Simulated in vitro hypoxic conditions from psoriatic arthritis cartilage change plasminogen activating system urokinase and serpine functionality. Reversal of antiapoptotic protection suggests common homeostatic buffering.

Introduction: Rheumatoid and psoriatic arthritis are both characterised by synovial destruction associated with a higher turnover of the extracellular matrix. In both conditions, inflammatory processes create hypoxic environments which destabilise members of the plasminogen activating system. Aim: Comparing the effect of bioactive concentrations of urokinase (uPA) and serpine (PAI-1) on cellular survival of human fibroblast-like-synoviocytes (HFLS) in rich and hypoxic growth media. Material and methods: Monocultures of HFLS were exposed to bioactive uPA and PAI-1 concentrations in both media conditions for 24, 48 and 72 h. Cellular survival was evaluated with a cell viability assay by spectrum absorbance of formazan reduced WST-8. Results: PAI-1 at 0.1 and 1 µg/ml was found to stimulate cell viability under hypoxic stress at 48 and 72 h of incubation, with the effect increasing from 48 to 72 h. uPA increased cell viability in rich medium at 48 and 72 h of incubation between 5 and 40 ng/l, but was found to reduce viability at 80 ng/l at 24 and 48 h. PAI-1 increased cell viability in the hypoxic stress model, while high concentrations of uPA decreased cell viability in rich medium. Conclusions: The alternative modes of function at extreme concentrations provide a novel description of PAI-1 and uPA activity based on their colocalization and mutual buffering capacity, helping to place these molecules more accurately in the context of arthritic synovial deterioration.

Cytotoxic Potential of Denture Adhesives on Human Fibroblasts-In Vitro Study.

(1) In recent years, there has been a significant increase in the availability of denture adhesives for stabilizing removable dentures. The aim of the present study was to assess the cytotoxicity of three denture adhesives on human fibroblasts. (2) Methods: Three denture adhesives were analyzed. Fibroblast cultures were established for the study and control groups in order to assess the incidence of necrosis and to evaluate the microscopic intracellular alterations induced. Following incubation with (study groups) or without adhesives (control group), trypan blue dye exclusion assay was used to determine the number of viable and/or dead cells. Microscopic specimens were stained with haematoxylin and eosin, scanned, digitally processed and then analyzed by a histopathologist. (3) Results: All three denture adhesives analyzed demonstrated various toxic effects in vitro on human fibroblast: quantitative evaluation-45.87-61.13% reduction of cell viability (p = 0.0001) and slight to moderate cytotoxicity in qualitative evaluation. (4) Conclusions: Denture adhesive creams demonstrated a toxic effect on human fibroblasts in vitro in quantitative and qualitative evaluation. In vivo observations are needed to find out if denture adhesives present a cytotoxic effect in patients.

Influence of Chlorhexidine and Cetylpyridine on Periodontal Status and Indicators of Oxidative Stress in Patients with Type 1 Diabetes.

OBJECTIVE: One of the treatment goals in type 1 diabetes and periodontitis is to address chronic inflammation to prevent the development of neurovascular complications. The aim of this study was to assess the local anti-inflammatory effects of chlorhexidine digluconate and cetylpyridine chloride on periodontal status and indicators of oxidative stress in saliva in patients with type 1 diabetes. MATERIALS AND METHODS: A total of 42 subjects aged 27 (interquartile range, IQR 22-35) years, with type 1 diabetes for a duration of 12 (IQR 9-18) years, and glycated hemoglobin 8.05 (IQR 7.1-9.4)% were included. Patients were examined twice-initially, and after 14 days of using toothpaste with chlorhexidine and cetylpyridine. Clinical examination of gingival tissues was performed. Certain oxidative stress markers (TP, TEAC, TBARS, AOPP) were measured in the saliva samples. RESULTS: There were significant changes in clinical indicators of periodontal status before and after the application of the toothpaste (API before 0.35 (0.24-0.65) vs. API after 0.265 (0.18-0.39), p = 0.03; SBI before 0.07 (0.04-0.15) vs. SBI after 0.035 (0-0.06), p = 0.002; GI before 0.88 (0.46-1) vs. GI after 0.67 (0.25-1), p = 0.0008). The concentration of saliva TBARS decreased (p = 0.00005) and TEAC increased (p = 0.09). CONCLUSION: Proper oral hygiene supported by antibacterial chemicals may improve the periodontal status and reduce inflammation.

A comprehensive review of the influence of Epigallocatechin gallate on Sjögren's syndrome associated molecular regulators of exocytosis (Review).

Sjögren's syndrome (SS) is an autoimmune disorder that affects the salivary glands, leading to reduced secretory functions and oral and ocular dryness. The salivary glands are composed of acinar cells that are responsible for the secretion and production of secretory granules, which contain salivary components, such as amylase, mucins and immunoglobulins. This secretion process involves secretory vesicle trafficking, docking, priming and membrane fusion. A failure during any of the steps in exocytosis in the salivary glands results in the altered secretion of saliva. Soluble N-ethylmaleimide-sensitive-factor attachment protein receptors, actin, tight junctions and aquaporin 5 all serve an important role in the trafficking regulation of secretory vesicles in the secretion of saliva via exocytosis. Alterations in the expression and distribution of these selected proteins leads to salivary gland dysfunction, including SS. Several studies have demonstrated that green tea polyphenols, most notably Epigallocatechin gallate (EGCG), possess both anti-inflammatory and anti-apoptotic properties in normal human cells. Molecular, cellular and animal studies have indicated that EGCG can provide protective effects against autoimmune and inflammatory reactions in salivary glands in diseases such as SS. The aim of the present article is to provide a comprehensive and up-to-date review on the possible therapeutic interactions between EGCG and the selected molecular mechanisms associated with SS.

Potential usefulness of enamel matrix derivative in skin and mucosal injury treatment.

Enamel matrix proteins (EMP) are secreted by ameloblasts during odontogenesis. The main component of enamel protein extract is amelogenin. The extracts also contain proteins with bioactive properties similar to bone morphogenic proteins and transforming growth factor ß1. Research on animal models indicates that EMP improve healing of oral mucosa wounds by stimulating the production of collagen fibers and blood vessels in the connective tissue. Success in the treatment of oral wounds prompted interest in possible applications of amelogenins in the repair of damaged skin due to similarities in histological structure between skin and mucosa.

Effect of injectable equine collagen type I on metabolic activity and apoptosis of gingival fibroblasts.

Collagen as a biomaterial is widely used for tissue regeneration due to various advantages including its biodegradation, biocompatibility, and low allergenicity. Along with aesthetic medicine development, collagen is also used in the injectable form as a tissue biostimulator. The area of our study was collagen's impact on fibroblast activity and apoptosis. The research showed that atelocollagen decreases metabolic activity of fibroblasts, but also showed an increasing number of living cells after 48 h and 72 h incubation under the influence of collagen.

Adipose-PAS interactions in the context of its localised bio-engineering potential (Review).

Adipocytes are a known source of stem cells. They are easy to harvest, and are a suitable candidate for autogenous grafts. Adipose derived stem cells (ADSCs) have multiple target tissues which they can differentiate into, including bone and cartilage. In adipose tissue, ADSCs are able to differentiate, as well as providing energy and a supply of cytokines/hormones to manage the hypoxic and lipid/hormone saturated adipose environment. The plasminogen activation system (PAS) controls the majority of proteolytic activities in both adipose and wound healing environments, allowing for rapid cellular migration and tissue remodelling. While the primary activation pathway for PAS occurs through the urokinase plasminogen activator (uPA), which is highly expressed by endothelial cells, its function is not limited to enabling revascularisation. Proteolytic activity is dependent on protease activation, localisation, recycling mechanisms and substrate availability. uPA and uPA activated plasminogen allows pluripotent cells to arrive to new local environments and fulfil the niche demands. However, overstimulation, the acquisition of a migratory phenotype and constant protein turnover can be unconducive to the formation of structured hard and soft tissues. To maintain a suitable healing pattern, the proteolytic activity stimulated by uPA is modulated by plasminogen activator inhibitor 1. Depending on the physiological settings, different parts of the remodelling mechanism are activated with varying results. Utilising the differences within each microenvironment to recreate a desired niche is a valid therapeutic bio-engineering approach. By controlling the rate of protein turnover combined with a receptive stem cell lineage, such as ADSC, a novel avenue on the therapeutic opportunities may be identified, which can overcome limitations, such as scarcity of stem cells, low angiogenic potential or poor host tissue adaptation.

Diagnosis of Papillon-Lefèvre syndrome: review of the literature and a case report.

Papillon-Lefëvre syndrome (PLS), classified as ectodermal dysplasia, is an autosomal recessive condition related to the cathepsin C (CTSC) gene mutation. The first clinical symptoms, occurring most commonly between the ages of 1 and 4, are palmoplantar hyperkeratosis and also periodontitis resulting in the loss of most or all teeth in the same sequence in which they erupted. Most often the redness of palms and soles precede the occurrence of keratoderma. Moreover, excessive sweating, moderate mental retardation, the tendency to purulent skin and internal organs infection may occur. Lack of cathepsin seems to have a crucial role in the intensity of symptoms. In most of the patients, there can be observed impairment of phagocytosis and chemotaxis of neutrophils, granulocytes, leukocytes and cytotoxic lesion of fibroblasts and macrophages. Also, functional impairment of lymphocytes, neutrophils, and monocytes is observed. The study, using flow cytometry, showed a decreased percentage of T cells CD8+ and increased CD4:CD8 ratio.

Salivary Cytokine Profile as a Possible Predictor of Autism Spectrum Disorder.

Autism spectrum disorder (ASD) is characterized by neurodevelopmental disorders and alterations in immune function and cytokine levels. The aim of this study is to determine the salivary levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor α (TNFα), monocyte chemoattractant protein-1 (MCP-1), Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES), and Eotaxin in children with ASD and in healthy controlsto assess their predictive potential. We explored correlations between the cytokine levels and the neurodevelopmental disorders related to ASD. The study comprised 19 children with ASD and 19 typically developing (TD) ones. We analyzed salivary levels of IL-1ß, IL-6, IL-8, TNFα, MCP-1, RANTES, and eotaxin on Luminex with custom-designed 7-plex kits. The level of RANTES in ASD children was significantly lower than those of TD. In TDs, the salivary levels of IL-1ß, MCP-1, and TNFα correlated positively with age. In ASD, the cytokine levels did not correlate with age. There were statistically significant differences between the RANTES level and aggression and gait disturbances, between IL-8 level and fixations/stimulations, and between IL-1ß level and no active speech. The levels of the cytokine detected can manifest both systemic and local changes related to ASD. The cytokine pattern cannot be used as a sole ASD predictor, but the salivary levels may be helpful in categorizing the ASD subtype.

The relationship between periodontal status and rheumatoid arthritis - systematic review.

There have been numerous publications investigating the relationship between periodontitis (PD) and rheumatoid arthritis (RA) so far. This publication presents the common risk factors for the development of PD and RA. The major impact of the pathological bacterial factor and cigarette smoking with chronic inflammation playing the key role in both diseases has been confirmed by numerous studies in various populations over the years. More research focuses nowadays also on the role of improper diet and obesity. Pathophysiological pathways, such as increased concentration of proinflammatory cytokines, indirectly affecting the cardiovascular complications and coagulation disorders, which has an impact on function disorders of tissue metalloproteinase inhibitors and the plasminogen activation system, were also researched. This systematic review of current literature has shown numerous discrepancies in previous analyses and the need for further detailed research on the relationship between periodontal status and RA.

Atelocollagen Application in Human Periodontal Tissue Treatment-A Pilot Study.

BACKGROUND: The aim of this study is the clinical observation of gingival tissue condition after atelocollagen injection. METHODS: In 18 patients, 97 gingival class I Miller recessions were divided according to recession height, gingival papillae loss and thickness of gingivae. Atelocollagen (Linerase, 100 mg) was injected into keratinized gingivae twice or thrice, at two-week intervals. RESULTS: Statistically significant changes in gingival recession, amount of gingival papillae loss and thickness of gingiva were observed, after both two and three collagen injections. Although the degree (height) of recession decreased and gingival tissue thickness increased with every injection; there was no difference in gingival papillae loss between second and third collagen injections. CONCLUSIONS: The injectable form of atelocollagen is a promising material for gingival soft tissue regeneration and stimulation and allows for reduction in the number of procedures and support in a variety of surgical scenarios. This is a pilot study that clinically measures the impact of injected atelocollagen on periodontal tissue biotype, including the thickness of gingivae and gingival papillae regeneration.

Influence of Herbal Medicines on HMGB1 Release, SARS-CoV-2 Viral Attachment, Acute Respiratory Failure, and Sepsis. A Literature Review.

By attaching to the angiotensin converting enzyme 2 (ACE2) protein on lung and intestinal cells, Sudden Acute Respiratory Syndrome (SARS-CoV-2) can cause respiratory and homeostatic difficulties leading to sepsis. The progression from acute respiratory failure to sepsis has been correlated with the release of high-mobility group box 1 protein (HMGB1). Lack of effective conventional treatment of this septic state has spiked an interest in alternative medicine. This review of herbal extracts has identified multiple candidates which can target the release of HMGB1 and potentially reduce mortality by preventing progression from respiratory distress to sepsis. Some of the identified mixtures have also been shown to interfere with viral attachment. Due to the wide variability in chemical superstructure of the components of assorted herbal extracts, common motifs have been identified. Looking at the most active compounds in each extract it becomes evident that as a group, phenolic compounds have a broad enzyme inhibiting function. They have been shown to act against the priming of SARS-CoV-2 attachment proteins by host and viral enzymes, and the release of HMGB1 by host immune cells. An argument for the value in a nonspecific inhibitory action has been drawn. Hopefully these findings can drive future drug development and clinical procedures.

Proteolysis is the most fundamental property of malignancy and its inhibition may be used therapeutically (Review).

The mortality rates of cancer patients decreased by ~1.5% per year between 2001 and 2015, although the decrease depends on patient sex, ethnic group and type of malignancy. Cancer remains a significant global health problem, requiring a search for novel treatments. The most common property of malignant tumors is their capacity to invade adjacent tissue and to metastasize, and this cancer aggressiveness is contingent on overexpression of proteolytic enzymes. The components of the plasminogen activation system (PAS) and the metalloproteinase family [mainly matrix metalloproteinases (MMPs)] are overexpressed in malignant tumors, driving the local invasion, metastasis and angiogenesis. This is the case for numerous types of cancer, such as breast, colon, prostate and oral carcinoma, among others. Present chemotherapeutics agents typically attack all dividing cells; however, for future therapeutic agents to be clinically successful, they need to be highly selective for a specific protein(s) and act on the cancerous tissues without adverse systemic effects. Inhibition of proteolysis in cancerous tissue has the ability to attenuate tumor invasion, angiogenesis and migration. For that purpose, inhibiting both PAS and MMPs may be another approach, since the two groups of enzymes are overexpressed in cancer. In the present review, the roles and new findings on PAS and MMP families in cancer formation, growth and possible treatments are discussed.

Benign tumours and tumour-like lesions in the oral cavity: a retrospective analysis.

INTRODUCTION: Oral lesions are divided into non-neoplastic lesions, potentially malignant lesions and neoplastic lesions. More clinical data are needed to determine their helpful clinical pattern. AIM: To present the epidemiological, clinical and histopathological characteristics of the oral lesions. MATERIAL AND METHODS: The retrospective study group comprised records of 208 patients which were reviewed according to selected epidemiological and clinical features. All the biopsy specimens were classified into: reactive lesions, precancerous lesions/potentially malignant lesions, salivary gland pathologies, benign and malignant tumours. RESULTS: The lower lip was the most common site involved followed by buccal and vestibular mucosa. The most frequent diagnoses were fibroma, mucocele and papilloma. The predominant pathomorphological forms were nodule and bulla. The most frequent salivary gland pathology was mucocele. Fibroma was the most frequent pathomorphological diagnosis, followed by mucocele and reactive lesions such as irritation fibroma (IF) and granuloma. CONCLUSIONS: In cases of oral mucosal lesions, we propose the following algorithm: the exclusion of all odontogenic and iatrogenic causes; the detection and elimination of harmful habits, parafunctions and irritants from the oral cavity especially from the vestibule of the oral cavity and from the lips; all surgical treatment should be performed only after the proper detection and elimination of causative factors to decrease the risk of recurrence; excisional biopsy or in more diffuse lesions incisional biopsy is recommended to confirm clinical diagnosis; and consideration of other factors that can modify the clinical pattern of oral lesions, such as oral hygiene, systemic diseases, and drugs.

Ethanol influence on gingival fibroblasts - a real-time in vitro study.

INTRODUCTION: Alcohol consumption is the world's third largest risk factor for disease and disability. According to the WHO report from 2011: 71% of urban respondents ty and 77% of rural respondents admit to alcohol consumption]. Lower socio-economic status and educational levels result in a greater risk of alcohol-related injury, disease and death. Alcohol is a common component of many medicines, as well as an ingredient in many oral hygiene home products. Mouthwashes containing alcohol are considered to inhibit wound healing in the oral cavity. Due to the fact that many different results are described for different concentrations of alcohol at different times, an attemptwas made to visualise the direct impact of 7.2% and 22% alcohol on human gingival fibroblasts. MATERIAL AND METHODS: PANsystem 2000 was used for visualisation of the reaction of human gingival fibroblasts isolated from gingiva on ethanol in 2 different concentrations. PANsys 3000 is a multi-system fully-automated cell culture device used for in vitro culture and to study a variety of cell lines under conditions similar to in vivo. Observations were carried out for 48 hours since alcohol addition. Pictures were taken in a continuous process at 5 minute intervalds and combined into a film. RESULTS: Both contamination of 7.2% and 22% ethyl alcohol negatively affected morphology and cell proliferation. Addition of ethanol at a concentration 7.2% enabled cells to regain their ability to divide and recover normal morphology after 10 hours; changes caused by 22% ethanol, however, were irreversible. CONCLUSIONS: The obtained results suggest that daily usage of 7.2% alcohol contained in mouthwashes is non-toxic for gingival fibroblasts, and could be recommended after periodontal surgery.

Consumption of cranberry functional beverage reduces gingival index and plaque index in patients with gingivitis.

Periodontal disease is highly prevalent worldwide, and consumption of certain foods, such as fruits, seem to improve the effectiveness of periodontal therapy (PT) due to their antiadhesive, immunomodulatory, and antioxidative properties. We hypothesized that the cranberry functional beverage (CFB) consumed for eight weeks improves gingival inflammation indices via inhibition of dental plaque, and alterations in antioxidant status, and systemic inflammation in patients with gingivitis. In this two-arm randomized controlled study, fifty participants were divided into an experimental group (CFB), administered daily with 750 ml CFB, or a control group administered the same amount of water. All patients underwent nonsurgical PT prior to the intervention. Gingival (GI) and bleeding on probing (BoP) indices of inflammation, plaque (PI) and approximal plaque (API) indices of dental plaque deposition, saliva and serum total antioxidant status (TAS), serum malonylodialdehyde level (MDA), and interleukin 1-beta level (IL-1beta) were measured pre- and postintervention. A risk of caries development was determined by Streptococcus mutans (SM) and Lactobacillus spp. (LAB) counts in supragingival dental plaque. Changes in GI and PI but not BoP and API were significantly more pronounced in the CFB group compared to the control group. Serum or saliva TAS, IL-1beta, and MDA did not differ between groups. The number of SM reduced in CFB, but not in the control group. We demonstrated that the consumption of CFB improves gingival and plaque indices without posing a risk of caries development. Thus CFB can be recommended as a safe adjunct for nonsurgical PT in patients with gingivitis.

Tobacco smoking decreases clinical symptoms of gingivitis in patients with type 1 diabetes-a cross-sectional study.

OBJECTIVE: Periodontal disease may develop on the background of microvascular complications of diabetes. However, some modifying factors, such as tobacco smoking, should be taken into consideration when assessing risk of development of chronic complications. The aim of the study was the clinical assessment of the periodontal status in patients with type 1 diabetes according to tobacco smoking. SUBJECTS AND METHODS: A total of 362 subjects aged 29 (IQR 22-35) years, type 1 diabetes duration 12 (8-18) years, hemoglobin A1c, HbA1c 8.0 (7.2-9.1)% were included. We used Gingival Index, Approximal Plaque Index, and Sulcus Bleeding Index to assess periodontal health. Patients were divided into two subgroups according to current cigarette smoking. RESULTS: No differences in age, diabetes duration, and chronic complications were found between subgroups. A better metabolic control of diabetes expressed by lower HbA1c (p = 0.00001) and triglyceride levels (p = 0.004) was shown in nonsmokers. Smokers presented significantly lower gingival bleeding, p = 0.009. HbA1c correlated with API in study group (Rs = 0.16; p = 0.002) and in nonsmokers subgroup (Rs = 0.2;p = 0.004), however, not in smoker's subgroup. In multivariable regression analysis, smoking cigarettes (ß = -0.26; p = 0.0002), hs-CRP (ß = 0.15; p = 0.03) and age (ß = -0.19; p = 0.007) occurred to be independent predictors of SBI after adjustment for confounding variables (R2  = 0.13; p = 0.001). CONCLUSIONS: Patients with type 1 diabetes smoking cigarettes presented lower gingival sulcus bleeding and worse metabolic control of diabetes than nonsmoking patients.

Type 1 Diabetes and Periodontal Health.

It is well established that hyperglycemia affects periodontal outcomes. A body of evidence, predominantly over the past 20 years supports significant independent associations between periodontal disease and glycemic control or complications of diabetes. Association between periodontal tissue and hyperglycemia is possible through altered cellular immunity, increased proliferation of bacteria, microangiopathy, and formation of the advanced glycation end products. However, most studies focus solely on patients with type 2 diabetes or diabetes in general. There is still the paucity of data concerning patients with type 1 diabetes (T1D). Here, the authors consider the possible mechanisms linking periodontal disease with diabetes, focusing mainly on T1D and discuss possible diagnostic and therapeutic approaches.

Can EGCG Alleviate Symptoms of Down Syndrome by Altering Proteolytic Activity?

Down syndrome (DS), also known as "trisomy 21", is a genetic disorder caused by the presence of all or part of a third copy of chromosome 21. Silencing these extra genes is beyond existing technology and seems to be impractical. A number of pharmacologic options have been proposed to change the quality of life and lifespan of individuals with DS. It was reported that treatment with epigallocatechin gallate (EGCG) improves cognitive performance in animal models and in humans, suggesting that EGCG may alleviate symptoms of DS. Traditionally, EGCG has been associated with the ability to reduce dual specificity tyrosine phosphorylation regulated kinase 1A activity, which is overexpressed in trisomy 21. Based on the data available in the literature, we propose an additional way in which EGCG might affect trisomy 21-namely by modifying the proteolytic activity of the enzymes involved. It is known that, in Down syndrome, the nerve growth factor (NGF) metabolic pathway is altered: first by downregulating tissue plasminogen activator (tPA) that activates plasminogen to plasmin, an enzyme converting proNGF to mature NGF; secondly, overexpression of metalloproteinase 9 (MMP-9) further degrades NGF, lowering the amount of mature NGF. EGCG inhibits MMP-9, thus protecting NGF. Urokinase (uPA) and tPA are activators of plasminogen, and uPA is inhibited by EGCG, but regardless of their structural similarity tPA is not inhibited. In this review, we describe mechanisms of proteolytic enzymes (MMP-9 and plasminogen activation system), their role in Down syndrome, their inhibition by EGCG, possible degradation of this polyphenol and the ability of EGCG and its degradation products to cross the blood-brain barrier. We conclude that known data accumulated so far provide promising evidence of MMP-9 inhibition by EGCG in the brain, which could slow down the abnormal degradation of NGF.