Crevicular fluid matrix metalloproteinase‐8, ‐13, and TIMP‐1 levels in type 2 diabetics

Oral Diseases (2010) 16 , 476–481 Objectives: To evaluate whether type 2 diabetes mellitus (DM) enlarged and if so the quantum of such increase in the gingival crevicular fluid (GCF) levels of matrix metalloproteinase‐8 (MMP‐8), MMP‐13 and tissue inhibitor of metalloproteinases‐1 (TIMP‐1). Methods: Subjects (n = 73) were divided into five groups as follows: 12 DM patients with gingivitis (DM‐G), 12 DM patients with periodontitis (DM‐P), 12 systemically healthy patients with gingivitis (H‐G), 13 systemically healthy patients with periodontitis (H‐P) and 24 periodontally, systemically healthy volunteer subjects (H‐C). Full‐mouth clinical periodontal measurements were performed at six sites per tooth. Gingival crevicular fluid samples were obtained from two sites representing the clinical periodontal diagnosis in single‐rooted teeth. Gingival crevicular fluid levels of MMP‐8, MMP‐13 and TIMP‐1 were analysed by immunofluorometric MMP assay (IFMA), enzyme‐linked immunosorbent assay (ELISA). Data were tested statistically by parametric tests. Results: All clinical periodontal measurements were similar in both diabetic and systemically healthy patients with periodontal disease (all P > 0.05). Total amounts of MMP‐8 in GCF samples were significantly lower in H‐C group than DM‐G, DM‐P, H‐P groups (all P < 0.05). Matrix metalloproteinase‐13, TIMP‐1 total amounts were similar in study groups (P > 0.05). Diabetes mellitus patients exhibited similar levels of MMP‐8, MMP‐13, TIMP‐1 with systemically healthy gingivitis/periodontitis patients (P > 0.05). Conclusions: Within the limits of this study, DM does not seem to significantly affect GCF levels of MMP‐8, MMP‐13, TIMP‐1 or clinical periodontal status.     

Expression of vascular endothelial growth factor in human periodontal tissues: comparison of healthy and diabetic patients

BACKGROUND: Vascular endothelial growth factor (VEGF) induces proliferation of endothelial cells, stimulates angiogenesis, and increases vascular permeability, but information about its role in periodontal lesions is limited. The aim of this study is to determine the association between VEGF expression in healthy and periodontally diseased tissues of healthy and diabetic patients. METHODS: Ten systemically healthy and 10 Type 2 diabetic patients (DM) all diagnosed with periodontitis were enrolled into the study. Gingival samples were collected from both periodontal and healthy sites in all patients. Each patient served as his/her own control. Additionally, 10 people without any systemic or periodontal diseases were enrolled as a negative control group. RESULTS: In the negative control group tissue samples, no VEGF expression was observed. Among the 10 systemically healthy people, no evidence of VEGF was observed in healthy gingival samples, but was found in diseased tissues in 2 cases. In the diabetic patients, VEGF was observed in 4 healthy gingival tissues and in 6 periodontal sites. VEGF was intensely present in monocytes and macrophages. CONCLUSION: The results of this study show that VEGF is increased in gingival tissues of diabetic patients, especially those with periodontal disease.     

牙周基础治疗对2型糖尿病伴慢性牙周炎患者龈沟液丝氨酸蛋白酶抑制剂和肿瘤坏死因子-α水平的影响

目的 观察牙周基础治疗对2型糖尿病伴发或不伴发慢性牙周炎患者龈沟液(gingival crevicular flu-id,GCF)丝氨酸蛋白酶抑制剂(vaspin)和肿瘤坏死因子-α(TNF-α)水平的影响.方法 本研究包含60个研究对象,分为4组:15例2型糖尿病伴发慢性牙周炎患者为DM-CP组;15例慢性牙周炎不伴发2型糖尿病患者为CP组;15例牙周健康的2型糖尿病患者为DM组;15例牙周及全身系统均健康的个体为CTRL组.治疗前与牙周基础治疗8周后取样GCF并检测牙周临床指标.通过ELISA法检测GCF样本中vaspin和TNF-α的水平.结果 治疗前慢性牙周炎组GCF中vaspin和TNF-α水平显著高于牙周健康组(P<0.05),治疗后慢性牙周炎组GCF中vaspin和TNF-α水平显著降低(P<0.05).各组vaspin总量与TNF-α总量、糖化血红蛋白水平、牙龈指数以及探诊深度在统计学上存在正相关关系(P<0.05).结论 牙周基础治疗能明显降低慢性牙周炎患者GCF中vaspin和TNF-α的水平.提示GCF中vaspin和TNF-α可作为糖尿病、牙周炎诊断及其预后的炎性标志物.     

The effect of non-surgical periodontal therapy on peroxidase activity in diabetic patients: a case-control pilot study

Objective: To evaluate the effect of periodontal therapy on clinical parameters as well as on total salivary peroxidase (TSP) activity and myeloperoxidase (MPO) activity in the gingival crevicular fluid (GCF) of patients with type 2 diabetes mellitus (DM2) and of systemically healthy individuals.
Material and Methods: Twenty DM2 subjects with inadequate metabolic control (test group) and 20 systemically healthy individuals (control group), both groups with chronic periodontitis, were enrolled. Periodontal clinical parameters, namely periodontal probing depth (PD), clinical attachment level (CAL), visible plaque index (VPI), bleeding on probing (BOP), gingival bleeding index (GBI) and presence of suppuration (SUP), as well as TSP activity and GCF MPO activity, were assessed before and 3 months after non-surgical periodontal therapy.
Results: At baseline and 3 months post-treatment, the test group presented a higher percentage of sites with VPI and BOP ( p <0.01). MPO activity in the GCF presented lower values ( p <0.05) for the test group at both baseline and the post-treatment period. The periodontal treatment resulted in a significant improvement of most clinical and enzymatic parameters for both groups ( p <0.05).
Conclusions: In both groups, the periodontal therapy was effective in improving most clinical parameters and in reducing salivary and GCF enzymatic activity. The diabetic individuals presented lower MPO activity in the GCF.     

Chemerin as a Novel Crevicular Fluid Marker of Patients With Periodontitis and Type 2 Diabetes Mellitus

Background: The objectives of the present study are to: 1) determine whether gingival crevicular fluid (GCF) chemerin is a novel predictive marker for patients with chronic periodontitis (CP) with and without type 2 diabetes mellitus (t2DM); 2) analyze the relationship between chemerin and interleukin (IL)‐6 in periodontally healthy individuals and in patients with CP and with and without t2DM; and 3) evaluate the effect of non‐surgical periodontal therapy on GCF chemerin levels. Methods: Eighty individuals were split into four groups: 20 who were systemically and periodontally healthy (CTRL), 20 with t2DM and periodontally healthy (DM‐CTRL), 20 systemically healthy with CP (CP), and 20 with CP and t2DM (DM‐CP). Individuals with periodontitis were treated with non‐surgical periodontal therapy. GCF sampling procedures and clinical periodontal measures were performed before and 6 weeks after treatment. Enzyme‐linked immunosorbent assay was used to measure chemerin and IL‐6 levels. Results: Greater values for GCF chemerin and IL‐6 levels were found in CP groups than in periodontally healthy groups, in DM‐CP than in CP, and in DM‐CTRL than in CTRL (P <0.008). GCF chemerin and IL‐6 levels decreased following therapy in CP groups (P <0.02). A comprehensive overview of all groups showed a statistically significant positive correlation of chemerin with IL‐6, glycated hemoglobin, sampled‐site clinical attachment level, and gingival index (P <0.05). Conclusions: In this study, periodontitis and t2DM induced aberrant secretion of chemerin, and non‐surgical periodontal therapy influenced the decrease of GCF chemerin levels in patients with CP with and without t2DM. Furthermore, it suggests GCF chemerin levels may be considered a potential proinflammatory marker for diabetes, periodontal disease, and treatment outcomes.     

Collagenases in gingival crevicular fluid in type 1 diabetes mellitus

BACKGROUND: Studies have demonstrated that high levels of collagenase activity in gingival crevicular fluid (GCF) are associated with degradation of periodontal tissues in progressive periodontitis compared to periodontally healthy tissues. Because the activation of collagenases is an important issue in periodontitis, we have studied the activation of collagenase in gingival crevicular fluid samples of diabetic patients. METHODS: Collagenase activity was studied in human gingival crevicular fluids. Twenty-two poorly controlled diabetic patients (e.g., blood glucose: 11.0+/-0.7 mmol/l; hemoglobin A1c [HbA1c]: 9.6%+/-0.3%) and five well-controlled diabetic patients were compared to six chronic periodontitis subjects and five healthy controls. Collagenase activity against type I collagen was measured using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis quantitated by laser densitometry. RESULTS: The poorly controlled diabetic patients had more alveolar bone loss than the well-controlled diabetic subjects and controls (P<0.001; t test). The activity of collagenases in GCF in poorly controlled diabetic patients was similar to that seen in chronic periodontitis subjects (P>0.05) but higher than in healthy controls (P<0.01; t test), whereas there was no difference between the well-controlled diabetic subjects and systemically healthy controls (P>0.05; t test). CONCLUSION: Poorly controlled diabetes is strongly related to periodontal tissue destruction, and collagenases in GCF may mediate and reflect this effect.     

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.     

Vascular endothelial growth factor (VEGF) levels of gingiva and gingival crevicular fluid in diabetic and systemically healthy periodontitis patients

It has been demonstrated that diabetes mellitus (DM) may have an inductive effect on the vascular endothelial growth factor (VEGF) levels of periodontium during periodontal disease. The aim of this study is to confirm this phenomenon, investigating whether it is also valid for diabetic periodontitis patients under good metabolic control. Sixteen type II DM patients, all with a glycosylated hemoglobin (HbA1c) value less than 7 (test), and 15 systemically healthy (control) chronic periodontitis patients were included in the study. The VEGF concentrations in the gingival supernatants and gingival crevicular fluid (GCF) samples of the study groups were measured by enzyme-linked immunosorbent assay. The data were analyzed by Student’s t test in statistical means. The VEGF levels were significantly higher in the gingival supernatants of the test group (55.89 ± 8.11 pg/ml) than that of the control group (24.81 ± 2.04 pg/ml; p < 0.01). However, there was no statistically significant difference in the VEGF levels of GCF between the study groups (38.96 ± 4.89 pg/ml in the test and 32.20 ± 4.02 pg/ml in the control group; p > 0.05). Our study confirms that DM affects the VEGF levels of periodontal soft tissues in periodontal disease, and our results also suggest that this effect may not be influenced by the metabolic control of DM.     

Evaluation of t-PA, PAI-2, IL-1beta and PGE(2) in gingival crevicular fluid of rheumatoid arthritis patients with periodontal disease

AIMS: This study was undertaken to compare periodontal conditions, gingival crevicular fluid (GCF) levels of tissue-type plasminogen activator (t-PA), its inhibitor plasminogen activator inhibitor-2 (PAI-2), interleukin-1beta (IL-1beta), prostaglandin E(2) (PGE(2)) in rheumatoid arthritis (RA) patients and control groups. METHODS: Twenty-three RA patients, 17 systemically healthy patients with periodontal disease (PD), and 17 systemically and periodontally healthy subjects were recruited. GCF samples were obtained from two single-rooted teeth. Full-mouth clinical periodontal measurements were recorded at six sites/tooth. GCF samples were analysed using relevant ELISA kits. Data were tested statistically by appropriate tests. RESULTS: Total amounts of t-PA, PAI-2 and PGE(2) in GCF samples of the healthy control group were significantly lower than the other groups (p<0.05). The RA group exhibited a higher total amount of t-PA in GCF samples than the PD group (p<0.05). PAI-2, IL-1beta and PGE(2) total amounts were similar in RA and PD groups (p>0.05). CONCLUSION: The coexistence of RA and periodontitis does not seem to affect clinical periodontal findings or systemic markers of RA. Similar inflammatory mediator levels in RA and PD groups, despite the long-term usage of corticosteroids, non-steroidal anti-inflammatory drugs, suggest that RA patients may have a propensity to overproduce these inflammatory mediators.     

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.     

Superoxide dismutase activity in gingiva in type-2 diabetes mellitus patients with chronic periodontitis

OBJECTIVE: Antioxidant defence reduces in diabetes mellitus (DM) and periodontitis. This study investigates antioxidant enzyme; superoxide dismutase (SOD) activity in gingiva and blood glucose and lipid levels in type-2 DM patients and systemically healthy individuals with chronic periodontitis (CP). MATERIALS AND METHODS: Periodontal parameters, blood glycated-haemoglobin (HbA1c), glucose and lipid levels, and gingival-SOD activities (spectrophotometric assay) were measured in 17 DM patients with CP (DMCP), 17 systemically healthy CP patients, 18 periodontally healthy DM patients (DMPH), and 17 healthy controls (PH). RESULTS: Periodontal parameters were higher in periodontitis groups than the controls (p<0.05), while there was no difference between the periodontitis groups and between the control groups. HbA1c, glucose, and triglyceride levels were higher in diabetic groups than the non-diabetic groups (p<0.05). Low-density lipoprotein (LDL), very-LDL and cholesterol values of the DMCP group did not significantly differ from the CP group. No differences existed between diabetic patients with and without periodontitis in HbA1c, glucose, and lipid levels and the same was true for non-diabetic patients with and without periodontitis. Gingival-SOD activity was lower in periodontitis groups than the matched control groups (p<0.05). DMPH group had the highest and CP group had the lowest SOD levels. There were correlations between periodontal parameters, gingival-SOD activity, HbA1c, glucose and high-density lipoprotein (HDL) levels. CONCLUSION: The results suggest that gingival-SOD activity increases in diabetes and decreases in periodontitis and relations may exist between gingival-SOD activity, periodontal status, HbA1c, glucose and HDL levels. The higher gingival-SOD activity in diabetes may be attributed to an adaptive mechanism in the tissue.     

Heightened gingival inflammation and attachment loss in type 2 diabetics with hyperlipidemia

BACKGROUND: Our previous studies in diabetic (DB) rats suggest that hyperlipidemia may cause a dysregulation of the cellular and local cytokine response to periodontitis (AP). The objective of the present study was to determine if diabetes has a similar dysregulatory effect on the gingival response to AP in humans. METHODS: Peripheral blood, as well as gingival tissue (GT) and gingival crevicular fluid (GCF), was obtained from a total of 35 patients who were categorized into the following groups based on level of diabetic (type 2) control and presence or absence of adult periodontitis (AP): group 1, systemically and periodontally healthy (n = 6); group 2, systemically healthy with adult periodontitis (n = 7); group 3, well-controlled diabetes and periodontally healthy (n = 6); group 4, well-controlled diabetes with adult periodontitis (n = 5); group 5, poorly controlled diabetes and periodontally healthy (n = 5); group 6, poorly controlled diabetes and adult periodontitis (n = 6). All subjects were given a thorough periodontal examination, including probing depths (PD), clinical attachment levels (CAL), gingival index (GI), plaque index (PI), and vertical bitewing radiographs. Blood studies included levels of glycated hemoglobin (HbA1c), triglycerides (TG), cholesterol (CHL), low-density lipoproteins (LDL), and high-density lipoproteins (HDL). The levels of interleukin-1 beta (IL-1beta) in GCF and GT, interleukin-6 (IL-6), and platelet-derived growth factor AB (PDGF-AB) in GT from patients in each experimental group were analyzed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Our results indicate that all clinical indices except PI were significantly elevated in the poorly controlled and well-controlled diabetics, compared to systemically healthy patients, but only in the subjects without preexisiting AP (Tukey's multiple comparisons, P <0.05). Pairwise linear regression analysis revealed significant (P <0.01) positive associations between periodontal inflammation (PD, CAL, PI, GI) and levels of GCF IL-1beta, GT IL- 1beta GT IL-6, but not GT PDGF; moreover, GT IL-6 levels were significantly associated (P<0.05) with GT IL-1beta. As TG levels increased in the non-AP patients (group 1 < group 3 < group 5), there was a trend, not significant, for increased GCF IL-1beta levels and increased gingival inflammation. Interestingly, periodontitis resulted in increased PDGF-AB levels in the gingiva of systemically healthy and well-controlled diabetes patients, but this increase was obtunded in poorly controlled diabetes patients. CONCLUSIONS: This confirms our earlier work in the diabetic rat model. These studies indicate that decreased metabolic control in type 2 diabetics results in increased serum triglycerides and has a negative influence on all clinical measures of periodontal health, particularly in patients without preexisting periodontitis. Levels of the cytokine IL- 1beta showed a trend for increasing as diabetic control diminished. In contrast, levels of the growth factor PDGF, which normally increase in periodontitis, decreased in poorly controlled diabetics with periodontitis. These studies suggest a possible dysregulation of the normal cytokine/growth factor signaling axis in poorly controlled type 2 diabetics that may contribute to periodontal breakdown/diminished repair.     

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.     

Periodontal status and cytoplasmic enzyme activities in gingival crevicular fluid of type 2 diabetic and/or obese patients with chronic periodontitis

Type 2 diabetes mellitus and obesity are the most common nutritional disorders in developed and developing countries. Increased prevalence of periodontal disease is a well-known complication of type 2 diabetes mellitus (DM). As obesity is generally the first step toward type 2 diabetes mellitus, it is possible to find exacerbated periodontal disease in obese patients, also. The purpose of this cross-sectional study was to investigate the periodontal status and aspartate aminotransferase and lactate dehydrogenase enzyme activities in gingival crevicular fluid (GCF) of type 2 diabetic and/or obese chronic periodontitis patients. A total of 39 chronic periodontitis patients participated in the study. The study population was divided into four groups according to body mass index and type 2 DM status: 1) type 2 DM obese patients, n = 8; 2) type 2 DM patients, n = 12; 3) obese patients, n = 8; 4) systemically healthy control group, n = 11. Enzyme activities in gingival crevicular fluid and periodontal status were evaluated. No significant differences in age, gingival index, plaque index, aspartate aminotransferase and lactate dehydrogenase enzyme activities were observed, but probing depths were significantly higher in the DM groups than in the control group. Obesity did not seem to be a significant factor in any parameters evaluated. The present study showed increased probing depth values for the diabetic groups but failed to show any significant relation between obesity and enzyme activity or periodontal status. However, the slightly increased probing depth values in the obese groups might be a clue to an impaired immune response and predisposition to periodontitis in that patient group.     

Diabetes‐Associated Periodontitis Molecular Features in Infrared Spectra of Gingival Crevicular Fluid

Background: It has been established previously that infrared spectroscopy (IRS) can be used to identify periodontitis‐specific molecular signatures in gingival crevicular fluid (GCF) and to confirm clinical diagnoses. This follow‐up study is designed to assess whether this novel technique is also able to differentiate diseased from healthy sites in patients with diabetes mellitus (DM) by analyzing the molecular fingerprints embedded in the GCF. Methods: A total of 65 patients with DM with moderate‐to‐severe chronic periodontitis (CP) was recruited, and 15 individuals without DM (65 sites) without periodontal diseases were used as control. Clinical examination and GCF samples were taken from a total of 351 sites, including periodontitis (109), gingivitis (115), and healthy (127) sites. Corresponding absorption spectra of GCF samples were acquired and processed, and the relative contributions of key functional groups in the infrared spectra were identified and analyzed. The qualitative assessment of clinical relevance of these GCF spectra was interpreted with multivariate statistical analysis: linear discriminant analysis (LDA). Results: Spectral analysis revealed several molecular signatures representing vibrations in protein (amide I and II), lipid ester, and sugar moieties in the GCF of patients with DM with CP and non‐DM controls. The diagnostic accuracy for distinction between healthy and CP sites in patients with DM determined by LDA of GCF spectra was 95.3% for the training set of samples and 87.5% for the validation set. Additional LDA of GCF spectra from healthy sites of non‐DM controls and patients with DM revealed 100% diagnostic accuracy for the training set and 86.7% for the validation set. The regions robotically selected by LDA for the two analyses were slightly different in that first LDA identified major regions clustered with the side chain vibrations originating from protein and DNA contents, whereas the second was predominantly the glycation and protein components. Conclusion: IRS is a feasible method to differentiate disease‐specific molecular signatures in GCF in the presence of DM and to generate a complex biochemical profile of GCF to identify DM‐specific spectral features.     

Fluid dynamics of gingiva in diabetic and systemically healthy periodontitis patients

OBJECTIVES: The influence of diabetes mellitus (DM) on the fluid dynamics of periodontium has not been reported in periodontal disease. The objectives of this study were (i) to investigate the alterations in the fluid dynamics of periodontium in diabetic periodontitis patients, and present the association of this phenomenon with the metabolic control of DM; (ii) to reveal any correlation between the fluid dynamics of periodontium and clinical signs of periodontal disease in DM and periodontitis. DESIGN: Fifteen well-controlled diabetic chronic periodontitis patients (Group 1), 14 systemically healthy chronic periodontitis patients (Group 2), and 14 systemically and periodontally healthy individuals were included in the study. Gingival crevicular fluid volume (GCF-V) and gingival tissue osmotic pressure (GOP) were used as the parameters of periodontal fluid dynamics. GCF-V was measured by a Periotron device, while GOP was measured by a digital osmometer. Silness-L?e plaque index (PI), L?e-Silness gingival index (GI) and clinical attachment loss (AL) levels were recorded to determine the periodontal health status. RESULTS: PI, GI and AL were higher in Groups 1 and 2 than in Group 3 (P<0.05), but similar between Groups 1 and 2 (P>0.05). Increased GCF-V and GOP were observed in Groups 1 and 2 compared with Group 3 (P<0.01), and the increase in Group 1 was greater than that in Group 2 (P<0.01). There were strong positive correlations between GCF-V and GOP in all three groups: between GI and GCF-V and GI and GOP in Groups 1 and 2; and between AL and GCF-V and AL and GOP in Groups 2 and 3. CONCLUSION: The results suggest that (i) DM may have an additive influence on the fluid dynamics of periodontium in the presence of periodontal disease; (ii) this phenomenon may not be prevented by the metabolic control of DM; (iii) the clinical signs of periodontal disease may be affected by the fluid dynamics of periodontium in both DM and periodontitis.     

Glycated Albumin and Calprotectin Levels in Gingival Crevicular Fluid From Patients With Periodontitis and Type 2 Diabetes

Background: Patients with diabetes mellitus (DM) have a high prevalence of periodontitis. Periodontitis in these patients is characterized by severe inflammation and tissue breakdown, and its diagnosis is important for cures of periodontitis and DM. The purpose of this study is to investigate the levels of glycated albumin (GA), a DM marker, and calprotectin, an inflammatory marker, in gingival crevicular fluid (GCF) from patients with periodontitis and DM (DM‐P). Methods: The 78 participants in this study were patients with DM, chronic periodontitis (CP), DM‐P, and healthy individuals (H). GCF and blood were collected, and GA and calprotectin in GCF were analyzed using Western blotting and enzyme‐linked immunosorbent assay. Levels were compared among H, DM, CP, and DM‐P groups. Blood GA and glycated hemoglobin (HbA1c) were measured, and the correlation among GCF GA and blood HbA1c or GA levels was investigated. Receiver operating characteristic (ROC) analysis for GCF GA to predict DM was performed. Results: GA was identified in GCF, and its amount/concentration in GCF samples from DM and DM‐P were significantly higher than those of non‐DM groups (H and CP). Calprotectin amounts in GCF from CP and DM‐P were significantly higher than in H and DM groups. GCF GA level was positively correlated with blood HbA1c and GA level. ROC analysis of GCF GA showed an optimal cutoff value to predict DM. Conclusions: GA showed a high level in GCF from patients with DM. Examination of GA and calprotectin in GCF may be useful for predicting DM‐P.     

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.     

Levels of interleukin-1 beta, -8, and -10 and RANTES in gingival crevicular fluid and cell populations in adult periodontitis patients and the effect of periodontal treatment

BACKGROUND: Various cytokines have been identified at sites of chronic inflammation such as periodontitis. Cytokines are synthesized in response to bacteria and their products, inducing and maintaining an inflammatory response in the periodontium. The purpose of the present study was to investigate the involvement of interleukin-1 beta (IL-1 beta), IL-8, and IL-10 and RANTES (regulated on activation, normally T cell expressed and secreted) and the cell populations associated with the immune response in destructive periodontitis, as well as the effect of periodontal therapy on cytokine levels in gingival crevicular fluid (GCF). METHODS: Data were obtained from 12 patients with moderate to advanced periodontitis and 6 healthy controls. Patients presenting at least 2 sites with > or =2 mm clinical attachment loss were included in the destructive periodontitis group. After monitoring for 4 months, only 6 patients showed destructive periodontitis and GCF samples and soft tissues biopsies were collected from these patients. GCF samples and biopsies were collected both from active (12 CGF samples and 6 biopsies) and inactive (12 CGF samples and 6 biopsies) sites. The comparison with healthy controls was carried out by collecting GCF samples from 6 healthy volunteers (12 samples) and biopsies during the surgical removal of wisdom teeth. In periodontal patients, clinical data and GCF samples were obtained prior to periodontal treatment (72 samples) and 2 months after periodontal therapy (72 samples). GCF was collected using a paper strip; eluted and enzyme-linked immunoabsorbent assays (ELISA) were performed to determine cytokine levels. The inflammatory infiltrate was analyzed by immunohistochemistry of gingival biopsy samples with monoclonal antibodies against CD3, CD8, CD4, CD11c, and CD19 antigens. RESULTS: Cellular components of the inflammatory infiltrate include B and T lymphocytes and monocyte/macrophages. Active sites contained a higher number of B lymphocytes and macrophages. IL-8 and IL-1 beta and RANTES in GCF were detected in the majority of sites from periodontal patients (100%, 94% and 87%, respectively); IL-10 was found in only 43%. IL-8 was the only cytokine detected in the GCF (75%) of the control group. Moreover, IL-1 beta levels were significantly higher in active sites versus inactive sites (P <0.05). IL-8 and IL-10 and RANTES were increased in active sites; however, differences were not significant (P>0.05). A positive correlation between the IL-8 and RANTES (r = 0.677, P<0.05) was observed in periodontitis patients. Periodontal therapy reduced the total amount of IL-1 beta, IL-8, and IL-10 and RANTES. Data showed a weak correlation between the clinical parameters and the total amount of cytokines in periodontitis. CONCLUSIONS: These data suggest that the amount of crevicular IL-1 beta, IL-8, and IL-10 and RANTES is associated with periodontal status. Removal of the bacterial plaque reduces the antigenic stimuli and consequently could modulate the chemokines present in GCF. We propose that the dynamic interactions between cytokines, their production rates, and their quantity could represent factors controlling the induction, perpetuation, and collapse of the cytokine network present in the periodontal disease.     

Plasminogen activator system in smokers and non-smokers with and without periodontal disease

BACKGROUND: The present study assessed levels of plasminogen activator (PA) system proteins in gingival crevicular fluid (GCF) and serum of chronic gingivitis, chronic periodontitis patients and periodontally healthy subjects and evaluated how smoking influenced these levels. METHODS: Twenty chronic gingivitis; 20 chronic periodontitis patients and 20 periodontally healthy volunteers were consecutively recruited according to the inclusion criteria so that exactly half of the subjects in each category were smokers. GCF samples from four sites together with serum samples were obtained from each subject. GCF levels of tissue type PA (t-PA), urokinase type PA (u-PA), PA inhibitor-1 (PAI-1) and PA inhibitor-2 (PAI-2) and serum concentrations of cotinine, u-PA and PAI-1 were analysed by enzyme-linked immunosorbent assay. RESULTS: The only statistically significant difference between smokers and non-smokers was a lower GCF PAI-2 concentrations in healthy smokers compared with healthy non-smokers (p<0.01). Gingivitis and periodontitis patients had higher GCF concentrations of PAI-2 than healthy subjects (p<0.002 and p<0.02 respectively). The ratio of u-PA:PAI-1 and t-PA:PAI-1 were significantly higher in GCF of smokers with periodontitis compared with "healthy" smokers, whereas the ratio of t-PA:PAI-2 was significantly lower in smokers with periodontal disease (p<0.05). CONCLUSIONS: GCF levels of the PA system proteins are increased in chronic gingivitis and periodontitis compared with healthy gingiva. Smoking had only subtle effects on the GCF PA system proteins with the exception of PAI-2, and the balance of activators and inhibitors. These findings suggest one mechanism whereby smoking may exert detrimental effects on the periodontal tissues.