection with live P. gingivalis relies predominantly on TLR2. In contrast, the LPS-TLR4 signaling plays a negligible role in inflammatory cytokine production by GFs exposed to this oral pathogen, confirming that Pg-LPS stimulation is not an optimal model for studies of GF responses to P. gingivalis.

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.

Nordic Walking at Maximal Fat Oxidation Intensity Decreases Circulating Asprosin and Visceral Obesity in Women With Metabolic Disorders.

OBJECTIVE: Excess visceral adipose tissue is associated with insulin resistance and other metabolic disorders, including deregulation of adipokine secretion, which may be corrected by aerobic exercise training. Asprosin is a novel adipokine responsible for the regulation of appetite and the release of glucose from the liver, and its levels are pathologically elevated in obesity. The aim of the study was to evaluate the effects of 8-week Nordic walking (NW) training at maximal fat oxidation intensity (FAT max ) on changes in body mass, as well as those in insulin resistance and asprosin levels among young women with visceral obesity and metabolic disorders. MATERIALS AND METHODS: The study was completed by 14 women (30.14 ± 3.63 years) representing low levels of physical activity, visceral obesity (waist circumference 105.50 ± 14.87 cm, BMI 33.85 ± 5.48 kg/m2) and with metabolic disorders, who for 8 weeks (three times a week, 60 min), participated in NW training at the FAT max intensity (61.92 ± 6.71% HR max , 42.33 ± 8.69% VO2max) controlled on the basis of heart rate (114.21 ± 14.10 bpm). RESULTS: After 4 and 8 weeks of NW training, a significant decrease in the concentration of asprosin, waist and hip circumference (HC), waist-to-height ratio and body adiposity index (BAI) (p < 0.05, large effect size) were found. CONCLUSION: The 8-week NW training at an FAT max intensity decreases the concentration of asprosin in the blood as well as visceral obesity in young women with metabolic disorders.

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.

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.

Anaerobic Exercise-Induced Activation of Antioxidant Enzymes in the Blood of Women and Men.

Objective: Physical exercise changes redox balance in the blood. The study aim is to determine gender-related differences in enzymatic antioxidant defense [superoxide dismutase, catalase (CAT), and glutathione peroxidase (GPx)] during the initial period following anaerobic exercise and 24 h after its completion. Methods: Young, non-training participants (10 women and 10 men) performed a single anaerobic exercise, which was a 20-s maximal cycling sprint test. Blood was collected before and after completing the anaerobic exercise, i.e., after 3, 15, 30, and 60 min and after 24 h. Lactate concentration, and the superoxide dismutase, CAT, and GPx activity were determined. The results were adapted to the changes in plasma volume. Results: Anaerobic exercise induced a significant increase in lactate concentration, similar among both sexes. Anaerobic exercise evokes identical changes in the activity of antioxidant enzymes in the blood plasma of women and men, which is dependent on anaerobic capacity. In the early phase of restitution, the activity of antioxidant enzymes decreases; 24 h after anaerobic exercise, GPx activity in the blood plasma of women and men is higher than before the exercise. Conclusion: There are no gender-related differences concerning changes in plasma antioxidant activity after anaerobic exercise. Depending on the antioxidant enzyme, changes of activity differ in time after the end of the anaerobic exercise.

Unchanged Erythrocyte Profile After Exposure to Cryogenic Temperatures in Elder Marathon Runners.

Objective: Endurance runners may experience "sports anemia" resulting from intravascular hemolysis. In addition, aging has negative impact on hematopoiesis and rheological properties of blood, and erythrocyte membranes in older people are more vulnerable to oxidative damage, which together can lead to anemia. Whole-body cryostimulation (WBCST) is increasingly used in the elderly as a method of biological regeneration of athletes or therapy and preventive treatment. That is why the aim of the study was to determine whether repeated WBCST had an effect on the erythrocyte system in master marathon runners, compared to non-training men. Methods: Ten marathon runners (men aged 55.9 ± 5.5 years, training experience 6.71 ± 5.79 years) and 10 non-training (men aged 62.0 ± 5.8 years) were subjected to a series of 24 WBCST (3 min, -130°C) performed every other day. Erythrocyte levels, interleukin-3 (IL-3), erythropoietin (EPO), haptoglobin, bilirubin, and extracellular hemoglobin (HGBecf) concentrations were determined in the blood before and after 12, 24 WBCST, as well as 7 days after their completion. Results: The concentrations of EPO and IL-3 were significantly increased 7 days after the completion of WBCST in both groups (P < 0.05). The erythrocyte content and indicators, the bilirubin, haptoglobin, and HGBecf levels in each group did not change as a result of WBCST. In order to document hemolytic changes and/or factors affecting the severity of erythropoiesis, correlations between growth erythropoietic factors, erythrocyte and hemolytic factors as well as mutual correlations between hemolytic indexes were calculated. There was a positive correlation (P < 0.05) between the EPO and IL-3, bilirubin, mean corpuscular hemoglobin, and red blood cell distribution width - standard deviation. There was also a positive correlation between the concentrations of bilirubin and HGBecf, and a negative correlation between haptoglobin and HGBecf as well as bilirubin concentrations. Conclusion: WBCST treatments, repeated every other day, do not cause hemolytic changes in elder men with high or low physical activity. But also, they are a procedure that does not increase the level of erythrocytes or their hemoglobinization. In athletes, it is not a form of doping. The positive correlation between EPO and bilirubin may be indicative of, for example, the mutual antioxidative effect of these factors.

Acute Anaerobic Exercise Affects the Secretion of Asprosin, Irisin, and Other Cytokines - A Comparison Between Sexes.

Objective: The new adipokine, which is asprosin, affects glucose release from the liver to the blood, and thus, influences exercise metabolism. This is the first study assessing whether single anaerobic exercise affects asprosin secretion in women and men. Methods: 10 men and 10 women (aged 21.64 ± 1.22 and 22.64 ± 1.49, respectively) performed a single 20-s bicycle sprint. Blood samples were collected before exercise and in the 3', 15', 30', and 60' of recovery, and 24 h after competition. Results: Only in women did asprosin (P = 0.001) (15', 30', 60', and 24 h after exercise) and irisin (P < 0.001) (15', 30', and 60') concentrations increase. Leptin, however, decreased (P = 0.001) at 3', 15', and 30' in women. There was an increase in interleukin-6 (P < 0.001) at 3', 15', 30', and 60' of recovery in men, at 15', 30', 60', and 24 h of recovery in women, along with a simultaneous decrease in interleukin-1ß (P < 0.001) at 15', 30', and 60' of recovery in men, and at 15' and 30' of recovery in women (r = -0.35, P < 0.001). There was a positive correlation between asprosin and adiponectin and a negative one between asprosin and leptin. The increase in irisin concentration at 30' of recovery was positively correlated with the increase in asprosin concentration and percentage fat content, while being negatively correlated with total and lean body mass (LBM). Conclusion: The single anaerobic effort induced an increase in asprosin and irisin secretion while reducing leptin secretion in women. Adipocytokine concentration changes are inter-related. Regardless of sex, anaerobic efforts induce anti-inflammatory effects.

Effect of body composition, aerobic performance and physical activity on exercise-induced oxidative stress in healthy subjects.

BACKGROUND: Oxidative stress could be the result of an increase in ATP resynthesis during exercise. The aim of the study was to compare prooxidant-antioxidant balance (PAB) disturbances induced by exercise at maximal intensity in young men with differing body compositions. METHODS: Thirty-nine subjects were selected from 1549 volunteers aged 18-30, based on lean body mass (LBM) and body fat percentage (%BF), and then assigned into one of the following groups: control group (CON), including subjects with average LBM (59.0-64.3 kg) and average %BF (14.0-18.5%); high body fat (HBF) group, including subjects with high %BF (>21.5%) and average LBM; and high lean body mass (HLBM) group, including subjects with high LBM (>66.3 kg) and average %BF. Participants' physical activity was determined. A running test with a gradually increased load was used. Before and 3 minutes after exercise, total oxidative status (TOS) and total antioxidant capacity (TAC) were determined in the plasma, and the Oxidative Stress Index (OSI = TOS/TAC) was calculated. RESULTS: Maximal oxygen uptake (VO2max) was comparable in the HBF and HLBM groups (53.12±1.51 mL/kg and 50.25±1.27 mL/kg, respectively) and significantly lower compared to the CON group (58.23±1.62 mL/kg). The CON, HBF and HLBM groups showed similar significant (P<0.05) increases in TOS levels (36%, 35% and 31%, respectively). Post-exercise TAC increased by 8% in the HBF and HLBM groups (P<0.05), compared to the 3% increase in the CON group (P>0.05). There was significant negative correlation between OSI, measured before and after exercise, and participants' physical activity. There was no correlation between OSI and VO2max, BM, LBM, %BF and BMI. CONCLUSIONS: Exercise at maximal intensity causes a similar increase in TOS and in TAC in subjects with increased %BF and elevated content of LBM and regardless of body composition, the ratios of TOS/TAC concentrations before and after maximal-intensity exercise, have lower values in people with higher physical activity levels and are not dependent on aerobic performance (VO2max).

Impact of single anaerobic exercise on delayed activation of endothelial xanthine oxidase in men and women.

OBJECTIVES: The aim of the study was to evaluate the activity of xanthine oxidase (XO) in the blood of men and women during the first hour following a single anaerobic exercise (AN-EX), and after 24 hours of recovery, and to determine whether the changes in XO activity in the blood after AN-EX are dependent on anaerobic performance. METHODS: Ten men and ten women performed a single AN-EX. Blood was collected before and five times after completion of the AN-EX. The activity of XO was determined. RESULTS: In both groups, a significant (P < 0.05) increase in blood XO activity was found only 24 hours after the AN-EX. The increased activity of XO in men was significantly lower than in women (P < 0.05). Negative correlations were found between the increase in XO activity in the blood plasma 24 hours after the AN-EX and anaerobic power, the total work performed during the AN-EX and the power decrease. DISCUSSION: In the first hour after the single AN-EX, XO activity in the blood of women and men did not change, but after 24 hours of recovery, it was significantly higher compared to baseline levels in both sexes. Single AN-EX causes a smaller increase in XO activity in people with higher anaerobic performance.

Changes in Non-Enzymatic Antioxidants in the Blood Following Anaerobic Exercise in Men and Women.

PURPOSE: The aim of this study was to compare changes in total oxidative status (TOS), total antioxidative capacity (TAC) and the concentration of VitA, VitE, VitC, uric acid (UA), reduced (GSH) and oxidized glutathione (GSSG) in blood within 24 hours following anaerobic exercise (AnEx) among men and women. METHODS: 10 women and 10 men performed a 20-second bicycle sprint (AnEx). Concentrations of oxidative stress indicators were measured before AnEx and 3, 15 and 30 minutes and 1 hour afterwards. UA, GSH and GSSH were also measured 24 hours after AnEx. Lactate and H+ concentrations were measured before and 3 minutes after AnEx. RESULTS: The increase in lactate and H+ concentrations following AnEx was similar in both sexes. Changes in the concentrations of all oxidative stress indicators were significant and did not differ between men and women. In both sexes, TOS, TAC, TOS/TAC and VitA and VitE concentrations were the highest 3 minutes, VitC concentration was the highest 30 minutes, and UA concentration was the highest 1 hour after AnEx. GSH concentration was significantly lower than the initial concentration from 15 minutes to 24 hour after AnEx. GSSG concentration was significantly higher, while the GSH/GSSG ratio was significantly lower than the initial values 1 hour and 24 hour after AnEx. CONCLUSIONS: With similar changes in lactate and H+ concentrations, AnEx induces the same changes in TAC, TOS, TOS/TAC and non-enzymatic antioxidants of low molecular weight in men and women. Oxidative stress lasted at least 24 hours after AnEx.

[Erythema exudativum multiforme--case report]. / Rumien wysiekowy wielopostaciowy--opis przypadku.

Erythema exudativum multiforme (EM) is an acute mucocutaneous disease. Etiology of this disease is not clear. 50% - 60% cases of EM is induced by HSV infection (herpes simplex virus 1, herpes simplex virus 2). EM lesions may appear as red macules, erosions, ulcerations and hemorrhagic crusts. The study describes the case of 22 - year - old male who had characteristic lesions for EM after HSV infection.

Influence of diet on oral health in young adults--pilot study.

INTRODUCTION: Diet is a factor that can modify, among others, the course of caries, dental erosive lesions and periodontal disease. It is important to know the influence of diet on the clinical oral condition in young adults. MATERIAL AND METHODS: 20 women and 20 men aged 19-21 participated in the study. During a clinical examination of the patients the following clinical indices were estimated: Ap- proximal Plaque Index (API), Bleeding on Probing (BOP) and DMFT indices. The frequency and quality of food products consumed were noted in a special questionnaire. Results with p ≤ 0.05 were considered statistically significant. RESULTS: The average API value for the female participants was 77.37%, BOP--23.84% and DMFT--9.9. The t-test results for independent samples showed that women who consumed meat and meat products two times a week or more, had statistically higher API (p = 0.01) and BOP (p = 0.05) values, than the mean value for these indexes. Women who consumed grain products and grain-based products two times a week or more, had higher API values compared to the mean value for this index (p = 0.02). The average API value for the male participants was 34.25%, BOP--10.15% and DMFT--9.05, respectively. Men who consumed milk and dairy products three times a week or less had statistically lower API (p = 0.05) and BOP values (p = 0.02), relative to the mean index values. CONCLUSIONS: The chemical composition and texture of the food intake can affect oral health. Besides instruction in oral hygiene, doctors and dentists should also assess the eating habits of their patients.

The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis.

Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires heme from host hemoproteins using the HmuY hemophore. The aim of this study was to develop a specific P. gingivalis marker based on a hmuY gene sequence. Subgingival samples were collected from 66 patients with chronic periodontitis and 40 healthy subjects and the entire hmuY gene was analyzed in positive samples. Phylogenetic analyses demonstrated that both the amino acid sequence of the HmuY protein and the nucleotide sequence of the hmuY gene are unique among P. gingivalis strains/isolates and show low identity to sequences found in other species (below 50 and 56%, respectively). In agreement with these findings, a set of hmuY gene-based primers and standard/real-time PCR with SYBR Green chemistry allowed us to specifically detect P. gingivalis in patients with chronic periodontitis (77.3%) and healthy subjects (20%), the latter possessing lower number of P. gingivalis cells and total bacterial cells. Isolates from healthy subjects possess the hmuY gene-based nucleotide sequence pattern occurring in W83/W50/A7436 (n = 4), 381/ATCC 33277 (n = 3) or TDC60 (n = 1) strains, whereas those from patients typically have TDC60 (n = 21), W83/W50/A7436 (n = 17) and 381/ATCC 33277 (n = 13) strains. We observed a significant correlation between periodontal index of risk of infectiousness (PIRI) and the presence/absence of P. gingivalis (regardless of the hmuY gene-based sequence pattern of the isolate identified [r = 0.43; P = 0.0002] and considering particular isolate pattern [r = 0.38; P = 0.0012]). In conclusion, we demonstrated that the hmuY gene sequence or its fragments may be used as one of the molecular markers of P. gingivalis.

[Nutrition and periodontal diseases]. / Odzywianie a choroby przyzebia.

The aim of this paper is to review the available literature pertaining the influence of specific nutritional elements (e.g. vitamin C, vitamin B-complex and dietary calcium) on periodontal status. Effect of food intake on the oral cavity is complex. Proper nutrition, both in terms of quantity and quality determines health of oral cavity and entire body.

[Local complications of lip and tongue piercing -- report of the cases]. / Miejscowe powiklania kolczykowania wargi i jezyka -- opis przypadków.

Piercing, which is the form of puncturing parts of the human body and creating openings where jewelry can be worn, is together with tattoos, body drawings or make-up, one of the oldest way of decorating of the human body. Piercing can be performed in each part of the body. In the recent years face piercing (nose, eyebrows, lip region) and in the oral cavity (tongue, lips, cheek, mentolabial sulcus, labial frenulum) is becoming particularly popular. The procedure of piercing itself may potentially lead to serious systemic and local complications. The aim of the study was to present three clinical cases of the patients who referred to the Dental University Clinic in Krakow for the treatment of lesions on the oral mucosa after piercing. Clinicians who examine patients with such body decorations should pay particular attention to the sites which can be injured by the jewelry. Medical staff should also make patients realize the risk of general complications after piercing. This will allow on the early removal of the jewelry and prevention of possible complications.

[Diabetes mellitus and periodontal disease]. / Zapalenie przyzebia w przebiegu cukrzycy.

Scientific studies confirm correlation between periodontitis and systemic diseases such as: arteriosclerosis, diabetes, heart diseases, stroke, diseases of the respiratory system, kidney diseases, osteoporosis, rheumatoid arthritis, premature birth and low birth weight. The interaction between periodontitis and diabetes mellitus is described, based on the literature.