Periodontal tissue disposition of azithromycin in patients affected by chronic inflammatory periodontal diseases.

BACKGROUND: The recognition that periodontal diseases are associated with specific pathogens has led to interest in the use of antibacterial drugs for inhibition of these microorganisms. On these bases, the present study was aimed at evaluating the tissue distribution of the new macrolide antibiotic azithromycin in patients subjected to oral surgery for chronic inflammatory diseases of both marginal and periapical periodontium. METHODS: Thirty-two patients were treated with azithromycin 500 mg/day orally for 3 consecutive days, and drug concentrations in plasma, saliva, normal gingiva, and pathological periodontal tissues were evaluated. For this purpose, samples of blood, saliva, normal gingiva, granulation tissue, and radicular granuloma or cyst wall (from dentigerous cyst) were collected during oral surgery or 0.5, 2.5, 4.5, and 6.5 days after the end of pharmacological treatment; then, azithromycin levels were measured by a microbiological plate assay, using Micrococcus luteus NCTC 8440 as the indicator organism. RESULTS: The concentrations of azithromycin in plasma, saliva, normal gingiva, and pathological tissues reached the highest values 12 hours after the last dose (0.37+/-0.05 mg/l, 2.12+/-0.30 mg/l, 6.30+/-0.68 mg/kg, and 11.60+/-1.50 mg/kg, respectively) and then declined gradually. Consistent levels of the drug in normal gingiva and pathological tissues could be detected, however, up to 6.5 days, indicating that azithromycin was retained in target tissues for a long time after the end of treatment. Moreover, azithromycin levels in both normal gingiva and pathological tissues exceeded the minimum inhibitory concentrations of most pathogens involved in the pathophysiology of chronic inflammatory periodontal diseases. Notably, azithromycin levels in pathological tissues were significantly higher than those in normal gingiva 0.5, 2.5, and 4.5 days after the last dose. CONCLUSIONS: The present results indicate a marked penetration of azithromycin into both normal and pathological periodontal tissues, suggesting that azithromycin represents a promising option in both adjunctive and prophylactic treatments of chronic inflammatory periodontal diseases.     

Inhibition of human neutrophil phagocytosis and intracellular killing of yeast cells by fluoride

These processes depend upon the catabolism of glucose, which can be inhibited by fluoride (F-). We studied them with a radiolabelling technique using [3H]-uridine, which is incorporated into viable yeast cells, but hardly at all into neutrophils and dead or phagocytosed yeast cells. Lysis of neutrophils and treatment of the lysate with [3H]-uridine allowed estimation of intracellular killing of yeast cells. F- inhibited neutrophil phagocytosis and intracellular killing in a dose-related manner. Intracellular killing was significantly more sensitive to environmental F- than was phagocytosis. This effect on phagocytosis was not because of inhibition of opsonization of yeast, nor was it related to an effect on medium Ca2+.     

Evaluation of adipokines and inflammatory mediator expression levels in patients with periodontitis and peri-implantitis: a cross-sectional study

Clinical Oral Investigations - The aim of this study was to analyze the mRNA and protein expression of adiponectin, leptin, visfatin, tumor necrosis factor (TNF)-α, and interleukin (IL)-6...     

Temporal changes in inflammatory mediator concentrations in an adjuvant model of temporomandibular joint inflammation

AIMS: To determine temporal changes in the concentrations found in the temporomandibular joint (TMJ) and trigeminal ganglion of 3 specific classes of inflammatory mediators commonly linked with conditions of joint inflammation. The intent was to determine whether concentrations of the neuropeptide calcitonin gene-related peptide (CGRP), the neurotrophin nerve growth factor (NGF), and the proinflammatory cytokines interleukin-1beta (IL1beta) and tumor necrosis factor-alpha (TNF-alpha) are altered in the trigeminal ganglion and TMJ tissues during various stages of adjuvant-induced inflammation of the rat TMJ. METHODS: Adult male rats received bilateral TMJ injection of complete Freund's adjuvant (CFA), while control rats did not receive CFA treatment. The trigeminal ganglion and TMJ tissues were collected at 2 days, and 2, 4, and 6 weeks postinjection and analyzed using either radioimmunoassay or enzyme-linked immunosorbent assay. RESULTS: In the trigeminal ganglion, both CGRP and NGF concentrations were significantly elevated in comparison to controls from 2 days to 4 weeks; however, the patterns of increase differed. Concentrations of each inflammatory mediator were significantly elevated in the TMJ tissues of CFA-injected animals at 2 days and continued to be significantly elevated throughout the 6-week period. CGRP content remained at peak levels from 2 days through 6 weeks, while peak content for NGF, IL-1beta, and TNF-alpha was found at 2 days through 2 weeks. CONCLUSION: The results suggest that the development of CFA-induced inflammation of the TMJ was accompanied by a variable increase in the concentration of different classes of inflammatory mediators in both the trigeminal ganglion and TMJ tissues, which implies that each class of inflammatory mediator may play a significant role during different stages in the onset and exacerbation of the inflammatory process.     

Inhibition of VEGF gene expression in osteoblast cells by different NSAIDs

ObjectiveTo determine the effect of different nonsteroidal anti-inflammatory drugs (NSAIDs) on vascular endothelial growth factor (VEGF) gene expression in two osteoblast cell populations.DesignOsteoblasts obtained by primary culture (HOp) and human osteosarcoma cell line MG63 (MG-63), which were treated with 10 μM doses of acetaminophen, indomethacin, ketoprofen, diclofenac, ibuprofen, ketorolac, naproxen or piroxicam. At 24 h of treatment, their gene expression of VEGF was evaluated by real-time polymerase chain reaction (RT-PCR) and compared with the expression in untreated cells (control group).ResultsThe treatment with the different NSAIDs significantly reduced VEGF expression regardless of the cell line and NSAID studied.ConclusionThe results of this study suggest that these drugs may have undesirable effects on the osteoblast and its bone-forming capacity, given the effect of this growth factor on these cells. Further studies are warranted to determine their repercussions on bone tissue and to elucidate the cell signaling mechanism/s involved.     

Periodontitis-associated up-regulation of systemic inflammatory mediator level may increase the risk of coronary heart disease

Nakajima T, Honda T, Domon H, Okui T, Kajita K, Ito H, Takahashi N, Maekawa T, Tabeta K, Yamazaki K. Periodontitis‐associated up‐regulation of systemic inflammatory mediator level may increase the risk of coronary heart disease. J Periodont Res 2009; doi: 10.1111/j.1600‐0765.2009.01209.x. © 2009 John Wiley & Sons A/S Background and Objective: Although an elevation in the concentration of high‐sensitivity C‐reactive protein (hs‐CRP) as a result of periodontal infection may account for an increased risk of developing coronary heart disease (CHD), the effect of periodontal infection on the level of hs‐CRP in an otherwise healthy Japanese population has not yet been reported. The aim of the present study was to confirm, on a larger scale, our previous pilot study findings that both chronic periodontitis and subsequent periodontal treatment alter the serum levels of C‐reactive protein (CRP), interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α). Material and Methods: The concentrations of serum hs‐CRP, IL‐6 and TNF‐α were measured in 78 periodontitis patients at baseline and at re‐assessment, and in 40 periodontally healthy subjects at the time of examination. Results: The concentrations of hs‐CRP and IL‐6 in the sera of periodontitis patients were significantly higher than those in control subjects. By contrast, the concentration of TNF‐α was significantly lower in periodontitis patients than in control subjects. Whereas periodontal treatment decreased the levels of serum hs‐CRP and IL‐6, no such effect was observed for TNF‐α. When the patients were subdivided into four groups according to their initial concentration of hs‐CRP, only the CRP and IL‐6 concentrations of the highest quartile group showed a significant reduction following periodontal treatment. No significant difference in the initial clinical parameters was observed in any quartile. Conclusion: Although periodontal infection does affect the concentration of hs‐CRP and IL‐6 in serum, a subgroup of patients exist who are highly susceptible to an increased risk of CHD associated with periodontitis, suggesting that there may be subjects who have an elevated risk of CHD independent of susceptibility to periodontal tissue destruction per se.     

Lymphokine regulation of neutrophil phagocytic activity in patients with inflammatory diseases of the periodontium

The authors have demonstrated the possibility of a purposeful correction of the immune system disorders in patients with periodontitis, one of such disorders being a lowered phagocytic activity of neutrophils. Lymphokines, immune system mediators, were used for in vitro regulation of neutrophilic phagocytosis. The effects of lymphokines obtained from the peripheral blood lymphocyte culture supernatants of normal subjects and periodontitis patients, and of the purified lymphokine M- and L-fractions normalizing the reduced neutrophilic phagocytosis in periodontitis were under study.     

Alterations of neutrophil l-selectin and CD18 expression by tobacco smoke: implications for periodontal diseases

Alterations in neutrophil functions by both chronic low levels of tobacco and by acute short-term higher levels of tobacco smoke, as encountered during the act of smoking, may play a role in the pathogenesis of periodontal diseases in smokers. Among the early migration events of neutrophil function is the alteration in surface expression of L-selectin and the CD11/18 integrins. In the present study we examined the effect of in vitro smoke exposure and nicotine alone on the expression of these 2 adhesion molecules in neutrophils from smokers and non-smokers. We also determined the physiological relevance of this in vitro system by assessing the levels of nicotine exposure in this in vitro system and comparing these levels to acute and chronic levels of nicotine in saliva and gingival crevicular fluid. Peripheral neutrophils were isolated from the blood of smokers (< 1 pack/d) and non-smokers and incubated in vitro with either cigarette smoke (0-5 min), 10^?7 M F-met-leu-phe, or nicotine alone at 1.62 mg/ml to 162 ng/ml (10^?2 M-10^?6 M). The neutrophils were then incubated with fluoresceine conjugated anti-Leu8 (L-selectin), anti-CD 18 (CD 18 integrin), or γ-4 (non-specific control), fixed and analyzed by flow cytometry. With cigarette smoke exposure, there was an approximate 75% shedding of L-selectin in both smokers and non-smokers with no marked difference between groups at 1-5 min of smoke exposure. Cigarette smoke exposure resulted in a 15-20% increase in CD18 expression in both smokers and non-smokers. At all time points, there was slightly greater but statistically insignificant expression of CD 18 integrin in non-smokers when compared to smokers. These patterns of CD 18 increases and L-selectin shedding were similar in magnitude to incubations with 10^?7 M F-met-leu-phe. Acute smoke exposure resulted in elevation of nicotine in the smoke box to 529 ng/ml at 5 min, in saliva from 109.2 ng/ml before smoking to 1821.4 ng/ml after smoking, and in gingival crevicular fluid to 5961 ng/ml after smoking. No significant alterations in L-selectin or CD 18 expression were noted with in vitro nicotine from 1.62 mg/ml to 162 ng/ml.     

Alterations of neutrophil L-selection and CD18 expression by tobacco smoke: implications for periodontal diseases

Alterations in neutrophil functions by both chronic low levels of tobacco and by acute short-term higher levels of tobacco smoke, as encountered during the act of smoking, may play a role in the pathogenesis of periodontal diseases in smokers. Among the early migration events of neutrophil function is the alteration in surface expression of L-selectin and the CD11/18 integrins. In the present study we examined the effect of in vitro smoke exposure and nicotine alone on the expression of these 2 adhesion molecules in neutrophils from smokers and non-smokers. We also determined the physiological relevance of this in vitro system by assessing the levels of nicotine exposure in this in vitro system and comparing these levels to acute and chronic levels of nicotine in saliva and gingival crevicular fluid. Peripheral neutrophils were isolated from the blood of smokers (> 1 pack/d) and non-smokers and incubated in vitro with either cigarette smoke (0–5 min), 10^?7 M F-met-leu-phe, or nicotine alone at 1.62mg/ml to 162ng/ml (10^?2 M-10^?6 M). The neutrophils were then incubated with fluoresceine conjugated anti-Leu8 (L-selectin), anti-CD18 (CD18 integrin), or γ-4 (non-specific control), fixed and analyzed by flow cytometry. With cigarette smoke exposure, there was an approximate 75% shedding of L-selectin in both smokers and non-smokers with no marked difference between groups at 1–5 min of smoke exposure. Cigarette smoke exposure resulted in a 15–20% increase in CD 18 expression in both smokers and non-smokers. At all time points, there was slightly greater but statistically insignificant expression of CD18 integrin in non-smokers when compared to smokers. These patterns of CD18 increases and L-selectin shedding were similar in magnitude to incubations with 10^?7 M F-met-leu-phe. Acute smoke exposure resulted in elevation of nicotine in the smoke box to 529 ng/ml at 5 min, in saliva from 109.2 ng/ml before smoking to 1821.4 ng/ml after smoking, and in gingival crevicular fluid to 5961 ng/ml after smoking. No significant alterations in L-selectin or CD 18 expression were noted with in vitro nicotine from 1.62 mg/ml to 162 ng/ml.     

Doxycycline reduces lipopolysaccharide-induced inflammatory mediator secretion in macrophage and ex vivo human whole blood models

Background: Tetracyclines have been extensively used as adjuncts in the treatment of some forms of periodontitis. The aim of this study was to evaluate the capacity of doxycycline to influence the secretion of inflammatory mediators in macrophage and ex vivo human whole blood models stimulated with periodontopathogen lipopolysaccharides (LPS). Methods: Monocyte-derived macrophages were treated with various concentrations of doxycycline prior to being stimulated with Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) LPS. The capacity of doxycycline to mediate the inflammatory response was also tested in an ex vivo whole blood model (whole blood isolated from periodontitis patients and healthy subjects) stimulated with Porphyromonas gingivalis LPS. The secretion of interleukin (IL)-1beta, -6, and -8 and tumor necrosis factor-alpha (TNF-alpha) in both models was assessed by enzyme-linked immunosorbent assays (ELISA). Changes in phosphorylation state of kinases induced by A. actinomycetemcomitans LPS and doxycycline in the macrophage model were characterized by a multiplex ELISA analysis. Results: The secretion of IL-1beta and -8 and TNF-alpha by macrophages decreased significantly (P <0.05) when they were pretreated with 2 muM doxycycline, whereas a concentration of 10 muM was required to significantly reduce IL-6 secretion. Pretreatment of macrophages with 10 muM doxycycline prior to A. actinomycetemcomitans LPS stimulation resulted in a marked decrease in the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 (-76%). In the whole blood model, doxycycline, more particularly at 10 muM, was also a potent inhibitor of the proinflammatory cytokine response. Conclusion: These two models provided clear evidence that some of the clinically proven benefits of doxycycline may be related to its ability to regulate inflammatory mediator release by host cells.     

The expression of macrophage and neutrophil elastases in rat periradicular lesions

Macrophage elastase and neutrophil elastase are involved in tissue destruction in periradicular lesions. The purpose of this study was to examine these elastases immunohistochemically during development of periradicular lesions induced in rat mandibular first molar by pulpal exposure for 7, 14, 21, 28, and 42 days. Histologically, periapical inflammation developed from 7 to 21 days and then subsided after 28 days. The area of these lesions gradually increased from 7 to 28 days and subsequently decreased at 42 days. Macrophage elastase was first detected at 7 days and predominantly shown from 14 to 28 days, whereas neutrophil elastase gradually increased from 14 to 28 days. Macrophage elastase was significantly greater than neutrophil elastase from 7 to 21 days. These results suggest that macrophage elastase was enhanced from an early stage during the development of these lesions and that neutrophil elastase was related to the expansion of periapical tissue destruction including bone resorption.     

Suppression of human neutrophil functions by tetracyclines

Tetracycline inhibition of neutrophil-associated collagenolysis has been the focus of a number of investigations. Evidence has suggested that this inhibition results from the ability of this family of antimicrobial drugs to bind divalent cations such as Ca2+ and Zn2+, two cations that are required for full expression of activity of metalloproteinases such as collagenase and gelatinase. Data presented in this study demonstrate that tetracyclines can also inhibit neutrophil-mediated RBC lysis, superoxide anion synthesis, degranulation and migration. To some extent, tetracycline inhibition of neutrophil functions is mimicked by the Ca2+ binding agents, EDTA and TMB-8. However, Ca2+ enrichment restored full function to EDTA- and TMB-8-treated cells but not to tetracycline-treated neutrophils. This suggests that Ca2+ binding plays a role but is not the critical effect leading to tetracycline suppression of neutrophil functions. It has been suggested that tetracyclines can suppress leukocyte-associated tissue damage. Host tissues are protected from neutrophil-mediated damage by two mechanisms: 1. Neutrophil granule-associated enzymes are secreted in an inactive state; and, 2. tissues are protected from these enzymes by a potent inhibitor shield. Neutrophils can bypass these protective elements by activating enzymes and by destroying the shield through the synthesis of oxygen radicals. Therefore, tetracyclines may suppress neutrophil-mediated tissue damage by inhibiting their migration and degranulation and, potentially more importantly, by suppressing synthesis of oxygen radicals.(ABSTRACT TRUNCATED AT 250 WORDS)