leukotriene-b4 and Periodontitis

Neutrophils play a major role in the host response against invading periodontopathogenic microorganisms. Localized aggressive periodontitis (LAgP) is associated with various functional abnormalities of neutrophils. Based on the recent findings, LAgP neutrophils are not "hypofunctional" or "deficient." They are "hyperfunctional," and their amplified activity is responsible for the tissue destruction in periodontal disease. Several signal transduction abnormalities are associated with elevated neutrophil function in LAgP. There is a strong correlation between defective chemotaxis and decreased intracellular Ca2+ levels; total calcium-dependent protein Kinase C (PKC) activity of neutrophils is significantly lower than healthy subjects; and there is a marked increase in diacylglycerol (DAG) accompanied by a pronounced decrease in DAG kinase activity. In a separate set of experiments on the involvement of the inducible cyclooxygenase isoform (COX-2) and the role of novel lipid mediators in the pathogenesis of periodontal disease, crevicular fluid samples from LAgP patients were found to contain prostaglandin E2 (PGE2) and 5-LO-derived products, leukotriene B4 (LTB4), and the biosynthesis interaction product, lipoxin LXA4. Neutrophils from peripheral blood of LAgP patients, but not from healthy volunteers, also generated LXA4, suggesting that this immunomodulatory molecule may have a role in periodontal disease. Lipoxin generation and its relationship to PGE2 and LTB4 can be visualized as an important marker for the pathogenesis of periodontal disease. Thus, major advances in our understanding of the role of the neutrophil in host defense against periodontal organisms have been made through studies of LAgP. LAgP is used as an example of a severe periodontal disease that is related to abnormal neutrophil function. In this model, it appears that a hyperresponsiveness of the neutrophil, due to cell priming/predisposition, results in enhanced tissue damage.

Topics: Adjuvants, Immunologic; Biomarkers; Calcium; Chemotaxis, Leukocyte; Cyclooxygenase 2; Diacylglycerol Kinase; Diglycerides; Dinoprostone; Gingival Crevicular Fluid; Humans; Hydroxyeicosatetraenoic Acids; Isoenzymes; Leukotriene B4; Lipoxins; Membrane Proteins; Neutrophil Activation; Neutrophils; Periodontitis; Peroxidases; Prostaglandin-Endoperoxide Synthases; Protein Kinases; Signal Transduction; Stereoisomerism

The reported therapeutic benefits of nonsteroidal anti-inflammatory drugs (NSAIDs) in slowing periodontal disease progression appear intimately linked to the effective inhibition of local prostaglandin synthesis. This randomized, partially double-blind, controlled trial was conducted to evaluate the pharmacodynamic effects of the NSAID, ketoprofen (KTP), on gingival crevicular fluid (GCF) prostanoids. 42 subjects, ages 35-57 years, with moderate to advanced adult periodontitis were recruited and monitored for 22 days. On day 1, subjects were randomized for 1 of 5 treatments: i) 0.5% KTP gel; ii) 1.0% KTP gel; iii) 1.0% KTP alternate gel; iv) 2.0% KTP gel; v) 25 mg KTP capsule (positive control). Subjects applied 1 ml of gel topically to their gingiva or administered one capsule p.o., b.i.d. for 14.5 days. GCF samples were collected from posterior, interproximal sites on days 1 (pre-dosing; 1, 2, 3, 6 h), 8 (pre-dosing; 2 h), 15 (pre-dosing; 2 h) and 22 (post-treatment). GCF levels of prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) were determined using RIA, and expressed in ng/ml and % reduction from baseline (%Effect). Neither a significant difference among groups nor a dose response in % effect for either prostanoid was evident, both overall and among cohorts with elevated baseline mediator levels ([PGE2]>34 ng/ml; [LTB4]>300 ng/ml). When data were combined from all groups, significant (p<0.01) % reductions in GCF PGE2 were noted at 1 and 2 h post-dosing (29% and 24% respectively). In comparing topical versus systemic formulations, all topical formulations were as equipotent as systemic dosing in altering local prostaglandin levels despite lower KTP exposures with gel treatments. These data indicate that both topical and systemic KTP therapies pharmacodynamically reduce GCF PGE2 levels in adult periodontitis subjects, allowing for potential inhibition of disease progression.

Topics: Administration, Topical; Adult; Analysis of Variance; Anti-Inflammatory Agents, Non-Steroidal; Area Under Curve; Chi-Square Distribution; Dinoprostone; Dose-Response Relationship, Drug; Double-Blind Method; Female; Gingival Crevicular Fluid; Humans; Immunity, Cellular; Ketoprofen; Leukotriene B4; Male; Middle Aged; Periodontitis; Prostaglandins; Statistics, Nonparametric

The aim of this study was to measure tissue levels of immunoreactive prostaglandin E2 (iPGE2), immunoreactive leukotriene B4 (iLTB4), and pain after periodontal surgery and to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID), ibuprofen, on these levels. Two contralateral quadrants in each of nine patients were selected to undergo separate surgical procedures, one with ibuprofen (800 mg 1 hour presurgery and 400 mg postsurgery) and one with a placebo. Intra-operatively, a custom-made microdialysis probe, with a 3,000 dalton molecular weight cut-off, was inserted beneath the soft tissue flap and a dialysate collected every 20 minutes for 4 hours after surgery. Pain perception was measured at the same time intervals using two pain scales. Dialysate samples were assayed using two enzyme immunoassays. Mean tissue levels of iPGE2 in the placebo group increased from 74 nM at 40 minutes to a peak of 261 nM at 200 minutes. Mean tissue levels of iLTB4 in the placebo group fluctuated between 0.2 and 0.6 nM. Pain levels in this group increased continuously with time, peaking at 4 hours. Mean tissue levels of iPGE2 in the ibuprofen group were significantly suppressed, exhibiting more than a 95% reduction. This was accompanied by a significant reduction in pain. Ibuprofen had no detectable effect on tissue levels of iLTB4. These data indicate that iPGE2 and iLTB4 are present at relatively high concentrations in the periodontal tissues after surgery. Since these concentrations exceed the Kd values for binding to their respective receptors, PGE2 and LTB4 may be associated with the development of postsurgical pain and inflammation. These data also indicate that ibuprofen can successfully inhibit iPGE2 production in the periodontal tissues and in this way may help reduce postoperative pain and inflammation.

Topics: Adult; Analysis of Variance; Anti-Inflammatory Agents, Non-Steroidal; Binding Sites; Dinoprostone; Double-Blind Method; Extracellular Space; Female; Humans; Ibuprofen; Immunoenzyme Techniques; Leukotriene B4; Male; Microdialysis; Middle Aged; Pain Measurement; Pain, Postoperative; Periodontitis; Periodontium; Prostaglandin Antagonists; Statistics, Nonparametric

Here we determine the salivary levels of leukotriene B4 (LTB4 ) and its relation with salivary mucin and alveolar bone level.. LTB4 is a membrane-derived lipid mediator formed from arachidonic acid. It is among the most potent stimulants of polymorphonuclear leukocytes providing the first host defense against infections. Leukotrienes also induce bone resorption. Because LTB4 is present in the oral cavity the aim of the present study was to explore the role of LTB4 in patients with periodontal disease.. Eighty-one subjects were clinically examined and distributed into four groups, namely, clinically healthy, mild, moderate and severe periodontitis, according to periodontal status, classified into values of clinical attachment level and probing pocket depth. Unstimulated saliva was collected for 5 min. Salivary LTB4 was determined by an immune assay method, mucin was determined by a colorimetric method and radiographic assessment used to determine alveolar bone level.. Patients with mild periodontitis showed a decrease in salivary LTB4 levels while patients with severe periodontitis showed increased LTB4 levels. A significant positive correlation was observed between salivary LTB4 and clinical attachment level, salivary mucin concentration or alveolar bone level.. The close relation between salivary LTB4 and mucin levels suggested that LTB4 might be involved in the defense mechanism of the oral cavity. The correlation of LTB4 with the alveolar bone level indicates that they are one of the mediators responsible for bone resorption.

Topics: Adult; Alveolar Bone Loss; Alveolar Process; Colorimetry; Female; Humans; Leukotriene B4; Male; Middle Aged; Mucins; Neutrophils; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Periodontium; Radiography, Bitewing; Saliva; Salivary Proteins and Peptides

Resolution of inflammation is an active temporally orchestrated process demonstrated by the biosynthesis of novel proresolving mediators. Dysregulation of resolution pathways may underlie prevalent human inflammatory diseases such as cardiovascular diseases and periodontitis. Localized Aggressive Periodontitis (LAP) is an early onset, rapidly progressing form of inflammatory periodontal disease. Here, we report increased surface P-selectin on circulating LAP platelets, and elevated integrin (CD18) surface expression on neutrophils and monocytes compared to healthy, asymptomatic controls. Significantly more platelet-neutrophil and platelet-monocyte aggregates were identified in circulating whole blood of LAP patients compared with asymptomatic controls. LAP whole blood generates increased pro-inflammatory LTB4 with addition of divalent cation ionophore A23187 (5 µM) and significantly less, 15-HETE, 12-HETE, 14-HDHA, and lipoxin A(4). Macrophages from LAP subjects exhibit reduced phagocytosis. The pro-resolving lipid mediator, Resolvin E1 (0.1-100 nM), rescues the impaired phagocytic activity in LAP macrophages. These abnormalities suggest compromised resolution pathways, which may contribute to persistent inflammation resulting in establishment of a chronic inflammatory lesion and periodontal disease progression.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Blood Platelets; Calcimycin; Calcium Ionophores; Cells, Cultured; Chromatography, Liquid; Eicosapentaenoic Acid; Female; Humans; Hydroxyeicosatetraenoic Acids; Leukotriene B4; Lipoxins; Macrophages; Male; Middle Aged; Monocytes; Neutrophils; P-Selectin; Periodontitis; Phagocytosis; Tandem Mass Spectrometry

Resolvin E1 (RvE1) is a potent proresolving mediator of inflammation derived from omega-3 eicosapentaenoic acid that acts locally to stop leukocyte recruitment and promote resolution. RvE1 displays potent counter-regulatory and tissue-protective actions in vitro and in vivo. Periodontal disease is a local inflammatory disease initiated by bacteria characterized by neutrophil-mediated tissue injury followed by development of a chronic immune lesion. In this study, we report the treatment of established periodontitis using RvE1 as a monotherapy in rabbits compared with structurally related lipids PGE(2) and leukotriene B(4). PGE(2) and leukotriene B(4) each enhanced development of periodontitis and worsened the severity of disease. Promotion of resolution of inflammation as a therapeutic target with RvE1 resulted in complete restoration of the local lesion, and reduction in the systemic inflammatory markers C-reactive protein and IL-1beta. This report is the first to show that resolution of inflammation by a naturally occurring endogenous lipid mediator results in complete regeneration of pathologically lost tissues, including bone.

Topics: Alveolar Bone Loss; Animals; C-Reactive Protein; Chronic Disease; Dinoprostone; Disease Models, Animal; Eicosapentaenoic Acid; Homeostasis; Inflammation; Inflammation Mediators; Interleukin-1beta; Leukotriene B4; Neutrophil Infiltration; Neutrophils; Periodontitis; Rabbits

Leukotriene B(4) (LTB(4)) is a membrane-derived lipid mediator formed from arachidonic acid. LTB(4) is among the most potent stimulants of polymorphonuclear leukocytes (PMNs) and, thus, participates in tissue injury by recruiting PMNs in a pathophysiologic scenario of periodontal diseases. The aim of the present study was to assess the relationship between clinical parameters and concentrations of LTB(4) within gingival crevicular fluid (GCF) from inflamed gingiva and periodontitis sites before and after the treatment of periodontitis.. Sixty subjects were divided into three groups with 20 subjects in each group: healthy (group 1), gingivitis (group 2), and chronic periodontitis (group 3). Groups were based on periodontal disease index (PDI), clinical attachment loss (CAL), and radiographic evidence of bone loss. Group 4 consisted of the subjects in group 3 at 6 to 8 weeks after treatment, i.e., scaling and root planing (SRP). GCF samples collected from each patient were quantified for LTB(4) using an enzymatic immunometric assay. In addition, the correlation between in situ LTB(4) levels and clinical parameters was analyzed in each group.. The highest mean LTB(4) concentration in GCF was observed in group 3 (185.2 pg/microl), and the lowest was observed in group 1 (39.6 pg/microl). Its level in group 3 decreased to 79.35 pg/microl after treatment (group 4). Further, GCF LTB(4) levels in all groups showed a statistically significant positive correlation with PDI and CAL (P <0.005).. The substantial increase in GCF LTB(4) concentrations with the severity of periodontal disease and a concomitant decrease in its level following SRP in subjects with periodontitis suggest a possible role for LTB(4) in the progression of periodontal disease.

Topics: Adult; Biomarkers; Case-Control Studies; Dental Scaling; Disease Progression; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Leukotriene B4; Male; Periodontitis; Statistics, Nonparametric

The present study was planned to evaluate the individual and combined effects of selective cyclooxygenase-2 (COX-2) inhibitor, celecoxib, and omega-3 fatty acid on the gingival tissue levels of prostaglandin E2 (PGE2), prostaglandin F2alpha (PGF2alpha), leukotriene B4 (LTB4), and platelet activating factor (PAF) in endotoxin-induced periodontitis in rats.. Experimental periodontitis was induced by repeated injection of Escherichia coli endotoxin (LPS). Forty-four adult male Sprague-Dawley rats were divided into five study groups: saline control, LPS, celecoxib, omega-3 fatty acid, and combination celecoxib and omega-3 fatty acid. Celecoxib and omega-3 fatty acid were given either as a single agent or as a combination therapy during 14 days of the study period. At the end of the 2-week protocol, the rats were sacrificed, the gingival tissues were dissected and extracted, and the extracts were analyzed for PGE2, PGF2alpha, and LTB4 levels by enzyme immunoassay and for PAF levels by radioimmunoassay. The defleshed jaws were analyzed morphometrically for alveolar bone loss. Data were evaluated statistically by using parametric tests.. LPS injection resulted in significantly more bone loss than the saline controls (P<0.05) and significant elevations in the gingival tissue levels of all the analyzed mediators except PGF2alpha. Individual administration of celecoxib revealed significant reductions in PGE2 and PAF levels (P <0.05), while omega-3 fatty acid provided significant reduction in PGE2, PGF2alpha, and LTB4 levels compared to the LPS group (P <0.05). Combined administration of celecoxib and omega-3 fatty acid exhibited significantly lower values than those of the LPS group in all the analyzed membrane phospholipid mediators (P <0.05), which approximated the levels in the saline control group (P>0.05).. The results of the present study indicate that celecoxib and omega-3 fatty acid, when used individually, show a rather partial effect on the control of the analyzed mediators, but when combined they show a synergic effect and provide significant reductions in the gingival tissue levels of PGE2, PGF2alpha, LTB4, and PAF in LPS-induced experimental periodontitis. These findings may pioneer further clinical human studies investigating the possible place of celecoxib and omega-3 fatty acid in periodontal treatment.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Celecoxib; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Endotoxins; Fatty Acids, Omega-3; Leukotriene B4; Male; Periodontitis; Platelet Activating Factor; Pyrazoles; Rats; Rats, Sprague-Dawley; Sulfonamides

Host defense mechanisms are impaired in patients with congenital neutrophil (polymorphonuclear neutrophils (PMN)) defects. Impaired PMN chemotaxis is observed in localized aggressive periodontitis (LAP), a familial disorder characterized by destruction of the supporting structures of dentition. In the present studies, we sought evidence for molecular events underlying this aberrant human PMN phenotype. To this end, PMN transendothelial migration and superoxide anion generation were assessed with LAP patients and asymptomatic family members, as well as patients with other chronic mucosal inflammation. PMN from LAP patients showed decreased transmigration across vascular endothelial monolayers (18 +/- 12% of control, n = 4) and increased superoxide anion generation (358 +/- 37%, p = 0.003). Gene expression was analyzed using oligonucleotide microarrays and fluorescence-based kinetic PCR. cDNA microarray and kinetic-PCR analysis revealed diminished RNA expression of leukocyte-type diacylglycerol (DAG) kinase alpha in PMN from LAP patients (4.6 +/- 1.7 relative units, n = 6, p = 0.007) compared with asymptomatic individuals (51 +/- 27 relative units, n = 7). DAG kinase activity was monitored by DAG phosphorylation and individual DAG molecular species were quantified using liquid chromatography and tandem mass spectrometry-based lipidomics. DAG kinase activity was also significantly decreased (73 +/- 2%, p = 0.007) and correlated with increased accumulation of 1,2-diacyl-sn-3-glycerol substrates (p = 0.01). These results implicate defects in both PMN transendothelial migration and PMN DAG kinase alpha signaling as disordered functions in LAP. Moreover, they identify a potential molecular lesion in PMN signal transduction that may account for their aberrant responses and tissue destruction in this disease.

Topics: Adolescent; Adult; Cell Migration Inhibition; Chemotaxis, Leukocyte; Chronic Disease; Diacylglycerol Kinase; Diglycerides; Endothelium, Vascular; Female; Gene Expression Profiling; Humans; Intracellular Fluid; Leukotriene B4; Male; Neutrophils; Oligonucleotide Array Sequence Analysis; Periodontitis; Signal Transduction; Up-Regulation

Doxycycline has been widely used in periodontal treatment for its antimicrobial and anti-enzymatic effects. Recently, bisphosphonates have been shown to inhibit alveolar bone resorption. The aim of the present study was to evaluate the effects of doxycycline and the bisphosphonate alendronate on the gingival tissue levels of prostaglandin E2 (PGE2), prostaglandin F2alpha (PGF2alpha), leukotriene B4 (LTB4), and platelet-activating factor (PAF) in endotoxin-induced periodontal breakdown in rats.. Experimental periodontitis was induced by repeated injection of Escherichia coli endotoxin (LPS) and 44 adult male Sprague-Dawley rats were divided into five study groups as follows: LPS, doxycycline + LPS, alendronate + LPS, doxycycline + alendronate + LPS, and saline control. Doxycycline and alendronate were given either as a single agent or as a combination therapy during the 7-day study period. At the end of the 1-week protocol, the rats were sacrificed, the gingival tissues were dissected and extracted, and the extracts were analyzed for PGE2, PGF2alpha, LTB4, and PAF levels. The defleshed jaws were analyzed morphometrically for alveolar bone loss. Data were evaluated statistically by using parametric tests.. Alveolar bone loss measurements revealed significantly higher values in LPS, doxycycline + LPS, alendronate + LPS, and doxycycline + alendronate + LPS groups in comparison to the saline control group (P <0.05). Combined administration of doxycycline and alendronate exhibited the most prominent inhibition on gingival tissue levels of PGE2 and PGF2alpha (P<0.05). Doxycycline + alendronate + LPS group also significantly reduced LTB4 and PAF levels, although doxycycline provided the most reduction in the levels of these mediators (P <0.05).. Alendronate and/or doxycycline may provide significant inhibition of the major inflammatory mediators of periodontal tissue destruction, and combined administration of these agents may provide beneficial effects in periodontal treatment. However, this hypothesis must be further verified by clinical human trials before introducing its use in dental practice.

Topics: Alendronate; Alveolar Bone Loss; Animals; Anti-Bacterial Agents; Dinoprost; Dinoprostone; Doxycycline; Drug Therapy, Combination; Endotoxins; Gingiva; Leukotriene B4; Male; Periodontitis; Platelet Activating Factor; Rats; Rats, Sprague-Dawley

The aim of the present study was 1) to evaluate the possible effects of therapeutic usage of omega-3 fatty acid on the gingival tissue levels of prostaglandin E2 (PGE2), prostaglandin F2alpha (PGF2alpha), platelet activating factor (PAF), and leukotriene B4 (LTB4) in endotoxin-induced periodontitis in rats and 2) to investigate whether prophylactic usage provides any additional benefits to therapeutic doses of omega-3 fatty acid.. Experimental periodontitis was induced by repeated injection of Escherichia coli lipopolysaccharide (LPS). Thirty-six adult male Sprague-Dawley rats were divided into four study groups: 1) saline controls; 2) LPS; 3) therapeutic omega-3 fatty acid (TO3); and 4) prophylactic plus therapeutic omega-3 fatty acid (P + TO3) groups. In TO3 group, omega-3 fatty acid was given for 15 days following induction of experimental periodontitis. In P + TO3 group, omega-3 fatty acid was started 15 days before baseline, and then periodontitis was induced at baseline and omega-3 fatty acid was continued for 15 days after baseline. On day 15 after baseline, all rats were anesthetized and sacrificed. PGE2, PGF2alpha, and LTB4 levels in gingival tissue samples were analyzed by enzyme immunoassay and PAF levels were analyzed by radioimmonoassay. Data were evaluated statistically by using parametric tests.. LPS injection resulted in significant amount of bone loss (P<0.05). Neither therapeutic nor prophylactic plus therapeutic administration of omega-3 fatty acid with the doses and duration of therapy used in the present study was effective in preventing endotoxin-induced alveolar bone loss. TO3 group exhibited significant decreases in the gingival tissue levels of PGE2, PGF2alpha, LTB4, and PAF compared to the LPS group (P<0.05). PGE2 and PGF2alpha levels in TO3 group were similar to those of the saline group (P>0.05), while LTB4 and PAF levels were statistically higher than the saline group (P<0.05). Prophylactic plus therapeutic usage of omega-3 fatty acid provided similar levels of all these mediators to those of the saline controls (P>0.05).. Therapeutic omega-3 fatty acid significantly reduced the gingival tissue levels of PGE2, PGF2alpha, LTB4, and PAF in experimental periodontitis. Furthermore, prophylactic usage of omega-3 fatty acid provided additional beneficial effects to the therapeutic administration by decreasing the gingival tissue levels of these mediators to levels of healthy tissue. These findings should be verified by longitudinal clinical trials investigating clinical and biochemical periodontal parameters to better define the possible role of omega-3 fatty acids in periodontal treatment.

Topics: Alveolar Bone Loss; Analysis of Variance; Animals; Dinoprost; Dinoprostone; Endotoxins; Fatty Acids, Omega-3; Gingiva; Immunoenzyme Techniques; Inflammation Mediators; Leukotriene B4; Male; Periodontitis; Platelet Activating Factor; Radioimmunoassay; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric

Leukotriene B4 (LTB4), a product of the lipoxygenase pathway of arachidonic acid metabolism, exhibits numerous activities that can account for most of the features of host responses seen in periodontal diseases. The aim of the present study was to examine the role of LTB4 in the pathogenesis of specific periodontal diseases.. LTB4 levels were investigated in gingival crevicular fluid (GCF) and gingival tissue (GT) samples of 10 patients with chronic periodontitis (CP), 12 patients with generalized aggressive periodontitis (GAgP), 6 patients with localized aggressive periodontitis (LAgP), 6 patients with gingivitis (G), and 6 periodontally healthy subjects (H). Periodontal status was evaluated by measuring probing depth, gingival index, papillary bleeding index, and plaque index. LTB4 was extracted from the samples by solid-phase method using C18 cartridge and was purified by high performance liquid chromatographic method and then analyzed by radioimmunoassay.. All patient groups had significantly higher levels of GCF and GT LTB4 compared to the control group (P<0.005). The CP patients had the highest LTB4 levels compared to those in other patient groups (P<0.005). GAgP, LAgP, and G groups had similar amounts of GCF and GT LTB4 (P>0.005). When the data were expressed as concentration, the CP group was found to have higher concentration of LTB4, compared to that of control group (P<0.005). GAgP, LAgP, and G groups had similar LTB4 concentration compared to that of control group (P>0.005). No significant difference was found between GAgP, LAgP, and G groups (P>0.005). The CP group had higher LTB4 concentration compared to both GAgP and LAgP groups (P<0.005). Although the CP group had a higher GCF LTB4 concentration compared to G group, this difference did not reach significance (P>0.005). No significant correlation was found between GCF and GT LTB4 levels and clinical parameters.. The results of the present study indicate that LTB4 is likely to be an important mediator in regulating inflammatory responses in the human periodontal tissues. This lipid mediator may play an important role in the pathophysiology of periodontal disease.

Topics: Adult; Aggressive Periodontitis; Case-Control Studies; Chromatography, High Pressure Liquid; Chronic Disease; Female; Gingiva; Gingival Crevicular Fluid; Gingivitis; Humans; Leukotriene B4; Male; Middle Aged; Periodontitis; Radioimmunoassay; Statistics, Nonparametric

Arachidonic acid metabolites exert a variety of distinct biological effects on the initiation and resolution of inflammatory diseases and their measurements in tissue can be critical to evaluate their regulatory function during the course of inflammation and to supplement in vitro experiments. The aim of this study was the detection and quantitative analysis of four arachidonic acid metabolites in small-sized biopsies of human periodontal tissues. The biopsies were homogenized and injected directly into a single analytical column of a RP-HPLC system. Detection was performed by a photodiode array detector. Calibration was established by dilutions of authentic standards of prostaglandin E2 (PGE2), leukotriene B4 (LTB4), 12(R)-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), and 15(S)-hydroxy-5,8,11,13-eicosatetraenoic acid (15-HETE). A total of 38 specimens weighing between 19 and 191 mg (wet tissue) were analyzed (mean = 59.9 +/- 30.2 mg). The detection limits were 1 pg for LTB4 and 12-HETE, 0.5 pg for 15-HETE, and 10 ng for PGE2. The concentrations of PGE2 and LTB4 were significantly higher in inflamed than in healthy periodontal tissues (P = 0.0079; P = 0. 0114). 12-HETE was detected in one biopsy (30 pg/g); 15-HETE was not detected. This method of homogenization, extraction, and analysis of arachidonic acid metabolites by RP-HPLC appears to be well suited for studies of human oral biopsies. Only small tissue samples and minimal laboratory equipment were required for a sensitive analysis.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Adult; Arachidonic Acid; Biopsy; Calibration; Chromatography, High Pressure Liquid; Dinoprostone; Humans; Leukotriene B4; Periodontitis; Reference Standards

The arachidonic acid metabolites prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) are inflammatory mediators which are likely to be involved in the pathogenesis of periodontal disease. PGE2 mediates vasodilatation, increases vascular permeability, enhances pain perception by bradykinin and histamine, alters connective tissue metabolism and enhances osteoclastic bone resorption. LTB4 causes the accumulation of inflammatory cells in the inflamed sites, and degranulation of polymorphonuclear leukocytes.. To measure gingival crevicular fluid (GCF) levels of PGE2, LTB4 and periodontal health.. The periodontal condition of 24 subjects was evaluated on the basis of plaque index, gingival index, probing depth, and attachment level. GCF samples were collected from one or two site(s) of each sextant per subject and the volume was measured using Periotron 6000. Samples were then assayed for PGE2 and LTB4 using a competitive enzyme immunoassay. Mean PGE2 and LTB4 levels were determined for each subject and group means compared.. Significant differences in the levels of PGE2 and LTB4 were found between patients with periodontitis, and non-periodontitis individuals (P < 0.001). The PGE2/LTB4 levels were positively correlated with the clinical parameters (P < 0.01) and reduced markedly after phase 1 of the periodontal treatment (P < 0.01). The total amount and concentration (ng ml-1) of LTB4 was positively correlated with the gingival index (P < 0.01).. These results indicate that the levels of PGE2 correlated with the severity of the periodontal status, and the levels of LTB4 correlated with gingival inflammation. Thus, our data suggest that the total amounts of PGE2/LTB4 may be good indicators for periodontal inflammation.

Topics: Adult; Alveolar Bone Loss; Dental Plaque; Dental Plaque Index; Dental Scaling; Dinoprostone; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Immunoenzyme Techniques; Inflammation Mediators; Leukotriene B4; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Periodontium; Regression Analysis; Root Planing; Subgingival Curettage

Ketoprofen creams were evaluated for the treatment of periodontal disease in a placebo-controlled, double-blind study in the rhesus monkeys, Macaca mulatta. Two formulations containing ketoprofen (1%), with or without vitamin E, were evaluated against appropriate controls (8 monkeys per group). Two weeks prior to treatment, the animals received prophylaxis on only the left side of the mouth (spontaneous model). Selected teeth on the right side of the mouth were ligated (ligature model). The creams were administered to the gingiva once daily at a standard dose of 1.8 ml per monkey for 6 months. Clinical assessments were made 2 wk before initiation, at baseline and 1, 2, 3 and 6 months post-treatment. The clinical parameters included plaque formation, gingival redness, edema, bleeding on probing and Ramfjord Attachment Level measurements (RAL). Radiographs were taken at 2 wk before initiation, baseline and at 3 and 6 months post-treatment. Digital, subtraction radiography was used to measure vertical linear bone loss along the interproximal root surfaces of the left and right mandibular first molars. Gingival crevicular fluid (GCF) was collected for biochemical assays on PGE2, TxB2, LTB4, IL-1 beta and TNF alpha. There were no significant differences among groups with respect to gingival indices. Radiographic data demonstrated significant positive effects on bone activity in both groups treated with ketoprofen formulations with improvement over time in the ligature model (0.01 < or = p < or = 0.04). The placebo group exhibited bone loss of 1.96 +/- 0.48 and 1.40 +/- 0.56 mm per site at 3 and 6 months, respectively. The group treated with ketoprofen cream showed an apparent bone gain of 0.28 +/- 0.41 and 0.78 +/- 0.47 mm per site at 3 and 6 months, respectively. The group treated with ketoprofen cream containing vitamin E showed a mean bone loss of 0.41-0.48 mm per site at 3 months with improvement to an apparent bone gain of 0.31 +/- 0.44 mm per site at 6 months. The biochemical data demonstrated early and significant suppression of GCF-LTB4 by both ketoprofen formulations at 1 month, which preceded the significant suppression of GCF-PGE2 at 2 and 3 months in the ligature model (p < 0.003) and at 2 to 6 months in the spontaneous model (p < 0.02). We conclude that ketoprofen at 1% level in suitable topical vehicles can effectively inhibit GCF-LTB4 and GCF-PGE2 and positively alter alveolar bone activity in the ligature-induced model of periodontitis in the m

Topics: Administration, Topical; Alveolar Bone Loss; Animals; Anti-Inflammatory Agents, Non-Steroidal; Dental Plaque; Dinoprostone; Double-Blind Method; Edema; Emollients; Female; Follow-Up Studies; Gingival Crevicular Fluid; Gingival Hemorrhage; Gingivitis; Interleukin-1; Ketoprofen; Leukotriene B4; Macaca mulatta; Male; Molar; Periodontal Attachment Loss; Periodontitis; Placebos; Radiographic Image Enhancement; Random Allocation; Subtraction Technique; Thromboxane B2; Tumor Necrosis Factor-alpha; Vitamin E

Ligature-induced periodontitis was monitored for 6 months in eight Macaca mulatta monkeys to examine clinical status, radiographic bone level, and crevicular fluid (CF) levels of prostaglandin E2 (PGE2), thromboxane B2 (TxB2), interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha, and leukotriene B4 (LTB4). A split-mouth design was used, with eight ligated teeth and eight contralateral nonligated teeth which develop soft-chow-promoted (spontaneous) disease. Ligated sites experienced an average attachment loss of 0.94 mm per site and linear bone loss of 0.88 mm per site, with spontaneous-periodontitis sites experiencing approximately half the loss of ligated sites. The CF mediator levels showed increased levels of PGE2 and TxB2 at the ligated sites, as compared with the spontaneous sites, with no significant contralateral differences in the IL-1 beta or LTB4 responses. The concentrations of LTB4 in CF reached an early threefold peak over the baseline level at 1 month. By 2 months there was a statistically significant threefold elevation in CF-PGE2 in the ligated sites and a twofold elevation in the spontaneous sites as compared to the baseline level (P = 0.041 and 0.008, respectively). The monocyte product IL-1 beta increased sharply at 2 months and returned to the baseline level by 6 months at both ligated and nonligated sites. Tumor necrosis factor alpha in CF was below the limit of detection at all sites throughout the experiment (i.e., < 2 ng/ml). The selective elevation of both PGE2 and TxB2 in ligated sites, compared with levels in spontaneous sites, in the presence of similar levels of LTB4 and IL-1 beta provides further evidence that these molecules regulate the magnitude of the tissue-destructive response in progressive periodontitis.

Topics: Animals; Dinoprostone; Exudates and Transudates; Gingiva; Interleukin-1; Leukotriene B4; Macaca mulatta; Periodontitis; Thromboxane B2; Time Factors

THIS CASE REPORT DESCRIBES clinical and laboratory findings for a 60-year-old woman with recently diagnosed Crohn's disease and severe generalized periodontitis. Comparison of dental radiographs taken in 1975, in 1983, and at the time of our evaluation in 1986 revealed dramatic progression of alveolar bone loss over that period. Standard laboratory blood tests did not reveal any remarkable significant leukocyte abnormalities, but flow cytometric analysis of lymphocytes revealed a paucity of B cells stained with anti-light chain antibodies, and an increased proportion of T lymphocytes which were dully-stained with anti-CD5 monoclonal antibody. B cell function as determined by in vitro proliferative responsiveness to anti-IgM antibody was only about 50% of that observed with cells from two healthy normal subjects. Serum leukotriene B4 (LTB4) was elevated to 150% of normal values, in spite of the fact that the patient was taking a systemic anti-inflammatory drug which is known to reduce LTB4 levels. The microbial flora was highly mixed and included several putative periodontopathic bacteria. Therapy consisted of oral hygiene instruction, scaling and root planing, mucoperiosteal-flap curettage, extracoronal splinting, and selective extraction of three teeth. The periodontal status improved markedly with therapy. Possible relationships between the patient's immunological status, her Crohn's disease, and the severe periodontal breakdown are discussed.

Topics: B-Lymphocytes; Crohn Disease; Female; Humans; Leukotriene B4; Lymphocytes; Middle Aged; Neutrophils; Periodontitis