Gingival Fluid Levels Research Articles

MMP-8 in Periodontal Sites of Postpartum and without-Any-Pregnancy Women.

The hypothesis that physiological changes in women can affect periodontal tissues is the subject of this study, and inflammatory markers such as matrix metalloproteinase-8 can measure susceptibility to inflammation. The study aimed to analyze MMP-8 levels in periodontal sites of postpartum women and women without a history of pregnancy, comparing health parameters and periodontal disease. This is a case-control study with 40 participants, 20 cases (women in the postpartum period) and 20 controls (women without any pregnancy), who underwent clinical periodontal examination and the collection of crevicular gingival fluid. The ELISA test was used to detect MMP-8 levels. Postpartum women had worse periodontal parameters, such as bleeding index on probing, number of sites with CAL ≥ 3, and fewer teeth present. In the group of women without a history of pregnancy, a significantly lower MMP-8 level was observed in healthy sites and a higher one was observed in periodontal pockets (p < 0.01). In contrast, in postpartum women, MMP-8 levels were elevated in both healthy sites and periodontal pockets (p > 0.01). The MMP-8 levels in gingival fluid appear to be related to periodontal clinical parameters and may be a possible marker of enzymatic changes involved in periodontal tissue destruction in postpartum women.

Non-Surgical Periodontal Therapy Could Improve the Periodontal Inflammatory Status in Patients with Periodontitis and Chronic Hepatitis C.

Non-surgical periodontal therapy (NSPT) is the first essential step for the management of any periodontitis patient. This study aims to evaluate the impact of NSPT on pro-inflammatory mediators’ regulation and on clinical parameters in periodontitis patients who suffer from chronic hepatitis C. At baseline, selected patients were clinically evaluated for their periodontal status. A subsequent quantitative assessment of C-reactive protein and pentraxin-3 in samples of gingival fluid was performed by Enzyme-Linked Immunosorbent Assay (ELISA). Afterwards, NSPT was performed. Three months after NSPT, the clinical and ELISA assessments were repeated. The results show an improvement of the clinical parameters in periodontitis patients at the three-month recall. In chronic hepatitis C patients with periodontitis, the gingival fluid levels of pro-inflammatory markers reduced significantly. The targeted markers also expressed significant correlations with the clinical parameters used for the assessment of periodontitis’ severity. The results suggest that, while chronic hepatitis C patients exhibited a more negative periodontal status at baseline as compared to non-hepatitis ones, NSPT is effective in decreasing the local periodontal inflammatory reaction and in proving the periodontal status of this type of patients. Given the limitation of the study, periodontal screening and NSPT should be included in the integrated therapeutical approach of chronic hepatitis C patients, for its impact on the local inflammatory response.

Influence of rs6667202 SNP on Interleukin-10 levels in the gingival fluid of patients with periodontitis grade C

Grade C periodontitis in youngers is characterized by a severe form of periodontitis, and IL10 rs6667202 single nucleotide polymorphism (SNP) has been described as an important feature in this disease etiology. Aim: This study aimed to evaluate, in vivo, the functionality of IL10 rs6667202 SNP on IL-10 gingival fluid levels. Methods: Thirty patients with Perio4C were selected, 15 with the IL10 AA genotype (rs6667202) and 15 with AC/CC genotypes. The gingival fluid was collected from two sites with probing depth ≥ 7 mm and bleeding on probing, and two healthy sites. The IL-10 concentration was determined by Luminex/MAGpix platform. Results: In deep pockets, the IL10 AA genotype presented a lower concentration of IL-10 when compared with AC or CC genotypes (p&lt;0.05). In shallow pockets, no difference between groups was seen (p&gt;0.05). Conclusion: IL10 rs6667202 SNP decreases the production of IL-10 in crevicular fluid, potentially affecting this disease progression.

Diagnostic information content of indicators of cytokine profile of gingival fluid in pregnant women with iron deficiency anemia

Relevance. Modern literature and clinical dental practice do not provide enough information about the value of local immunity parameters for early diagnosis of inflammatory periodontal diseases in pregnant women with iron deficiency anemia. Conventional anti-inflammatory medications used to treat inflammatory periodontal disease are frequently contraindicated during pregnancy. It is important to make a reasonable choice of safe medications, prevention methods, and treatment at early stages of inflammatory periodontal diseases in pregnant women with iron deficiency anemia. The aim of the present study is to determine informative cytokine levels in gingival fluid for early diagnosis of inflammatory periodontal disease and evaluation of effectiveness of “Lysobact” medication in pregnant women with iron deficiency anemia (IDA).Materials and methods. Cytokine levels (IL-8, IL-10; IgA, IgG) in gingival crevicular fluid were investigated in 46 pregnant women aged 18-35: among them 16 (n = 16) had mild IDA (D50), 15 (n = 15) didn't have IDA, 15 (n = 15) were non-pregnant controls. «Lysobact» (Reg. #ПN0179/01-2002) was used in pregnant women with IDA; conventional antiinflammatory therapy was used to treat pregnant women without IDA, before and after drug therapy (8-10 days) according to clinical examinations.Results. There was determined diagnostic value of cytokine levels (IgA, IgG; IL-8, IL-10) that are of great importance for early diagnosis, evaluation of the course of the disease and effectiveness of “Lysobact” immunomodulating drug, and that significantly increase in pregnant women with IDA (IL-10 from 0.010 [1.790] to 3.050 [8.550] ng/ml) versus pregnant women without IDA (from 0.001 [1.259] to 2.900 [18.640] ng/ml) in comparison with controls (4.5 [1.13] ng/ml).Conclusion. Clinical laboratory results in pregnant women with inflammatory periodontal disease allowed to determine diagnostic value of gingival crevicular fluid parameters (sIgA, IgG; IL-8, IL-10) that are related to clinical periodontal index levels, and are informative non-invasive method and are of practical importance for early diagnosis of inflammatory periodontal disease and effectiveness of “Lysobact” drug in pregnant women with iron deficiency anemia.

Interleukin-1α and -1β assessment in the gingival crevicular fluid of periodontal patients with chronic hepatitis C.

The study assessed whether the increased production of interleukin-1α (IL-1α) and interleukin-1β (IL-1β), as a result of chronic hepatic inflammation, could be the expression of the negative impact on periodontal disease. The study included chronic periodontitis patients who were systemically healthy, chronic periodontitis patients suffering from chronic hepatitis C, as well as control patients, being systemically and periodontally healthy. After periodontal examination and the assessment of certain periodontal parameters, gingival crevicular fluid was collected from all participating patients. By using the enzyme-linked immunosorbent assay method, a quantitative assessment of IL-1α and IL-1β levels was possible. The immunologic results were correlated to the clinical periodontal data. The gingival fluid levels of cytokines were higher for periodontitis patients with chronic hepatitis C than for the systemically healthy periodontitis patients (1.8-fold higher for IL-1α and 2.1-fold higher for IL-1β). In addition, the gingival fluid cytokine levels were significantly higher for the periodontal patients (with/without chronic hepatitis C) than for the control group. Positive correlations were found between gingival fluid IL-1α and IL-1β levels and certain clinical periodontal parameters or the age of the viral hepatitis C diagnosis, in periodontitis patients with chronic hepatitis C. The chronic hepatic inflammation may have an important additional negative impact on the periodontal status, as both inflammatory reactions seem to be promoted by common pro-inflammatory cytokines.

IL-1β–Primed Mesenchymal Stromal Cells Improve Epidermal Substitute Engraftment and Wound Healing via Matrix Metalloproteinases and Transforming Growth Factor-β1

Since the 1980s, deep and extensive skin wounds and burns are treated with autologous split-thickness skin grafts, or cultured epidermal autografts, when donor sites are limited. However, the clinical use of cultured epidermal autografts often remains unsatisfactory because of poor engraftment rates, altered wound healing, and reduced skin functionality. In the past few decades, mesenchymal stromal cells (MSCs) have raised much attention because of their anti-inflammatory, protrophic, and pro-remodeling capacities. More specifically, gingival MSCs have been shown to possess enhanced wound healing properties compared with other tissue sources. Growing evidence also indicates that MSC priming could potentiate therapeutic effects in diverse invitro and invivo models of skin trauma. In this study, we found that IL-1β-primed gingival MSCs promoted cell migration, dermal-epidermal junction formation, and inflammation reduction invitro, as well as improved epidermal substitute engraftment invivo. IL-1β-primed gingival MSCs had different secretory profiles from naive gingival MSCs, characterized by an overexpression of transforming growth factor-β and matrix metalloproteinase (MMP) pathway agonists. Eventually, MMP-1, MMP-9, and transforming growth factor-β1 appeared to be critically involved in IL-1β-primed gingival MSC mechanisms of action.

ELISA Evaluation of RANKL Levels in Gingival Fluid in Patients with Periodontitis and Occlusal Trauma

The aim of this study was to evaluate the differences in RANKL levels in crevicular fluid (GCF) in patients with chronic periodontitis, with or without chronic occlusal trauma. The study group consisted of 40 patients from whom 72 samples of crevicular fluid were collected. RANKL levels were analyzed by ELISA. We noticed significantly higher differences in RANKL levels for the study group (occlusive trauma patients) than for systemic healthy patients (p = 8.008). Research has shown that secondary occlusal trauma associated with periodontal disease is characterized by significantly higher RANKL levels in patients with chronic occlusal trauma. This partially clarifies the molecular mechanisms that underlie more severe tissue destruction in patients with occlusal trauma.

IL-1β, TNF-α, and IL-6 levels in gingival fluid and serum of patients with ischemic stroke.

It has been documented that an increase of immune cells in periodontitis may stimulate the process of atherosclerosis and lead to ischemic stroke. The aim of this study was to investigate whether there is any significant difference in serum and gingival crevicular fluid (GCF) cytokine concentrations between patients with early ischemic stroke (IS) and a control group. Eighty-six patients were consecutively enrolled for this study, 50 with ischemic stroke and 36 as a control group with no previous stroke. Only dentulous patients were included. In each patient, GCF was collected from the deepest periodontal pocket. Blood serum samples were also taken from all subjects. The collected material was analyzed for cytokine concentrations using the cytometric bead array technique. Cerebral ischemia was found to be correlated with higher serum levels of IL-1β (13.01 ± 13.87 vs. 7.83 ± 2.11) and IL-6 (15.82 ± 16.64 vs. 6.64 ± 2.5) compared to the control group. The levels of IL-1, IL-6 and TNF-α in GCF were higher in patients with ischemic stroke but the difference was not significant. In conclusion, ischemic stroke was found to be associated with higher levels of IL-1β and IL-6 in serum. The cytokine concentrations in GCF did not appear to be directly correlated with those in serum in patients with ischemic stroke.(J Oral Sci 58, 509-513, 2016).

IL‐1 beta, IL‐12 and IL‐18 levels in gingival fluid and serum of patients with gingivitis and periodontitis

IL‐1 beta, IL‐12 and IL‐18 levels in gingival fluid and serum of patients with gingivitis and periodontitis

Interleukin‐1 β, interleukin‐12 and interleukin‐18 levels in gingival fluid and serum of patients with gingivitis and periodontitis

Cytokines are of major importance in periodontal disease progression. Interleukin-12 (IL-12) stimulates interferon-gamma production by T helper type 1 (Th1) cells while IL-18 induces Th1 responses when present with IL-12 but Th2 responses in the absence of IL-12. IL-1beta has been correlated with periodontal disease destruction. This study determined the local concentrations of these cytokines in sites of gingivitis and periodontitis. Gingival crevicular fluid was collected from two sites in each of 10 gingivitis patients and from two gingivitis sites and two periodontitis sites from each of 10 periodontitis patients. Serum samples were also collected. IL-1beta, biologically active IL-12 p70, the IL-12 p40 subunit and IL-18 concentrations were determined by enzyme-linked immunoabsorbent assay. IL-1beta and IL-18 concentrations were higher in the gingival crevicular fluid from periodontitis patients than in that from gingivitis patients; IL-18 concentrations were higher than those of IL-1beta. Very little IL-12, either p40 or p70, was detected in the gingival crevicular fluid samples. In the serum, very low levels of cytokines were found. The level of serum IL-12 p40, however, was higher than in the fluid from periodontitis sites of periodontitis patients. The local production of IL-1beta and IL-18 in the gingival crevicular fluid increased with increasing inflammation and IL-18 was the predominant cytokine at both gingivitis and periodontitis sites. Very little IL-12 was detected with levels decreasing with increasing inflammation. These results suggest that there is an association between severity of periodontal disease and levels of IL-1, IL-12 and IL-18.

Increased Interleukin‐18 in Patients With Juvenile Idiopathic Arthritis and Early Attachment Loss

Patients with juvenile idiopathic arthritis (JIA) have been shown to have incipient attachment loss (AL) more frequently than systemically healthy individuals. This study investigated neutrophil activity and proinflammatory cytokines in these patients and aged-matched controls. Elastase activity, measured with a low molecular weight substrate (S-2484), and interleukin-1beta (IL-1beta), measured with the enzyme-linked immunosorbent assay (ELISA), were analyzed in the gingival fluid of 38 patients with JIA and 29 controls. IL-1beta and interleukin-18 (IL-18) were measured with ELISA in the serum of the same groups. Subgingival plaque was analyzed by DNA probes to detect 12 bacteria. Significantly elevated serum levels of IL-1beta and IL-18 were found in the JIA group. No differences were found in the gingival fluid levels of elastase and IL-1beta between groups, or in the frequency of subjects positive to most of the bacteria analyzed, except F. nucleatum, C. rectus, P. micros, and S. intermedius, which were significantly more frequent in the control group. When the JIA group was subdivided according to the presence/absence of AL, IL-18 was significantly increased in the JIA subgroup with AL compared to those without it. There were no differences in the subgingival microbiota between the subgroups. The findings of increased serum IL-18 and IL-1beta in patients with JIA accompanied by a similar subgingival microbiota suggest that the increased frequency of incipient attachment loss observed in these patients might be due to their altered systemic inflammatory response, making them more susceptible to periodontal disease.

Accumulation of ciprofloxacin and minocycline by cultured human gingival fibroblasts.

Through a mechanism that is unclear, systemic fluoroquinolones and tetracyclines can attain higher levels in gingival fluid than in blood. We hypothesized that gingival fibroblasts take up and accumulate these agents, thereby enhancing their redistribution to the gingiva. Using fluorescence to monitor transport activity, we characterized the accumulation of fluoroquinolones and tetracyclines in cultured human gingival fibroblast monolayers. Both were transported in a concentrative, temperature-dependent, and saturable manner. Fibroblasts transported ciprofloxacin and minocycline with K(m) values of 200 and 108 micro g/mL, respectively, at maximum velocities of 4.62 and 14.2 ng/min/ micro g cell protein, respectively. For both agents, transport was most efficient at pH 7.2 and less efficient at pH 6.2 and 8.2. At steady state, the cellular/extracellular concentration ratio was > 8 for ciprofloxacin and > 60 for minocycline. Thus, gingival fibroblasts possess active transporters that could potentially contribute to the relatively high levels these agents attain in gingival fluid.

Genetic variations in cytokine expression: a risk factor for severity of adult periodontitis.

Periodontitis is a collection of chronic inflammatory diseases that are caused by specific bacteria. The bacteria activate inflammatory mechanisms in the periodontal tissues that destroy collagen and bone that support the teeth. Although bacteria are essential for the initiation of periodontitis, the quantity and types of bacteria have not been sufficient to explain the differences in disease severity. In recent years, it has become evident that for many common chronic diseases, there are modifying factors that do not cause the disease but rather amplify some disease mechanisms to make the clinical condition more severe. There are now data to suggest that a few factors which amplify the inflammatory process make people susceptible to an increased severity of periodontitis. Studies of untreated disease in Sri Lanka identified 3 patterns of disease progression. Studies in twins suggested that part of the clinical characteristics of periodontitis may be explained by genetic factors, but previous attempts to identify genetic markers for periodontitis have been unsuccessful Some genetic variations (polymorphisms) are commonly found in our population and represent a mechanism by which individuals may exhibit variations within the range of what is considered biologically normal. Since certain cytokines are key regulators of the inflammatory response and are important in periodontitis, we investigated the relationship between genetic variations associated with cytokine production and periodontitis severity. There are several polymorphisms in the cluster of genes that influence IL-1 biological activity. In recent clinical trials, two of these polymorphisms, when found together, have been associated with a significant increase in the risk for severe generalized periodontitis. Genetic association with periodontitis was evident only when smokers were excluded from the analysis, confirming the importance of smoking, and suggesting that both smoking and the IL- I genotype are independent factors in severe periodontitis. It is notable that 1 polymorphism associated with severe periodontitis in our study is also known to correlate with a 2- to 4-fold increase in IL-1 beta production. These findings are consistent with the current model of how genetic factors influence common chronic diseases. If we apply this model to periodontitis, it would involve the following: 1) a disease-initiating factor that would undoubtedly be specific bacteria such as Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans. and Bacteroides forsythus: and 2) modifiers of disease mechanisms that account for the clinical severity, including smoking, the IL-I genotype, certain systemic diseases, and psychosocial stress. The association of the IL-I genotype with severe periodontitis is consistent with several lines of periodontal research. Several studies have suggested there is a substantial genetic influence in periodontal disease. Although specific genetic markers have been identified in the uncommon juvenile forms of periodontitis, previous studies of specific genetic markers in adults with periodontitis have not been encouraging. Many investigators have, however, demonstrated a role for IL-1 in the initiation and progression of periodontitis. For example, IL-1 activates the degradation of the extracellular matrix and bone of the periodontal tissues, and elevated tissue or gingival fluid levels of IL-1 beta have been repeatedly associated with periodontitis. In addition, IL-1 is a strong enhancer of tissue levels of PGE2 and TNF-alpha. The association of severe periodontitis with smoking and the IL-1 genotype suggest a role for these factors in the pathogenesis of periodontitis. The finding that host modifying factors are associated with severe periodontitis suggest a biological mechanism by which some individuals, if challenged by bacterial accumulations, may have a more vigorous immunoinflammatory response, leading to more severe clinical disease. (ABSTRACT

Gingival fluid levels of acid and alkaline phosphatase.

Microtiter assays for acid and alkaline phosphatase levels in gingival fluid (GF) have been evaluated for their use as possible indicators of periodontal disease activity. Alkaline phosphatase concentration was shown to be positively associated with periodontal disease activity, while acid phosphatase showed no relation to disease activity. When a series of timed gingival fluid samples was taken from several sites in one subject's mouth, reproducible differences in volume, alkaline phosphatase concentration, and total acid phosphatase were found between the first and subsequent samples. Acid and alkaline phosphatase are presented as models of how substances can enter the periodontal pocket by different routes.

Concentration of doxycycline in human gingival fluid.

Doxycycline is a synthetic tetracycline compound whose main advantages over tetracycline hydrochloride are increased oral absorption, prolonged serum half-life and decreased gastrointestinal side-effects. The purpose of this study was to measure the concentration of doxycycline in gingival fluid and blood after oral administration. 4 volunteers were given doses of 100 mg doxycycline every 12 h on the first day of antibiotic administration followed by a maintenance dose of 100 mg per day for an additional 4 days. 3 of these volunteers were also given tetracycline hydrochloride every 6 h for 5 days either 1 month before or after doxycycline administration to compare gingival fluid levels of these 2 tetracycline compounds. Gingival fluid was sampled from 4 gingival sites in each volunteer at hourly intervals from hours 0 to 6, 9, 24, 27, 48 to 54, 57, 72, 75, 96 to 102 and 105. Blood was sampled by finger puncture at hours 0, 3, 6, 24, 48, 54, 72, 96 and 102. Antibiotic levels in gingival fluid and blood were measured using an agar diffusion assay method. The results demonstrated that doxycycline achieved much higher levels in the gingival fluid than in blood and yielded comparable gingival fluid levels to those achieved by tetracycline hydrochloride. Doxycycline levels in gingival fluid ranged between 1.2 micrograms/ml and 8.1 micrograms/ml in the first 24 h and generally achieved 3-10 micrograms/ml after 48 h. Blood levels after 48 h ranged between 2.1 micrograms/ml and 2.9 micrograms/ml. Tetracycline hydrochloride in gingival fluid after 48 h was generally in the range of 4 micrograms/ml-10 micrograms/ml with blood levels between 2.2 micrograms/ml and 3.4 micrograms/ml.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibiotic susceptibility of anaerobic bacteria from the human oral cavity.

Anaerobic, agar-dilution, minimal inhibitory concentrations (MICs) of 18 antibiotics are given for the numerically important bacterial groups from the human oral cavity. Strains are divided into susceptibility categories using the guidelines for interpretation of MICs suggested by the National Committee for Clinical Laboratory Standards. These guidelines are based on data on antibiotic concentrations attainable in serum following various dosage regimens. MICs are also compared with attainable gingival fluid levels where these are known. The highest percentages of strains were susceptible to tetracycline, with 89% of the 139 strains tested susceptible to serum levels and 97% conditionally susceptible to attainable gingival fluid levels. Ninety-eight percent of strains were conditionally susceptible to attainable gingival fluid levels of minocycline, but many strains, including Actinobacillus actinomycetemcomitans, were only moderately susceptible to attainable serum levels of this tetracycline analogue. Carbenicillin was effective against most groups of organisms, with the important exception of A. actinomycetemcomitans, at serum levels attainable with oral formulations of carbenicillin. Only 2% of the total strains tested were resistant to penicillin, while 33% of strains were categorized as moderately susceptible. Clindamycin was active against many strains of Gram-negative bacteria but was not active against A. actinomycetemcomitans, some Bacteroides, Eikenella corrodens, or the anaerobic vibrios. Metronidazole was active against A. actinomycetemcomitans, all five groups of oral Bacteroides tested, and against Capnocytophaga species. Chloramphenicol was active against A. actinomycetemcomitans, but not against most of the other groups of oral organisms. Nearly all groups contained strains non-susceptible to serum levels attainable with the usual doses of erythromycin, spiramycin, vancomycin, kanamycin, neomycin, streptomycin, doxycycline, oxytetracycline, or chlortetracycline; several strains were resistant to maximum attainable serum levels of each of these antibiotics except doxycycline.