Gingival crevicular fluid osteocalcin in adult periodontitis

This study aimed to detect the levels of osteocalcin in gingival crevicular fluid (GCF) from healthy (< or =3 mm sulcus depth and non-bleeding) and diseased sites (> or =6 mm probing depth and bleeding) in subjects with adult periodontitis, in order to further investigate its potential as a possible marker of the disease process. Periodontal probing depths, attachment levels and gingival indices were recorded from one healthy and one diseased site in each of 20 subjects with adult periodontitis. Both GCF accumulated in the periodontal pocket or sulci and GCF flowing into the periodontal pocket or sulci over a three-minute interval were sampled. The amounts of osteocalcin in each GCF sample was determined using immunoassays. A mean of 2.34 ng/site (2.7 microg/ml) osteocalcin was found at diseased sites and a mean of 2.47 ng/site (5.47 microg/ml) was found at healthy sites for the accumulated GCF collection method. A mean of 0.17 ng/ site (2.17 microg/ml) osteocalcin was found at diseased sites and a mean of 0.14 ng/ site (1.85 microg/ml) at healthy sites for the flow method of GCF collection. There were no statistically significant differences between osteocalcin levels in diseased and healthy sites in subjects with adult periodontitis.     

Vascular endothelial growth factor in human periodontal disease

Vascular endothelial growth factor (VEGF) is a multifunctional angiogenic cytokine of importance in inflammation and wound healing but its presence in chronic inflammatory periodontal disease has never been reported. The aims of this study were to investigate the presence of VEGF in human periodontal tissue and gingival crevicular fluid (GCF) in periodontal health and disease. VEGF in tissue was localized by immunohistochemistry. GCF and unstimulated saliva were collected from patients and clinically healthy subjects and VEGF was assessed by using an ELISA. VEGF was detected within vascular endothelial cells, neutrophils, plasma cells and junctional, pocket and gingival epithelium. In periodontitis patients, the volume of GCF and total amount of VEGF collected from diseased sites were both greater than from clinically healthy sites (Wilcoxon test p <0.01). However, the concentration of VEGF per unit volume of GCF was higher at healthy sites compared with diseased sites (Wilcoxon test p<0.05). Higher concentrations of VEGF were detected in healthy sites in patients compared with similar sites in clinically healthy subjects (Mann-Whitney U-test p <0.05). A logistic regression approach indicated that there was variation in VEGF between subjects (p<0.01), and that age (p<0.05), plaque (p < 0.05) and pocket depth (p < 0.07) were explanatory variables. VEGF was also detected in all saliva samples and was significantly higher in patients than in healthy controls (p<0.05). This study suggests that VEGF could be relevant to angiogenic processes in healthy as well as diseased periodontal tissue and that the periodontal status influences the salivary level of VEGF.     

Microbial factors and gingival crevicular fluid aspartate aminotransferase levels

Abstract. The aim of this cross-sectional study was to investigate the clinical application of chairside tests for gingival crevicular fluid (GCF) aspartale amino-transferase (AST) levels and plaque BANA hydrolysis activity with the presence of the periodontal pathogens Porphyromonas gingivalis and Actinohacillus action-mycetemcomitans. The study comprised 100 periodontitis sites (pocket depths≥4 mm. GI = 3) from 10 patients with chronic adult periodontitis and 100 control sites (pocket depths <4 mm. GI<3) from 10 periodontally healthy patients comprising 55 healthy sites (pocket depths <4 mm. GI=0) and 45 gingivitis sites (pocket depths <4 mm, GI=1 or 2). The values for both BANA hydrolysis and AST levels were significantly higher in samples from periodontitis compared with gingivitis and healthy sites (p<0.001), A. actinomycetemcomitans was identified in 45% and P. gingivalis in 17% of periodontitis sites but neither pathogen was recovered from control sites and there was no significant correlation with (he clinical parameters measured. There was no significant relationship between the presence of P. gingivalis and/or A. actinmycetemcomitans with BANA hydrolysis or AST levels. A significant correlation (p=0.0017) was observed between BANA hydrolysis and pocket depth and between AST hydrolysis and the GI (p=0.01). This study failed to demonstrate a positive association between chairside analysis of GCF metabolites for AST levels and/or BANA hydrolysis with P. gingivalis and A. actinomycetemcomitans. However, the GCF metabolites had a significant correlation with periodontally diseased sites in patients with chronic adult periodontitis and may help confirm clinical observations.     

Matrix metalloproteinase‐8 concentration in shallow crevices associated with the extent of periodontal disease

Objective: The aim of this study was to analyse the association between matrix metalloproteinase‐8 (MMP‐8) concentration in shallow, mostly non‐bleeding gingival crevices, and the extent of periodontal disease. Material and Methods: Plaque, bleeding on probing (BOP), probing pocket depth (PPD) and attachment level (AL) were assessed clinically in 48 patients with chronic periodontitis. MMP‐8 concentrations in gingival crevicular fluid (GCF) from four shallow (PPD3 mm), and four diseased sites and in serum, were measured by enzyme‐linked immunosorbent assay. Results: The mean concentration of MMP‐8 in GCF from shallow crevices was 11.8 ± 12.8 ng/ml and from diseased sites was 150.1 ± 91.8 ng/ml. In subjects with moderate to high plaque scores, a statistically significant association was found between MMP‐8 concentration from shallow crevices and the extent of AL4 mm (p=0.028) and AL6 mm (p<0.001). Conclusion: The above association between MMP‐8 concentration in shallow crevices and attachment loss provides a new aspect to future studies of MMP‐8 as a prognostic marker for periodontal disease.     

Relationship between levels of aspartate aminotransferase in gingival crevicular fluid and conventional measures of periodontal status assessed using PocketWatch: a cross-sectional study

The aim of this cross-sectional study was to determine, using PocketWatch, the relationship between the level of aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) and conventional measures of periodontal status, such as probing depth, attachment level, bleeding on probing and gingival index, in patients with untreated chronic periodontitis. A total of 15 patients with chronic periodontitis were enrolled. Their periodontal status and AST levels in their GCF were measured (n = 93) and statistically analyzed. There was a statistically significant difference in AST levels between diseased periodontal sites and healthy sites (p < 0.0001). The coefficients of correlation between AST levels and probing depth, attachment level and gingival index at all sites were 0.436, 0.266 and 0.468 (Spearman rank correlation). The correlation coefficients were too small to show a definite relationship between AST levels and individual measures of clinical periodontal status. However, AST levels may help to confirm clinical observations in patients with chronic periodontitis before therapy, since AST levels differentiate active and inactive periodontal diseased sites.     

Osteopontin in gingival crevicular fluid

Osteopontin (OPN) is a major glycosylated phosphoprotein in bone matrix and is produced by several cells including osteoblasts, osteoclasts and macrophages. OPN levels increase in active sites of bone metabolism. Recently, several bone-related proteins were identified in gingival crevicular fluid (GCF) to seek markers of alveolar bone resorption in periodontal disease. In this study, we investigated the existence of OPN in GCF and the correlation between OPN level in GCF and probing depth (PD) of sampling sites in 98 periodontitis patients and 35 healthy subjects. An immunoblotting analysis using 10% polyacrylamide gel showed that two forms of OPN with molecular masses of 54 and 66 kDa and several degraded fragments were detected in most GCF samples from diseased sites (PD > 4 mm). In GCF samples from healthy sites (PD < or = 3 mm), only one form (54 kDa) was observed, but any degraded fragments were not detected. When OPN amounts in GCF samples were determined by ELISA, a weak. but significant correlation was observed between OPN amount in GCF and PD (r=0.32, p=0.0013). These results demonstrate that OPN exists in GCF and that OPN level in GCF increases with the progression of periodontal disease.     

Five parameters of gingival crevicular fluid from eight surfaces in periodontal health and disease

Volume and amounts of myeloperoxidase (MPO), lactoferrin (LF), aryl sulfatase (AS) and lactate dehydrogenase (LDH) were measured in gingival crevicular fluid (GCF) collected from the mesial and distal proximal surfaces of the premolars and first and second molars of 3 subject groups. Group assignment was based on subject mean gingival index (GI) and probing depth (PD) of sampled sites as follows: healthy, GI less than or equal to 0.5, PD less than or equal to 3.0; disease 1, GI greater than or equal to 1.0, PD greater than or equal to 3.0 mm; disease 2, PD greater than or equal to 4.0 mm. Attachment loss (ATL) of most sites in the 3 groups was: healthy, 0-1 mm; disease 1, 1-2 mm; and disease 2, 4-9 mm. GCF volume differed among surfaces and teeth in each of the 3 groups. The greater amount of GCF collected from posterior locations was not related to the GI and PD. Differences with sampling location in amounts of GCF constituents were restricted to MPO and LF. Most of these differences (greater amounts at posterior sites) were associated with more severe disease. Variability in amount and composition of GCF collected from different sites, therefore, should be considered in experiments which include quantitation of GCF parameters. The ratio of MPO in disease group 2 to disease group 1 was greater than similar ratios for GCF volume and LF, AS and LDH. The quantity of MPO was the only measure which differed between the 2 disease groups at all surfaces. MPO thus appears to have the greatest potential, among the measured parameters, to serve as a marker for advanced periodontal disease.     

Radioimmunoassay quantification of adrenomedullin in human gingival crevicular fluid

OBJECTIVE: To investigate whether adrenomedullin (ADM), a multifunctional peptide with key roles in host antimicrobial defence and inflammation, was present and quantifiable in human gingival crevicular fluid (GCF) and to study its relationship with periodontal health and disease. DESIGN: GCF samples (30s) were collected using perio-paper strips from one diseased site in 21 subjects with periodontal disease and one healthy site from 19 control subjects with no evidence of periodontal disease. Samples were analysed by radioimmunoassay using a specific anti-human ADM antibody. RESULTS: Measurable adrenomedullin-like immunoreactivity (ADM-LI) was present in all the GCF samples collected. ADM-LI was significantly higher in periodontitis sites (mean 493.6 pg) than in control healthy sites (mean 248.5 pg), p = 0.0016. CONCLUSION: It is concluded that ADM is present in GCF at levels at which it could have an antibacterial role in the gingival crevice and modulate the pathophysiology of periodontal inflammation.     

Analysis of aspartate aminotransferase in gingival crevicular fluid assessed by using PocketWatch: a longitudinal study with initial therapy

AIM: The aim of this longitudinal study was to analyze the level of aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) by using PocketWatch before and after initial therapy in patients with chronic adult periodontitis and to determine the relationship between AST and conventional measures of periodontal status, such as probing depth, clinical attachment level, bleeding on probing, and gingival index. METHOD: A total of 11 patients with chronic adult periodontitis were enrolled. Their periodontal status and AST levels in GCF were measured at baseline and post-initial therapy (the number of pockets=67), and statistically analyzed. RESULTS: There was a statistically significant difference in AST levels between diseased periodontal sites (1.2+/-0.7) and healthy sites (0.3+/-0.6, p<0.05), and between baseline and post-initial therapy (p<0.05). Improvements in clinical status were noted following periodontal therapy and there was a corresponding decrease in AST levels. CONCLUSIONS: In conclusion, it is suggested that AST levels may be a useful adjunct in the clinical assessment of periodontal disease sites, since AST level decreases when periodontal status improves.     

Effect of periodontal treatment on IL-1beta, IL-1ra, and IL-10 levels in gingival crevicular fluid in patients with aggressive periodontitis

Aim: The aim of this study was to examine the effect of phase I periodontal treatment on the levels of interleukin (IL)‐1β, IL‐1ra, and IL‐10 in gingival crevicular fluid (GCF) in patients with generalized aggressive periodontitis (G‐AgP). Material and Methods: Data were obtained from 15 patients with aggressive periodontitis and 15 healthy controls. GCF was collected from at least four pre‐selected sites (one shallow, at least two moderate, or at least one deep pockets) in patients with G‐AgP. In the healthy group, GCF samples were collected from one site. The cytokine levels were determined by an enzyme‐linked immunosorbent assay. Probing depth, clinical attachment level (CAL), gingival and plaque indices, and bleeding on probing were measured. The GCF sampling and clinical measurements were recorded at baseline and 6 weeks later after periodontal treatment. Results: IL‐1β levels were significantly higher at the moderate and deep pocket sites compared with the shallow sites (p<0.05). After periodontal therapy, IL‐1β levels were significantly reduced in the moderate and deep pocket sites (p<0.05). IL‐1ra levels at baseline of the moderate and deep pocket sites were significantly lower than the control sites (p<0.05). IL‐10 levels were similar in all pockets and did not change after periodontal therapy. Conclusions: The periodontal treatment improves the clinical parameters in G‐AgP, and this improvement is evident in deep pocket sites for pocket depth and CAL values. These results confirm that IL‐1β is effective for evaluating the periodontal inflammation and can thus be used as a laboratory tool for assessing the activity of periodontal disease.     

Repeated measurement of crevicular fluid parameters at different sites

Few systematic studies have been made of amounts or of the composition of gingival crevicular fluid (GCF) from different sites or of the stability of GCF parameters over time. These data are needed to better understand the relation of GCF composition to periodontal health status. This report gives the volume and the amounts of lactate dehydrogenase (LDH), aryl sulfatase (AS) and neutrophil elastase (NE) for GCF collected from 6 samplings of 6 standard gingival sites in 11 young adult subjects over a 6-week period. Attachment loss (3 mm) was noted at only 1 of the 66 sites. The mean gingival index of the 11 subjects ranged from 0.33 to 1.67. The GCF volume and activity/sample of LDH and AS but not NE differed among subjects. However, differences among subjects were not found when the GCF enzyme activities were expressed as activity/microliter GCF. GCF volume and LDH, AS and NE activity all differed among the 6 sites when the activities were expressed as either quantity/sample or microliter GCF. These data show that differences among sites must be carefully considered in evaluation of GCF data. Fluid volume and LDH, AS and NE activity all varied from sampling to sampling. However, differences among sites were retained throughout the experimental period.     

牙周非手术治疗对重度慢性牙周炎患者龈沟液中C反应蛋白的影响

目的检测牙周基础治疗对慢性牙周炎患者龈沟液中C反应蛋白(CRP)的影响,为牙周病活动期诊断及判断牙周治疗的效果提供一定的客观依据。方法治疗前及治疗后1、3、6、12个月,用滤纸条收集30例重度慢性牙周炎患者的60个重度牙周炎牙位(探诊深度PD≥6 mm)和60个轻度牙周炎牙位(PD≤4 mm)的龈沟液并称重,用酶联免疫吸附测定法(ELISA)测定CRP的含量并记录牙周临床指标,15例牙周健康者的30个健康牙位作为对照。结果深牙周袋牙位的CRP在龈沟液中的浓度((968.06±360.54)pg/m L)显著高于浅牙周袋牙位((291.65±65.62)pg/m L),且疾病牙位的CRP浓度均显著高于健康牙位((33.47±24.53)pg/m L),龈沟液中CRP浓度与探诊深度(r=0.825,P<0.05)、附着丧失(r=0.833,P<0.05)、菌斑指数(r=0.741,P<0.05)呈正相关关系。同时,牙周基础治疗后沟液中CRP浓度明显降低,并且与口腔卫生情况有关。结论龈沟液中CRP浓度与牙周破坏程度有关,非手术治疗后龈沟液中CRP浓度下降。     

Cigarette smoking, salivary/gingival crevicular fluid cotinine and periodontal status. A 10-year longitudinal study

BACKGROUND, AIMS: The primary purpose of this study was to determine the association of salivary and gingival crevicular fluid (GCF) cotinine levels with periodontal disease status in smokers and non-smokers. METHODS: 147 male smokers and 30 male non-smokers were included in the current longitudinal study. The 177 individuals were part of a group of 200 subjects (89%) seen 10 years previously for a baseline survey. Oral hygiene indices, probing depth and attachment loss were recorded. Salivary and GCF cotinine levels of 58 smokers were determined by means of ELISA. RESULTS: Results indicated that no significant difference was found in subjects who smoked, when compared to subjects who did not smoke with respect to plaque accumulation and calculus deposits. Smokers, however, had fewer gingival bleeding sites. Cigarette smoking was associated with a greater increase in probing depth and attachment loss, as well as greater tooth loss at an earlier age. There was greater tooth loss in smokers than non-smokers (p < 0.001). 11 smokers became edentulous, while only 1 non-smoker lost all his teeth within 10 years. The degree of periodontal tissue breakdown was different in each age group with greater periodontal deterioration as age increased. All smokers had detectable salivary and GCF cotinine. Mean GCF cotinine was about 4x higher than mean salivary cotinine levels. Individuals who smoked > or = 20 pack years when compared to <20 pack years, had significantly higher saliva and GCF cotinine levels (p < or = 0.05). CONCLUSION: Neither salivary cotinine nor GCF cotinine was significantly correlated with probing depth, attachment loss and tooth loss (p > 0.05).     

Crevicular fluid myeloperoxidase at healthy, gingivitis and periodontitis sites

The volume and myeloperoxidase (MPO) activity of gingival crevicular fluid (GCF) collected with filter paper strips for 30 s from the sulcus of healthy, gingivitis and periodontitis sites of Chinese subjects were measured. MPO/site and MPO/microliter GCF were both greater at gingivitis and periodontitis sites than at healthy sites. Enzyme activity was similar at the 2 categories of diseased sites. Mean GCF volume and MPO activity were calculated for all samples from healthy, gingivitis and periodontitis sites with GI 0, 1, 2 and 0 + 1. GCF volume, MPO/site and MPO/microliter GCF all were greater at GI 2 than GI 0 or 0 + 1. These data indicate that increased GCF MPO previously observed at periodontitis sites is not specific to such sites. Rather increased GCF MPO likely occurs when additional polymorphonuclear leukocytes enter the sulcus as a result of gingival inflammation. A second sample was obtained from 22 sites 4 weeks after the initial collection. These samples were collected for 5 s rather than 30 s. The GCF volume, MPO/site and MPO/microliters GCF were each greater in samples collected for 30 s rather than 5 s. Correlation coefficients showed that the amount of GCF and MPO activity of the fluid collected for 5 s and 30 s was dependent upon the site even though the 5-s and 30-s samples were collected 4 weeks apart.     

Gingival crevicular fluid levels of aspartate amino transferase, sulfide ions and N-benzoyl-DL-arginine-2-naphthylamide in diabetic patients with chronic periodontitis

BACKGROUND: The aim of this study is to analyze the correlations between plaque index (PlI), gingival index (GI), probable pocket depth (PPD), clinical attachment level (CAL), aspartate aminotransferase (AST), N-benzoyl-DL-arginine-2-naphthylamide (BANA) and sulfide ion activity (SIA) of diabetic patients with chronic periodontitis with regard to disease activity detected by AST levels. MATERIAL AND METHODS: A total of 95 sites from eight diabetic patients with chronic periodontitis and 74 sites from eight systemically healthy patients with chronic periodontitis were enrolled in the study. The patients had no history of periodontal treatment or any antibiotic therapy during the last 6 months and were nonsmokers. All the sites selected for the study had a CAL of at least 2 mm. Gingival crevicular fluid volumes (GCFV) were measured in all sites. RESULTS: According to the result of AST analysis, 45 sites were AST positive and 50 were AST negative in the diabetic group and 36 sites were AST positive and 38 were AST negative in the control group. There was a significant correlation between BANA hydrolysis and PPD in both diabetic and control groups, but no correlation between PPD and AST levels. A significant correlation was observed between AST-positive sites and GI, but not between GI and BANA hydrolysis. In both groups, the correlation between SIA and BANA hydrolysis was significant, but no correlation was revealed between SIA and AST levels in either diabetic or control groups. CONCLUSION: The GCF metabolites had significant correlations with periodontally diseased sites in patients with chronic periodontitis, whether diabetic or systemically healthy, and may help to confirm clinical findings.     

A relationship between proteinase activity and clinical parameters in the treatment of periodontal disease

Abstract. The objective of this research was to determine the effectiveness of a biochemical assay which measures proteolytic enzyme activity in gingival crevicular fluid (GCF) and to relate this enzyme activity to clinical parameters traditionally utilized for periodontitis detection. A clinical trial was conducted on 8 periodontitis subjects with ≥4 sites exhibiting a loss of attachment of ≥5 mm and probing depths of ≥5 mm with bleeding on probing. On each subject, a plaque index was performed, followed by GCF sampling at those sites which exhibited a loss of attachment and probing depths. GCF was analyzed for activity against benzoyl-L-arginine-p-nitroanilide in the presence (BAPNA w/gly-gly) and the absence (BAPNA w/o gly-gly) of glycyl-glycine and against MeOSuc-Ala-Ala-Pro-Val-pNA and Suc-Ala-Ala-Pro-Phe-pNA for neutrophil serine proteinases activity (elastase and cathepstn G, respectively). Subsequently, a gingival index was performed, attachment levels and probing depths were recorded using a constant force probe with bleeding on probing being noted. A split-mouth design was employed and half mouths were randomly assigned to the following treatment groups: group A, half of the mouth received scaling/root planing and polishing: group B, half of the mouth received no treatment (control). Subjects were treated, then instructed on toothbrushing and interdental cleaning. After 4 weeks, subjects returned to receive a plaque index; GCF sampling, gingival index, attachment levels, probing depths and bleeding on probing as described above. Using a patred Student t-test, the findings suggest that BAPNA w/gly-gly was significantly less in treatment sites than in non-treated control sites (p=0.05). No such correlation was found for other activities, including neutrophil serine proteinases which were shown to occur in GCF in free, proteolytically active forms. In addition, significant treatment effects were detected for probing depths (p= 0.03) which reduced by 1.3 mm and attachment levels (p=0.02) which gained 0.7 mm. The reduction of P. gingivalis from treated periodontitis sites as detected by a significant decrease in BAPNA w/gly-gly may prove to be a valuable marker for periodontal disease activity.     

Phylloquinone in gingival crevicular fluid in adult periodontitis

Abstract. Phylloquinone is a lipid soluble vitamin which is an absolute growth requirement for black-pigmented anaerobes, many of which are implicated in the aetiology of periodontal diseases. This cross-sectional study aimed to detect the levels of phylloquinone in GCF from healthy and diseased sites in subjects with adult periodontitis, in order to investigate further its potential role in the disease process. The sample consisted of eighteen patients with adult periodontitis. Periodontal probing depths, attachment levels and gingival indices were recorded from one healthy and one diseased site in each subject. GCF was sampled and the amount of phylloquinone in each sample was determined using reverse-phase high performance liquid chromatography coupled to electrochemical detection. The mean amount of phylloquinone in accumulated GCF from diseased sites was 406 pg/site and 80 pg/site from healthy sites ( p =0.013). When the amounts of phylloquinone in GCF were expressed as concentrations the values were 228 ng/ml and 3350 ng/ml for diseased and healthy sites respectively ( p =0.084). These findings suggest the levels of phylloquinone in GCF differs in periodontal health and disease in subjects with adult periodontitis. The total phylloquinone at diseased sites may provide the nutritional requirements favouring the growth of black-pigmented anaerobes.     

Neutrophil elastase in crevicular fluid: comparison of a middle-aged general population with healthy and periodontitis groups

Abstract Neutrophil elastase (NE) was measured in crevicular fluid (GCF) collected from 3 subject groups. GCF was harvested at a single visit of subjects with periodontal health (n=21) and with periodontitis (n=28). Samples were obtained from 132 middle-aged, middle-class health conscious patients of a health maintenance organization (HMO) at baseline and 1 year later. GCF NE was higher in periodontitis than in health. Mean GCF NE of HMO subjects was much closer to health than to periodontitis. Few members of the HMO population had enzyme levels typical of periodontitis. Subjects and sites of the HMO population were segregated into 3 categories based on enzyme levels of the healthy and periodontitis subjects. Most HMO subjects and sites were in the activity category corresponding to healthy subjects. Only a small portion were in the activity category common in periodontitis. Enzyme levels in the highest activity category at both samplings were infrequent. High enzyme levels in the HMO population were not associated with attachment loss. Thus, assay of GCF NB provided little evidence of disease in a middle-aged, middle-class health conscious general population. This finding confirms an analysis of epidemiological surveys which concluded that a population such as studied here would not benefit from periodontal diagnostic testing.     

Identification of the osteoprotegerin/receptor activator of nuclear factor-kappa B ligand system in gingival crevicular fluid and tissue of patients with chronic periodontitis

BACKGROUND AND OBJECTIVE: Recent findings have suggested that osteoclastogenesis is directly regulated by receptor activator of nuclear factor-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG). However, no studies have described interactions of OPG/RANKL and the gp130 cytokine family in periodontal disease. This study aimed to identify and quantify OPG/RANKL in the gingival crevicular fluid (GCF) and connective tissue of patients with periodontitis, and to clarify possible correlations with disease severity and interleukin-6 (IL-6) cytokines. MATERIAL AND METHODS: Ninety-five sites in 20 patients with generalized chronic periodontitis were divided into four groups by site based on probing depth (PD) and bleeding on probing (BOP). In periodontitis patients, GCF was obtained using sterile paper strips from clinically healthy sites (PD 6 mm with BOP, n = 27). Fourteen clinically healthy sites from four periodontally healthy individuals were used as the control group. The levels of OPG, RANKL and two gp130 cytokines - IL-6 and oncostatin M (OSM) - in the GCF were determined by an enzyme-linked immunosorbent assay (ELISA) and are expressed as total amounts (pg/site). Immunohistochemical localization of OPG- and RANKL-positive cells was also performed on gingival connective tissues harvested from patients with periodontitis (inflammatory group, n = 8 biopsies) and from non-diseased individuals (healthy group, n = 8 biopsies). RESULTS: GCF RANKL, but not OPG, was elevated in diseased sites of patients with periodontitis. However, the expressions of OPG and RANKL showed no correlation with disease severity (r = 0.174 and 0.056, respectively), but the content of RANKL in the GCF was significantly positively correlated with those of IL-6 (r = 0.207) and OSM (r = 0.231) (p < 0.01). Immunohistochemical staining showed that RANKL-positive cells were significantly distributed in the inflammatory connective tissue zone of diseased gingiva, compared with those of samples from non-diseased persons (p < 0.01). However, few OPG-positive cells were found in connective tissue zones of either the diseased gingiva or healthy biopsies. CONCLUSION: These findings imply that in this cross-sectional study of GCF, RANKL, IL-6 and OSM were all prominent in periodontitis sites, whereas OPG was inconsistently found in a few samples of diseased sites but was undetectable in any of the control sites. The results also imply that the expression of RANKL was positively correlated with IL-6 and OSM in the GCF.     

Granulocyte elastase in gingival crevicular fluid: improved monitoring of the site-specific response to treatment in patients with destructive periodontitis

Abstract In 13 patients with severe destructive periodontitis. the response to periodontal therapy was estimated by granulocyte elastase level in gingival crevicular fluid (GCF). 62 sites were classified according to changes of probing depths (PD) and quantitative bone height (BH%) before and after 5–year regular maintenance treatment: (i) 17 consistently healthy sites with no changes of PD and BH%; (ii) 6 initially healthy sites with deterioration in PD and BH%; (iii) 14 diseased sites with improvement in PD and BH%; (iv) 25 diseased sites with no improvement in PD and BH%. GCF was collected by an intracrevicular washing system. The released elastase in the supernatants (EA-S) and the cell-bound elastase in the pellets (EA-P) were determined with a low molecular weight substrate specific for granulocyte elastase. The ratio of EA-S and EA-P (S/P-ratio) was used as a relative measure of elastase released by the granulocytes present. The sites classified as diseased with no improvement or initially healthy but deteriorating, had significantly higher EA-S, EA-P and S/P-ratios than the consistently healthy sites or diseased but improving sites (p < 0.01). Both EA-S and S/P-ratio showed strongly positive correlations with the current levels of gingival inflammation and periodontal destruction (p < 0.001). The present study suggests that increased elastase level is associated with disease progression, and may be used to monitor the response to longitudinal maintenance therapy.