Crevicular fluid endothelin-1 levels in periodontal health and disease

Background and Objective:  Endothelin-1 is a 21-amino-acid peptide with multifunctional regulation. Initial research indicated that endothelin-1 levels in the gingival crevicular fluid from patients with chronic periodontitis were higher than those in the gingival crevicular fluid from healthy subjects. The aim of the present study was to assess the relationship between the clinical parameters and the concentrations of endothelin-1 within the gingival crevicular fluid from inflamed gingiva and periodontitis sites and, subsequently, after the treatment of periodontitis sites.
Material and Methods:  A total of 60 subjects were divided into three groups – healthy (group I), gingivitis (group II) and chronic periodontitis (group III) – based on gingival index, pocket probing depth and clinical attachment loss. A fourth group consisted of 20 subjects from group III, 6–8 wk after treatment (i.e. scaling and root planing). Gingival crevicular fluid samples collected from each patient were quantified for endothelin-1 using an enzymatic immunometric assay.
Results:  Endothelin-1 was not detected in any sample from any of the study groups.
Conclusion:  The results showed that all the gingival crevicular fluid samples were negative for the endothelin-1 molecule. Therefore, endothelin-1 cannot be considered as a potential biomarker of periodontal disease progression.     

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.     

Effect of scaling and root planing on interleukin-1β, interleukin-8 and MMP-8 levels in gingival crevicular fluid from chronic periodontitis patients

Konopka ?, Pietrzak A, Brzezińska‐B?aszczyk E. Effect of scaling and root planing on interleukin‐1β, interleukin‐8 and MMP‐8 levels in gingival crevicular fluid from chronic periodontitis patients. J Periodont Res 2012; 47: 681–688. © 2012 John Wiley & Sons A/S Background and Objective: There are few data concerning the effect of scaling and root planing on the levels of immune and inflammatory mediators in gingival crevicular fluid from patients with chronic periodontitis. Therefore, in this study the influence of scaling and root planing was determined on amounts of interleukin (IL)‐1β, IL‐8 and MMP‐8 in gingival crevicular fluid from patients with chronic periodontitis, in relation to clinical parameters. Material and Methods: A total of 51 patients were enrolled in this study. The study population consisted of 30 patients with generalized advanced chronic periodontitis, while 21 periodontally healthy subjects were recruited for the control group. The clinical parameters included approximal plaque index, gingival index, pocket depth and clinical attachment loss. The amounts of IL‐1β, IL‐8 and MMP‐8 in gingival crevicular fluid were measured by ELISA. Periodontal parameters as well as gingival crevicular fluid humoral factor amounts were evaluated in the control group and in chronic periodontitis patients at baseline and at 1 and 4 wk after scaling and root planing treatment. Results: At baseline, there were significant differences between control subjects and chronic periodontitis patients in terms of clinical attachment loss, pocket depth, gingival index (p < 0.001) and approximal plaque index (p < 0.01). The amounts of IL‐1β, MMP‐8 (p < 0.001) and IL‐8 (p < 0.01) in gingival crevicular fluid were significantly lower in healthy subjects than in chronic periodontitis patients. Scaling and root planing led to improvement in all examined clinical parameters, apart from clinical attachment loss. Periodontal treatment also resulted in a significant decrease in the amounts of IL‐1β, IL‐8 and MMP‐8 in comparison to baseline, especially 4 wk after scaling and root planing (p < 0.001); however, the amounts of these humoral factors were still higher than those in control group. Conclusion: Our observations indicated that short‐term nonsurgical therapy resulted in a significant improvement in periodontal indices and in a marked decrease of IL‐1β, IL‐8 and MMP‐8 gingival crevicular fluid levels. Nevertheless, no significant correlations were found between clinical parameters and amounts of humoral factors after therapy.     

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.     

Progressive periodontal disease has a simultaneous incremental elevation of gingival crevicular fluid and serum CRP levels

Aim: Increased C‐reactive protein levels have been found in all active inflammations, including periodontitis. This study aims to assess the C‐reactive protein levels in periodontal disease progression. Methods: Forty‐five patients were divided into the following three groups (n = 15) based on gingival index, probing pocket depth, and clinical attachment level: healthy (group I), gingivitis (group II), and chronic periodontitis (group III). Gingival crevicular fluid and serum samples were quantified for C‐reactive protein using enzyme‐linked immunosorbent assay. Results: The mean C‐reactive protein concentration in gingival crevicular fluid and serum was found to be highest in group III (1233.33 ng/mL for gingival crevicular fluid, 5483.33 ng/mL for serum), and least in group I (60 ng/mL and 413 ng/mL for gingival crevicular fluid and serum, respectively) The mean C‐reactive protein concentration in group II (453.33 ng/mL for gingival crevicular fluid and 3565.33 ng/mL for serum) was found to be intermediate. Conclusions: C‐reactive protein levels in gingival crevicular fluid and serum increased proportionately with the severity of periodontal disease. They correlated positively with clinical parameters, including gingival index, probing pocket depth, and clinical attachment level. Thus, it can be considered as a periodontal inflammatory biomarker and deserves further consideration.     

龈沟冲洗液的蛋白质含量测定

本研究采用龈沟冲洗法检测牙周健康部位和疾病部位的龈沟液蛋白质含量，旨在分析蛋白捻民牙周病变的关系。结果表明 周炎组的龈沟因吸液量及总蛋白含量显著高于健康、龈沟炎组，健康与龈炎组的回吸液量几乎无差别。龈沟回吸液量和蛋白号含量均与临床指标－－出血指数、菌班指数、探诊深度、附着丧失显著相关，其中与探诊深度的相关性较大，提示龈沟洗法所得冲洗液量受牙周袋深度的影响，回吸液中蛋白含量随着临床炎症加重而升高，说     

Gingival crevicular fluid and plasma levels of neuropeptide Substance-P in periodontal health, disease and after nonsurgical therapy

Background and Objective: The level of Substance‐P in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and levels of Substance‐P in the gingival crevicular fluid from inflamed gingiva, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the Substance‐P levels of plasma. Material and Methods: Thirty, age‐ and gender‐matched subjects were divided into three groups (healthy, gingivitis and chronic periodontitis) based on modified gingival index scores and clinical attachment loss. A fourth group consisted of 10 subjects from the periodontitis group, 6–8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for Substance‐P using an enzyme immunoassay. Results: The mean concentration of Substance‐P, both in gingival crevicular fluid and plasma, was observed to be highest in the periodontitis group (45.13 pg/mL in gingival crevicular fluid and 67.8 pg/mL in plasma) and lowest in the healthy group (6.07 pg/mL in gingival crevicular fluid and below the detection level in plasma). The mean Substance‐P concentration in the gingivitis group (11.42 pg/mL in gingival crevicular fluid and 38.8 pg/mL in plasma) and in the after‐treatment group (7.58 pg/mL in gingival crevicular fluid and 39.7 pg/mL in plasma) lay between the highest and lowest values. In all groups the gingival crevicular fluid levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. Conclusion: Substance‐P levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of Substance‐P levels. Gingival crevicular fluid and plasma Substance‐P levels showed a positive correlation in all of the groups.     

Levels of specific immunoglobulin G to the forsythia detaching factor of Tannerella forsythia in gingival crevicular fluid are related to the periodontal status

Onishi H, Arakawa S, Nakajima T, Izumi Y. Levels of specific immunoglobulin G to the forsythia detaching factor of Tannerella forsythia in gingival crevicular fluid are related to the periodontal status. J Periodont Res 2010; 45: 672–680. © 2010 John Wiley & Sons A/S Background and Objective: Forsythia detaching factor (FDF) is a putative virulence factor of Tannerella forsythia that induces detachment of adherent cells and interleukin‐8 production in human fibroblasts. The objective of the present study was to clarify the relationship between anti‐FDF IgG levels in gingival crevicular fluid and the clinical status in patients with periodontitis and in healthy subjects. Material and Methods: Gingival crevicular fluid and subgingival plaque samples were obtained from both the diseased and healthy sites of 37 patients with periodontitis and from 30 healthy subjects. Anti‐FDF IgG levels were evaluated, and both the fdf gene and T. forsythia 16S ribosomal RNA (rRNA) were detected using the PCR. Results: Anti‐FDF IgG levels (of both diseased and healthy sites) of patients with periodontitis were significantly higher than those of healthy subjects. Among the patients with periodontitis, anti‐FDF IgG levels of healthy sites were significantly higher than those of diseased sites and the levels showed negative correlations with probing pocket depth and clinical attachment level. Among the patients with periodontitis, T. forsythia 16S rRNA was detected in 18 of 37 diseased sites and in 5 of 29 healthy sites, and the fdf gene was detected in 19 of 37 diseased sites and in 7 of 29 healthy sites. By contrast, no healthy subjects were positive for T. forsythia 16S rRNA or the fdf gene. Conclusion: These data suggest that anti‐FDF IgG levels in gingival crevicular fluid are related to the periodontal status.     

The electrophoretic detection of acidic glycosaminoglycans in human gingival sulcus fluid

Fluid collected from non-inflamed sites contained only hyaluronic acid, whereas fluid collected from severely inflamed sites possessed additional components corresponding in electrophoretic mobility to dermatan sulphate and chondroitin-4-sulphate. The results imply that the source of gingival fluid glycosaminoglycans is the associated periodontal tissue and that the fluid may well be a vehicle for the transport and subsequent detection of glycosaminoglycans in the tooth integuments.     

慢性牙周炎患者牙周治疗前后龈沟液中白介素-10水平的变化

目的探讨慢性牙周炎患者牙周治疗前后龈沟液中抗炎性细胞因子白介素(IL)-10水平的变化。方法采集12例慢性牙周炎患者的12个健康牙位和36个炎症牙位于治疗前及治疗后6、122、4周的龈沟液,用酶联免疫吸附分析法(ELISA)检测龈沟液中IL-10的浓度。另外,分别记录治疗前、后的探诊深度(PD)、临床附着丧失(CAL)、牙龈指数(GI)和菌斑指数(PlI)。结果IL-10浓度在健康牙位明显高于炎症牙位(P<0.01),且于牙周治疗后明显升高。IL-10浓度与探诊深度(PD)、临床附着丧失(CAL)呈负相关(P<0.05)。结论IL-10浓度与牙周组织破坏程度呈负相关,在牙周炎中起抗炎作用。     

Full-mouth profile of active MMP-8 in periodontitis patients

Kraft‐Neumärker M, Lorenz K, Koch R, Hoffmann T, Mäntylä P, Sorsa T, Netuschil L. Full‐mouth profile of active MMP‐8 in periodontitis patients. J Periodont Res 2012; 47: 121–128. © 2011 John Wiley & Sons A/S Background and Objective: MMP‐8 in gingival crevicular fluid is considered as a protease with high destructive potential because of its ability to degrade collagen in periodontitis‐affected patients. The aim of this study was to investigate whether there was a relationship between clinical diagnostic parameters and the concentration of active MMP‐8 (aMMP‐8) in gingival crevicular fluid in a site‐level full‐mouth analysis. Based on these data, the prognostic value of aMMP‐8 levels in relation to pocket depth may be evaluated. Material and Methods: Clinical measurements of pocket depth, bleeding on probing (BOP), plaque index (PlI) and gingival index (GI), as well as samples of gingival crevicular fluid, were obtained from four sites of each tooth of nine healthy female patients with chronic generalized periodontitis. The aMMP‐8 concentration in gingival crevicular fluid was quantified by ELISA using specific monoclonal antibodies. Multiple linear regression models for the single measures of aMMP‐8 and pocket depth were calculated with GI and BOP as additional variables. Results: Between 92 and 112 recordings were obtained for each parameter in each patient. Mean values of between 31.5 and 88.8% were calculated for pocket depths of ≥ 4 mm. Mean pocket depths ranged from 3.11 to 4.73 mm, the mean BOP values ranged from 34.0 to 96.7% and the mean full‐mouth gingival crevicular fluid aMMP‐8 concentration ranged from 3.2 to 23.7 ng/mL. Conclusion: In this sample of female periodontitis patients, a broad range of intra‐individual and interindividual aMMP‐8 values was found. Although the explained variance was rather weak, a statistically significant relationship between aMMP‐8 and pocket depth was proven.     

Analysis of proteins in human gingival crevicular fluid by mass spectrometry

Kido J, Bando M, Hiroshima Y, Iwasaka H, Yamada K, Ohgami N, Nambu T, Kataoka M, Yamamoto T, Shinohara Y, Sagawa I, Nagata T. Analysis of proteins in human gingival crevicular fluid by mass spectrometry. J Periodont Res 2012; 47: 488–499. © 2012 John Wiley & Sons A/S Background and Objective: Gingival crevicular fluid is a bodily fluid transuded from periodontal tissues into the gingival crevice and periodontal pocket, and contains many species of components. Proteins in gingival crevicular fluid have been studied as markers for periodontal diseases. Mass spectrometric analysis is used for the analyses of proteins, lipids, saccharides and metals, and expected as an approach for disease diagnosis. For better analysis of the protein components in gingival crevicular fluid, we investigated proteins in gingival crevicular fluid samples from the healthy gingival crevice and periodontal pocket using mass spectrometry. Material and Methods: Gingival crevicular fluid samples were collected from subjects who gave their informed consent and were periodontally healthy or had diseased pockets. These samples were electrophoretically separated, and each fraction on the gels was analysed by nano liquid chromatography coupled with tandem mass spectrometry. Antimicrobial peptides detected in gingival crevicular fluid were confirmed by western blotting. Results: One hundred and four proteins were detected in gingival crevicular fluid samples from both healthy sites and sites of periodontitis; 64 proteins were contained only in gingival crevicular fluid from healthy sites and 63 proteins were observed only in gingival crevicular fluid from periodontitis sites. These proteins were blood‐, cytoskeleton‐, immunity‐, inflammation‐ and lipid‐related proteins and enzymes. Some proteins, including ceruloplasmin, glycogen phosphorylase, glutathione S‐transferase, phosphoglycerate mutase, psoriasin, S100A11 and resistin, were identified for the first time in gingival crevicular fluid. Antimicrobial peptides, such as lactoferrin, α1‐antitrypsin, lipocalin, S100A7, S100A8, S100A9 and cathelicidin, were observed by mass spectrometry and western blotting. Conclusion: Multiple protein components in gingival crevicular fluid were analysed at the same time using mass spectrometry, and this approach may be useful for the diagnosis of periodontal diseases.     

Plasma and crevicular fluid osteopontin levels in periodontal health and disease

BACKGROUND AND OBJECTIVE: The level of osteopontin in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and osteopontin levels of the gingival crevicular fluid from inflamed gingivae, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the osteopontin levels of the plasma. MATERIAL AND METHODS: Thirty, gender-matched subjects were divided into three groups--healthy, gingivitis and chronic periodontitis--based on modified gingival index scores and clinical attachment loss. The fourth group consisted of 10 subjects in the periodontitis group, 6-8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for osteopontin using an enzyme immunoassay. RESULTS: The highest mean gingival crevicular fluid and plasma osteopontin concentrations were observed in the periodontitis group (1575.01 and 1273.21 ng/mL, respectively) and the lowest in the healthy group (1194.80 and 476.35 ng/mL, respectively). After treatment of the periodontitis group, the level of osteopontin decreased to 1416.15 in gingival crevicular fluid and to 1051.68 ng/mL in plasma. In all groups the gingival crevicular fluid osteopontin levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. CONCLUSION: Osteopontin levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of osteopontin levels. Gingival crevicular fluid and plasma osteopontin levels showed a positive correlation in all of the groups.     

龈沟液中神经肽P物质与慢性牙周炎关系临床研究

目的:探讨龈沟液中神经肽P物质(SP)与慢性牙周炎发生发展的关系.方法:研究组48 例,对照组45 例,研究组口内各选1 个牙周炎牙位、对照组各选1 个健康牙位收集龈沟液样本,采用放射免疫分析技术分别测定2 组龈沟液SP含量;记录牙龈指数(GI)、牙周袋探诊深度(PD)、附着丧失(AL)、牙槽骨吸收(ABL)情况,并进行与SP的相关性分析.结果:研究组SP含量显著高于对照组(t＝9.92,P<0.01);轻度牙周炎SP含量及牙周临床指标与对照组相比,均有显著差异(P<0.01);轻度牙周炎龈沟液SP含量与GI无显著相关性,但与PD、AL、ABL之间,以及中、重度牙周炎龈沟液SP含量与GI之间,均呈显著性相关(P＜0.05);中、重度牙周炎SP含量与PD、AL、ABL之间均呈非常显著性相关(P<0.01).结论:龈沟液中SP含量与慢性牙周炎的发生发展密切相关,可以反映牙周组织的破坏程度;测定患者龈沟液中SP含量,对于判断病情、指导治疗具有重要意义.     

Lipid peroxidation: a possible role in the induction and progression of chronic periodontitis

OBJECTIVES: Reactive oxygen species (ROS) are implicated in the destruction of the periodontium during inflammatory periodontal diseases. The imbalance in oxidant/antioxidant activity may be a key factor in the damaging effects of ROS. This study aimed to determine the lipid peroxidation levels in gingival crevicular fluid and saliva, and glutathione (GSH) and glutathione peroxidase (GPx) in saliva in patients with chronic periodontitis. METHODS: Gingival crevicular fluid and saliva were collected from 13 patients and 9 healthy control subjects during the preliminary study, and from 21 patients during the subsequent study. Lipid peroxidation level, GSH level and GPx activity were determined by spectrophotometric assay. RESULTS: The preliminary study found that when comparing patients to healthy controls, the gingival crevicular fluid samples produced the following results, respectively: higher lipid peroxidation concentration (microm) (by sites: 167.55 vs. 53.71, p < 0.0001; by subjects: 151.99 vs. 50.66, p < 0.005) and total amount (pmol) (by sites: 93.02 vs. 8.47, p < 0.0001, by subjects: 80.44 vs. 7.84, p < 0.0005). In saliva samples, lower GSH concentration (microm) (373.04 vs. 606.67, p < 0.05), higher lipid peroxidation concentration (microm) (0.66 vs. 0.13, p < 0.0005), and no difference in GPx activity were found in patients than in those of healthy controls. The subsequent study showed statistically significant (p < 0.05) improvement of clinical periodontal parameters (plaque index, gingival index, probing attachment level, probing pocket depth and gingival crevicular fluid volume), decreases in gingival crevicular fluid lipid peroxidation levels (concentration and total amount) at the sites after the completion of phase 1 periodontal treatment. Similarly, the periodontal treatment resulted in a significant decrease of lipid peroxidation concentrations (p < 0.05), increase in GSH concentration (p < 0.001), and no change in GPx activity in saliva samples. CONCLUSION: The increased levels of lipid peroxidation may play a role in the inflammation and destruction of the periodontium in periodontitis.     

Neutrophil elastase activity, levels of prostaglandin E2, and matrix metalloproteinase-8 in refractory periodontitis sites in smokers and non-smokers.

The study was aimed to determine elastase activity, levels of prostaglandin E2 (PGE2), and matrix metalloproteinase-8 (MMP-8) in gingival crevicular fluid (GCF) in 20 smokers and 20 non-smokers, mean age 47.4 (+/-2.9 SD) years with refractory periodontal diseases. GCF was collected with intracrevicular washing from four sites in each subject. Clinical assessments, included gingival index, probing depth, clinical attachment level, bleeding on probing, bone height, and plaque accumulation. Smokers had a significantly higher percentage of the gingival margin covered by plaque (P%Im), higher number of sites with probing pocket depth > or = 5 mm, higher mean values of probing pocket depth and probing attachment level (P< 0.01). Smokers had significantly higher mean levels of neutrophil elastase activity (P< 0.01) in the supernatants than non-smokers did. In sites with matching pocket depths, neutrophil elastase activity was significantly higher in smokers (P< 0.001) than in non-smokers. In sites with high levels of MMP-8 the PGE2 levels were significantly (P< 0.001) higher compared to sites with low levels in smokers as well as in non-smokers. A significant correlation was found between probing pocket depth and levels of MMP-8 (P< 0.001) and in non-smokers between probing pocket depth and levels of PGE2 (P< 0.05).     

Variations in crevicular fluid elastase levels in periodontitis patients on long-term maintenance

Granulocyte elastase was determined in the gingival crevicular fluid (GCF) of 18 periodontitis patients. They initially had similar severity of disease but had responded differently to 5-yr maintenance, 13 responders and 5 non-responders. A total of 102 sites were investigated and categorized as: i) consistently healthy, ii) healthy after treatment, iii) gingivitis, and iiii) periodontitis, according to clinical criteria. GCF elastase activity was determined with a granulocyte-specific substrate. The sites from non-responders had consistently higher elastase levels than the corresponding category of sites from responders, despite similar gingival inflammation and periodontal destruction, with the exception of consistently healthy sites. Within the non-responders, the periodontitis sites had higher elastase levels than the gingivitis sites commensurate with probing depth, while no difference existed between gingivitis sites and sites healthy after treatment, despite a difference in probing depth. In contrast, in the responders similar elastase levels were found at the periodontitis sites and gingivitis sites despite difference in probing depth, while both diseased sites had higher elastase levels than the sites healthy after treatment, commensurate with probing depth. This study suggests that increased granulocyte-specific elastase levels in GCF may serve as a diagnostic marker for refractory periodontitis patients.     

The subgingival microflora and gingival crevicular fluid cytokines in refractory periodontitis

Abstract Refractory periodontitis manifests as a rapid, unrelenting, progressive loss of attachment despite the type and frequency of therapy. This study examined possible relationships between cytokine levels in gingival crevicular fluid (GCF), occurrence of specific periodontopathic microflora. and disease activity in patients with refractory periodontitis. Refractory periodontitis patients (7 male and 3 female) were selected on the basis of history and longitudinal clinical observations. In each patient. 2 teeth with pocket depths greater than 6 mm were selected and individual acrylic stents were fabricated with reference grooves for each site. The sites were examined at both baseline and 3 months later. The pattern and amount of alveolar bone resorption were assayed by quantitative digital subtraction radiography. Pocket depth and attachment loss were measured with a Florida Probe. The gingival index was measured at 4 sites around each sample tooth. Sites were divided into active sites (2.1 mm loss of attachment in 3 months) or inactive sites (2.0 mm loss of attachment in 3 months). The distribution and prevalence of the predominant microflora in active and inactive sites were compared using anaerobic culture and indirect immunofluorescence. Interleukin-1β, 2, 4, 6 and tumor necrosis factor-α (TNF-α) levels in gingival crevicular fluid (GCF) were quantified by ELISA. Prevotella intermedia and Eikenella corrodens significantly decreased in inactive sites but remained the same in active sites after 3 months. The active sites revealed significantly higher GCF levels of IL-2 and IL-6 than inactive sites at both baseline and at 3 months. IL-1β was also significantly greater in active sites than in inactive sites at 3 months. Alveolar bone loss in active sites correlated with increased GCF levels of IL-1β and 1L-β. These results suggest that GCF levels of IL-1β, IL-2 and IL-6 and P. intermedia and E. corrodens in subgingival plaque may serve as possible indicators of disease activity in refractory periodontitis.     

Relationship between gingival crevicular fluid levels of aspartate aminotransferase and active tissue destruction in treated chronic periodontitis patients

Data from several sources demonstrate that disease-active and disease-inactive periodontal pockets exist, and that disease progression occurs in bursts of activity. Currently used diagnostic procedures do not distinguish between disease-active and disease-inactive sites at any given point in time. We report the results of studies aimed at determining whether levels of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) are associated with disease activity as assessed by the level of gingival inflammation and probing attachment loss. 25 previously treated periodontitis patients participating in a quarterly recall maintenance program, who had experienced recurrent periodontal deterioration, served as experimental subjects. Patients were evaluated at 3-month intervals for 2 years. Values for plaque index, gingival index, and probing attachment level were recorded, and 30-second samples of gingival fluid harvested from the mesiobuccal aspect of the 4 first molars and the distal of the 4 lateral incisors. GCF volume was measured using a Periotron 6000, and AST activity was measured by a standard method. Sites were ranked in a hierarchy based on the degree of certainty of attachment loss as well as the severity of gingival inflammation, and the relationship of the values to AST levels was determined. Three models were used to analyze the resulting data, and all led to the same conclusion. Maximum enzyme level was significantly elevated at sites with confirmed disease activity as assessed by attachment loss, with maximum AST levels 725 units higher at these sites, on average, than at other sites (p less than 0.0001). Our data support the idea that an objective diagnostic test, based on levels of AST in GCF, that distinguishes between disease-active and disease-inactive sites may be possible.     

Detection of stable and active periodontitis sites by clinical assessment and gingival crevicular acute-phase protein levels

The aim of the present study was to investigate whether incipient periodontal disease breakdown could be associated with changes in gingival crevicular fluid (GCF) acute-phase protein levels. In addition, the potential of clinical indices to act as predictors of significant attachment level (AL) change was investigated. AL measurements were taken at baseline and 3 months using the Florida Probe stent handpiece from a total of 384 sites in 38 patients. The average standard deviation of duplicate AL measurements was 0.423. When the tolerance method was used to detect significant AL change, 3.9% of the sites lost attachment. When a less stringent criterion of AL change of ≥1 mm was used 9.9% of the sites lost attachment during the 3-month period. With the exception of probing depth, baseline clinical parameters failed to predict AL change. Fourteen active periodontitis sites that demonstrated significant attachment loss were paired to stable periodontitis sites within the same patient. The levels of four acute-phase proteins, namely α2-macroglobulin (α2-M), α1-antitrypsin (α1-AT), transferrin (TF) and lactoferrin (LF), and also albumin (Alb) were assessed in the same gingival crevicular fluid sample using sandwich ELISAs. Results were expressed either as ng/30 s and ng/μg Alb. Acute-phase protein levels in GCF failed to differentiate between active and stable periodontitis sites at baseline. In conclusion, the degree of gingival inflammation of the tissues adjacent to the crevice/pocket seems to influence the levels of protease inhibitors and iron-binding proteins in GCF to a greater extent than probing attachment loss.