Gingival crevicular fluid glycosaminoglycan levels in patients with chronic adult periodontitis

Abstract This study investigated levels of hyaluronan and chondroitin-4-sulphate in the crevicular fluid of patients with chronic adult periodontitis at diseased and healthy sites before and after treatment. The relationship between clinical diagnostic parameters and levels of glycosaminoglycans in gingival crevicular fluid were also analysed. Within each patient. 4 sites either mesial or distal and on single rooted teeth were classified as diseased or healthy using a modified gingival index, pocket depth and attachment loss. Crevicular fluid was collected from each site using glass micropipettes and analysed for glycosaminoglycan content by cellulose acetate electrophoresis. Significantly higher levels of chondroitin-4-sulphate were detected at diseased sites prior to treatment correlating with increased pocket depth or attachment levels. Following a period of treatment consisting of oral hygiene instruction and root planing, the patients were reassessed for their response to treatment by measuring the modified gingival index, pocket depth, attachment loss and levels of glycosaminoglycans. Analysis of glycosaminoglycan levels at diseased sites that demonstrated a poor response to treatment also demonstrated significantly higher levels of chondroitin-4-sulphate than those sites that responded well to treatment. Hyaluronan levels were less significantly associated with clinically succesful treatment. This study confirmed the use of the sulphated glycosaminoglycan chondroitin-4–sulphate as a potential diagnostic aid of periodontal tissue destruction; however, further longitudinal studies are required to assess their performance.     

Gingival crevicular fluid alkaline phosphatase levels in postmenopausal women: effects of phase I periodontal treatment

BACKGROUND: The aim of this study was to determine how estrogen status may possibly influence gingival crevicular fluid (GCF) alkaline phosphatase (ALP) levels in estrogen-deficient (ED) and -sufficient (ES) postmenopausal women at baseline (BL) and 1 year after periodontal phase I treatment (AT). METHODS: Thirty-six postmenopausal women on estrogen supplements (mean serum estradiol levels >30 pg/ml; estrogen sufficient) and 37 postmenopausal women not on estrogen supplements (mean serum estradiol levels <30 pg/ml; ED) were divided into two subgroups as chronic periodontitis and clinically healthy controls after clinical and radiographic examination. The ES group consisted of 19 control (ES/C) and 17 chronic periodontitis (ES/P) patients, and the ED group consisted of 20 control (ED/C) and 17 chronic periodontitis (ED/P) patients. Plaque (PI) and gingival (GI) indices, bleeding on probing (BOP), probing depths (PD), clinical attachment loss (CAL) scores, and GCF samples were recorded at BL and AT. The levels of ALP in the GCF were measured photometrically. The paired samples Student t and Wilcoxon tests were used to compare the ALP levels and clinical parameters between BL and AT. The correlation among the ALP and clinical parameters was analyzed using the Pearson correlation. RESULTS: The mean of all clinical parameters (PI, GI, BOP, PD, and CAL) was significantly (P <0.05) higher in periodontitis groups (ES/P and ED/P) than controls (ES/C and ED/C). Periodontitis groups (ES/P and ED/P) demonstrated significantly increased GCF volumes and GCF ALP levels (P <0.05) compared to controls (ES/C and ED/C). There were no significant differences in the concentrations of ALP between periodontitis and control groups (P >0.05). The BL GCF ALP total levels of the ED/P group were significantly higher than the ES/P group (P <0.05). The BL and AT serum ALP levels of the ED/P group were not significantly but were numerically higher than the ES/P group. One year after periodontal treatment, the GCF volume, GCF ALP total, and concentrations decreased significantly in both periodontitis groups (P <0.05). However, the GCF ALP levels were still numerically higher in the ED/P group. A positive statistical correlation was found between total ALP levels and PD (r = 0.621; P <0.05). CONCLUSION: These data suggest that the presence of ALP in GCF is not simply a reflection of the local inflammation state and that a patient's estrogen status may possibly influence local ALP levels in GCF.     

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.     

Gingival crevicular fluid protein content and alkaline phosphatase activity in relation to pubertal growth phase

Objective:To evaluate gingival crevicular fluid (GCF) protein content and alkaline phosphatase (ALP) activity in growing subjects in relation to stages of skeletal maturation, ie, the growth phase, as prepubertal, pubertal, and postpubertal.Subjects and Methods:Fifty healthy growing subjects (31 girls and 19 boys; age range, 7.8–17.7 years) were enrolled in this study that followed a double-blind, prospective, cross-sectional design. Collection of GCF was performed at the mesial and distal sites of both central incisors, for the maxilla and mandible. Growth phase was assessed through the cervical vertebral maturation method. GCF parameters were expressed as total protein content, total ALP activity, and normalized ALP activity.Results:The total GCF protein content was similar between the different growth phases. On the contrary, the total ALP activity showed a peak for the pubertal growth phase. The normalized GCF ALP activity was only poorly associated with growth phase. No differences were seen between the maxillary and mandibular sites, or between the sexes, for any GCF parameter.Conclusions:The total GCF protein content is not sensitive to the growth phase. However, GCF ALP activity has potential as a diagnostic aid for identification of the pubertal growth phase in individual subjects when expressed as total, but not normalized, values.     

Gingival tissue and crevicular fluid co-operation in adult periodontitis

Activated matrix metalloproteinase-3 (MMP-3) can contribute to periodontal ligament destruction in adult periodontitis. Since MMP-3 has been reported to activate proMMP-8 and -9, it was speculated that gingival tissue fibroblast-derived MMP-3 might, in periodontitis, be responsible for activation of gingival crevicular fluid (GCF) neutrophil-derived proMMP-8 and -9. Immunohistochemistry disclosed MMP-3 in gingival fibroblasts in periodontitis. Cultured gingival fibroblasts released only pro-MMP-3 when stimulated with tumor necrosis factor-alpha. However, Western blot revealed partially activated MMP-3, MMP-8, and MMP-9 in periodontitis GCF. Active MMP-8 (p < 0.05) and MMP-9 (p < 0.05) correlated with the presence of active MMP-3. It seems that resident gingival fibroblasts produce pro-MMP-3 in GCF, where it becomes activated, probably by cathepsin G or elastase released by neutrophils. Active MMP-3 then activates neutrophil-derived pro-MMP-8 and -9. Different tissue compartments/cells exert co-operative actions in mutual local MMP activation cascades.     

Gingival crevicular aspartate aminotransferase levels in periodontitis patients before and after periodontal treatment]

Gingival crevicular fluid samples were collected from 296 teeth from 40 subjects, including 19 rapidly progressive periodontitis (RPP), 8 chronic adult periodontitis (CAP), 7 marginal gingivitis (MG) and 6 healthy subjects (H). The activities of aspartate aminotransferase (AST) in each sample were tested. The results were as follows: (1) The two groups with destructive periodontal disease (RPP and CAP) had greater GCF-AST levels than that from the two non-destructive groups (MG and H). (2) The GCF-AST activities showed significant positive correlations with clinical periodontal parameters, such as probing depth, attachment loss, bleeding index and suppuration. (3) Four weeks after thorough full-mouth root planing, both clinical parameters and GCF-AST levels decreased significantly. The present study suggests that GCF-AST activity might be a sensitive and objective marker for detection of periodontal tissue destruction and inflammation.     

Gingival crevicular fluid leptin levels in periodontitis patients with long-term and heavy smoking

BACKGROUND: The aim of the present study was to evaluate gingival crevicular fluid (GCF) leptin levels and the influence of long-term and heavy smoking on GCF leptin levels in patients with chronic periodontitis. METHODS: In this study, 143 individuals were divided into three groups: non-smokers (NS), smokers (S), and control (C). Three subgroups of NS and S were grouped as follows: a) probing depth (PD) 5 mm. For each patient, PD, gingival index (GI), plaque index (PI), gingival bleeding time index (GBTI), and clinical attachment level (CAL) values were recorded. The GCF leptin levels obtained from sampling sites were determined by using enzyme-linked immunosorbent assay kits. RESULTS: The GCF leptin levels were found significantly lower in the a and b subgroups in the S group than those in the NS group (P <0.05). The inflammatory markers GI and GBTI showed significant correlations with leptin in NS (P <0.05). CONCLUSIONS: Our results suggest that higher leptin GCF levels in healthy sites in periodontitis patients may play a protective role in periodontal disease. Further studies are needed to determine the cellular origin of the leptin in the gingiva and the effect of plasma leptin levels on GCF leptin concentrations.     

Gingival crevicular fluid fibronectin degradation in periodontal health and disease

The molecular forms of fibronectin (FN) in gingival crevicular fluid of five subjects with at least two sites exhibiting clinical signs of inflammation and pockets of at least 4 mm (test group) and five subjects with clinically healthy periodontium (control group) were investigated. Samples were collected with standard filter paper strips. In the test group samples from both diseased and healthy sites were collected. After collection the test group received one episode of periodontal treatment (scaling and root planning). The sampling and clinical recording were repeated for the diseased sites after about 2 wk. The crevicular fluid FN was analyzed using sodium dodecyl sulphate gel electrophoresis followed by western blotting with polyclonal antibodies against FN. Both intact FN and FN fragments were found in all samples. A larger proportion of FN was in degraded form in the diseased sites than in the healthy or the treated sites. FN was also degraded into smaller peptide fragments in the diseased than in the treated sites. These results suggest that crevicular fluid FN is partially degraded both in periodontal health and disease and that the degree of degradation of FN increases with periodontal inflammation and decreases with periodontal treatment.     

Gingival crevicular fluid plasmin activity in different clinical conditions and after periodontal treatment.

Plasmin is an important enzyme in the process of wound healing and it has been suggested that it plays a role in the pathogenesis and healing of periodontal diseases. In this study, plasmin activity in crevicular fluid (CF) was measured in different clinical conditions and after periodontal treatment. 152 CF samples were collected with paper strips from 12 subjects. Five other subjects received a single periodontal treatment after the first sample collection and the post-treatment samples were collected 2, 5, 10, 20 and 40 days thereafter. The individual strips were placed immediately on plasminogen-free fibrin plates or in plastic vials containing sterile saline. The vials were agitated and 20 microliters of each solution was pipetted onto fibrin plates. The remaining solution was used in protein analysis. The plates were incubated for 24 hrs at 37 degrees C and the activity of plasmin determined by weighing the fibrin liquified by the samples. The variation of CF plasmin activity in different pockets within individuals was larger than between the subjects. There was a weak positive correlation between CF plasmin activity and the amount of plaque, bleeding tendency, pocket depth and bone loss by x-ray. However, plasmin concentration (units of plasmin activity per CF volume) did not correlate with clinical parameters. A weak negative correlation between plasmin activity per volume and the amount of CF was seen. Neither did CF protein concentration correlate with the clinical parameters. Periodontal treatment resulted in a dramatic decrease in plasmin activity. In 20 days the activity returned to pre-operative levels in four subjects out of five.     

Gingival crevicular fluid VEGF levels in periodontal health and disease

BACKGROUND: Vascular endothelial growth factor (VEGF), a glycoprotein, has attracted attention as a potential inducer of angiogenesis. It is detectable in periodontal tissues within endothelial cells, plasma cells, and macrophages and in junctional, sulcular, and gingival epithelium. In periodontitis patients, the volume of gingival crevicular fluid (GCF) and the total amount of VEGF collected from diseased sites were greater than from clinically healthy sites. The aim of the present study was to investigate the role of VEGF in periodontal disease progression and to investigate the effect of periodontal therapy on VEGF concentrations in GCF. METHODS: Forty-five subjects were divided into three groups based on gingival index, clinical attachment loss, and radiographic evidence of alveolar bone loss: healthy (group 1), gingivitis (group 2), and chronic periodontitis (group 3). A fourth group consisted of subjects from group 3, 8 weeks after treatment (scaling and root planing). GCF samples collected from each patient were quantified for VEGF levels using enzyme-linked immunosorbent assay. Further, the correlation between VEGF levels in situ and the clinical parameters was analyzed in all groups and was analyzed before and after treatment in the periodontitis group. RESULTS: The highest mean VEGF concentration (99.375 pg/ml) was observed in group 3, and the lowest was observed in group 1 (42.025 pg/ml). Its mean level in group 3 decreased to 54.60 pg/ml after treatment (group 4). Further, GCF VEGF levels showed a positive correlation with all of the clinical parameters. CONCLUSIONS: VEGF levels in GCF increased from health to periodontitis, and periodontal treatment resulted in a reduction in their concentrations. These data indicated that VEGF plays a key role in periodontal disease progression and can be considered a biomarker of periodontal disease progression.     

Gingival crevicular fluid osteopontin levels in periodontal health and disease

BACKGROUND: Osteopontin (OPN), a glycosylated phosphoprotein, is a bone matrix component produced by osteoblasts, osteoclasts, and macrophages as a multifunctional cytokine. OPN anchors osteoclasts to the bone surface, and its absence leads to impaired bone resorption. The aim of the present study was to assess the relation between clinical parameters and concentrations of OPN within gingival crevicular fluid (GCF) from inflamed gingiva and periodontitis sites and, subsequently, after the treatment of periodontitis sites. METHODS: A total of 45 subjects were divided into the following three groups based on modified gingival index (MGI) and Ramfjord periodontal disease index (PDI) scores: healthy (group I), gingivitis (group II), and chronic periodontitis (group III). A fourth group consisted of 15 subjects from group III, 6 to 8 weeks after treatment (i.e., scaling and root planing [SRP]). GCF samples collected from each patient were quantified for OPN using the enzymatic immunometric assay. Further, the correlation between OPN levels in situ with clinical parameters was analyzed in all groups and before and after treatment in periodontitis patients. RESULTS: The highest mean OPN concentration in GCF (14.347 microg/ml) was observed in group III, and the lowest mean OPN concentration in GCF (2.522 microg/ml) was observed in group I. Its levels in group III decreased to 8.419 microg/ml after treatment (group IV). Further, GCF OPN levels in all the groups showed a statistically significant positive correlation with clinical attachment loss (P <0.05). CONCLUSIONS: OPN levels increase in GCF from healthy to periodontitis states, and periodontal treatment results in the reduction of OPN levels. The data indicate that OPN may play a key role in, and could be considered a biomarker of, periodontal disease progression.     

Gingival crevicular fluid fibronectin degradation in periodontal health and disease

Abstract— The molecular forms of fibronectin (FN) in gingival crevicular fluid of five subjects with at least two sites exhibiting clinical signs of inflammation and pockets of at least 4 mm (test group) and five subjects with clinically healthy periodontium (control group) were investigated. Samples were collected with standard filter paper strips. In the test group samples from both diseased and healthy sites were collected. After collection the test group received one episode of periodontal treatment (scaling and root planing). The sampling and clinical recordings were repeated for the diseased sites after about 2 wk. The crevicular fluid FN was analyzed using sodium dodecyl sulphate gel electrophoresis followed by western blotting with polyclonal antibodies against FN. Both intact FN and FN fragments were found in all samples. A larger proportion of FN was in degraded form in the diseased sites than in the healthy or the treated sites. FN was also degraded into smaller peptide fragments in the diseased than in the treated sites. These results suggest that crevicular fluid FN is partially degraded both in periodontal health and disease and that the degree of degradation of FN increases with periodontal inflammation and decreases with periodontal treatment.     

牙周机械治疗对慢性牙周炎患者龈沟液中疱疹病毒的影响

目的 通过纵向观察慢性牙周炎患者(chronic periodontitis,CP)治疗前及治疗后不同时间点龈沟液中两种疱疹病毒检出率的变化,分析疱疹病毒与牙周炎症的关系,为评价牙周炎治疗效果及判断预后提供一个微生物学手段.方法 应用巢式聚合酶链反应(nested PCR,n-PCR)技术,检测13例CP患者牙周机械治疗前、治疗后2周、2个月、4个月时龈沟液中的人巨细胞病毒(human cytomegalovirus,HCMV)、EB病毒1型(Epstein-Barr vires-1,EBV-1),并分析两种病毒检出率的变化.结果 CP患者在牙周机械治疗后2个月和4个月的菌斑指数、探诊深度和出血指数均较治疗前显著降低(P<0.01),但治疗后4个月的菌斑指数、探诊深度和出血指数均较治疗后2个月显著升高(P<0.05).CP患者治疗前HCMV、EBV的检出率分别为42%(33/78)、14%(11/78);EBV阳性位点多伴有HCMV感染,11个EBV(+)的位点中9个位点伴HCMV(+).HCMV、EBV检出的位点多是深牙周袋位点,14个深牙周袋位点中13个HCMV(+);11个EBV(+)位点中9个为深牙周袋位点.CP患者在治疗后2周时HCMV、EBV的检出率降至最低(分别为8%、0);治疗后2周、2个月、4个月HCMV和EBV的检出率均明显低于治疗前(P<0.01),HCMV在治疗后2个月的检出率(15%)较治疗后2周时(8%)回升,但差异无统计学意义(P=0.133).结论 疱疹病毒感染在CP发病中可能有一定作用;CP患者在牙周机械治疗后疱疹病毒的变化早于临床指标的变化,治疗后2个月内即应开始定期复查、复治.     

Gingival crevicular fluid and plasma oxidative stress markers and TGM-2 levels in chronic periodontitis

ObjectiveThis study was aimed to evaluate the gingival crevicular fluid (GCF) and plasma transglutaminase-2 (TGM-2), total antioxidant capacity (TAC), total oxidant status (TOS), ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) in patients with chronic periodontal disease.Materials and methodsTwenty patients with chronic periodontitis (CP), 20 patients with gingivitis and 20 healthy subjects were enrolled in the study. Clinical periodontal parameters including probing depth, clinical attachment level, plaque index and papillary bleeding index were recorded. GCF and plasma levels of TGM-2, TAC, TOS, TBARS and FRAP were analyzed.ResultsGCF TGM-2 was significantly lower in CP group than in gingivitis patients (P = 0.006). GCF FRAP in CP and gingivitis groups was significantly lower than in healthy subjects (P < 0.001). Plasma FRAP level was lower in gingivitis group when compared to healthy subjects (P = 0.003). There was no significant difference in GCF and plasma TAC, TOS, TBARS and plasma TGM-2 levels among the study groups (P > 0.05). GCF TGM-2 level was positively correlated with GCF TAC and negatively correlated with CAL.ConclusionsDecreased FRAP in GCF and plasma indicating lower antioxidant status of CP patients might suggest the role of oxidative stress in periodontitis. GCF TGM-2 data might suggest that TGM2 is associated with stabilization of the extracellular matrix and wound healing in periodontium rather than gingival inflammation.     

SPR牙周基础治疗对慢性牙周炎患者龈沟液C反应蛋白的影响

目的: 探讨SPR牙周基础治疗对慢性牙周炎患者牙周炎症控制、炎症指标龈沟液C反应蛋白的影响。方法: 将80例慢性牙周炎患者平均分为2组,实验组采用SPR牙周基础治疗,对照组仅接受龈上洁治术,比较2组治疗后3个月、6个月牙周指数[包括平均探诊深度（PO）、临床附着水平（CAL）和龈沟出血指数（SBI）]和龈沟液C反应蛋白（CRP）的变化。采用 SPSS 20.0软件包对数据进行统计学处理。结果: 2组患者治疗后3个月、6个月牙周指标SBI、PD 和 CAL以及龈沟液CRP均降低。实验组治疗后3个月、6个月实验组牙周指标及龈沟液CRP显著低于对照组（P<0.01）;对照组治疗后3个月牙周指标及龈沟液CRP与治疗前有显著差异,但治疗后6个月牙周指标、龈沟液CRP逐渐上升。结论: 相对于龈上洁治,完整的SPR牙周基础治疗更能降低慢性牙周炎患者龈沟液C反应蛋白的水平。