Vascular endothelial growth factor in gingival tissues and crevicular fluids of diabetic and healthy periodontal patients

BACKGROUND: Periodontal disease is one of the major oral problems encountered in patients with diabetes mellitus (DM). Vascular changes, neutrophil dysfunction, altered collagen synthesis, and genetic predisposition observed in DM may contribute to periodontitis; and the vascular alterations observed in such patients may depend on vascular endothelial growth factor (VEGF) actions. Few reports are available about the mechanism of neovascularization and the angiogenic factors that contribute to the periodontal pathology and the role of VEGF in periodontal diseases. The aim of this study is to compare VEGF expression in healthy and periodontally diseased tissues with gingival crevice fluid (GCF) of healthy persons and diabetic patients. METHODS: Gingival tissue and GCF samples were collected from sites of periodontitis in 10 healthy subjects and in 10 type 2 diabetic patients, and from the sites of healthy gingiva within the same groups. Therefore, each patient became his/her own control. Additionally, 10 people without any systemic or periodontal diseases were enrolled, forming a negative control group. Thus, a total of 50 tissue and 50 GCF samples were provided. RESULTS: No VEGF staining was observed in the negative control group or in the systemically healthy people's healthy tissue samples, whereas four samples of diabetic patients showed positive staining (P < 0.05). However, VEGF was revealed in two tissue samples of periodontal sites of systemically healthy people and in six samples of the diabetic patients (P > 0.05). In all test groups, GCF VEGF levels were higher in periodontal sites (P < 0.05) than in healthy sites. CONCLUSION: The results of this study showed that VEGF is increased in all periodontal tissues of both groups and in the healthy sites of diabetic patients. Additionally, GCF VEGF values increased in periodontal sites of all test groups.     

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.     

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 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.     

Increased Bacterial Invasion and Differential Expression of Tight‐Junction Proteins,Growth Factors,and Growth Factor Receptors in Periodontal Lesions

Background: Many pathogens are known to modulate epithelial physical barriers, particularly tight‐junction (TJ) proteins, to enter host cells and/or tissues. Growth factors have been implicated in the regulation of TJ proteins. The aim of this study is to determine differences in the levels of TJ proteins, growth factors, and their receptors in relation to bacterial invasion in diseased gingival tissues obtained from patients with periodontitis. Methods: The presence of bacteria and expression of junctional adhesion molecule (JAM)‐A, occludin, epidermal growth factor (EGF), keratinocyte growth factor (KGF), insulin‐like growth factor‐I (IGF‐I), EGF receptor, KGF receptor, and IGF‐1 receptor (IGF‐1R) were evaluated in gingival tissues from healthy (n = 10) and diseased (n = 10) sites in patients with periodontitis by in situ hybridization and immunohistochemistry. Results: The bacterial invasion of gingival tissue was increased in periodontal lesions compared with healthy sites. Although the levels of JAM‐A and occludin were not significantly different between the healthy and diseased sites, aberrant cytoplasmic expression of JAM‐A and occluding was often observed in the lesions. In addition, more leukocytes expressing JAM‐A or occludin were observed within the disease‐associated epithelia. Compared with the healthy sites, the differential expression of KGF, IGF‐I, and IGF‐1R was observed in the periodontal lesions. The levels of TJ proteins showed positive correlations with those of growth factors. Conclusion: The aberrant expression of growth factors and TJ proteins may contribute to increased bacterial invasion and disease progression in periodontal lesions.     

Tumor necrosis factor-alpha expression and detection of apoptosis at the site of chronic periodontitis in AIDS patients

BACKGROUND: Apoptosis, or programmed cell death, is associated with the regulation of the life cell cycle of leukocytes in healthy and diseased states. OBJECTIVES: In the present study, we investigated the presence of apoptosis of mononuclear inflammatory cells in the periodontal lesion from adult periodontitis in healthy control patients and AIDS patients. MATERIALS AND METHODS: Tissue samples adjacent to a 5-6 mm gingival sulcus, measured with a periodontal probe, were obtained during routine periodontal surgical procedures. The direct immuno-peroxidase of digoxigenin-labeled genomic DNA method was used for in situ detection of apoptosis in gingival tissues. RESULTS: Many tumor necrosis factor-alpha (TNFalpha)-positive cells, detected by immunohistochemistry method, were observed in gingival samples of both groups of patients. In addition, a significant lower number ( p < 0.05) of mononuclear apoptotic cells were observed in AIDS patients when compared with healthy control patients. CONCLUSION: These data suggested an important role of the apoptosis of mononuclear cells in the pathogenesis of chronic adult periodontitis in AIDS patients.     

Overexpression of interleukin-1beta and interleukin-6 may play an important role in periodontal breakdown in type 2 diabetic patients

BACKGROUND AND OBJECTIVE: This study evaluated whether the biochemical changes associated with type 2 diabetes modulate the expression of interleukin-1beta, interleukin-6, interleukin-8, and interferon-gamma in sites with chronic periodontitis. MATERIAL AND METHODS: Biopsies were harvested and divided into three groups: group 1, systemically and periodontally healthy subjects (n = 10); group 2, systemically healthy subjects with moderate-to-severe chronic periodontitis (probing depth > 6 mm) (n = 20); and group 3, type 2 diabetic subjects with periodontitis (n = 20). Cytokine levels were assessed in the gingival tissues by enzyme-linked immunosorbent assay analysis. RESULTS: Data analysis demonstrated that the interleukin-1beta, interleukin-6, interleukin-8, and interferon-gamma levels were higher in the presence of periodontal inflammation than in the absence of inflammation, regardless of systemic status. The interleukin-1beta and interleukin-6 levels were higher in diabetic subjects (group 3) than in systemically healthy patients with comparable types of periodontitis (group 2). No difference was observed for the interleukin-8 and interferon-gamma levels between groups 2 and 3. CONCLUSION: Within the limits of this study, it was concluded that type 2 diabetes was associated with increased expression of interleukin-1beta and interleukin-6 in periodontally inflamed tissues of diabetic patients, relative to nondiabetic subjects, and that such overexpression may be involved in the mechanisms by which type 2 diabetes enhances periodontal destruction.     

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.     

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.     

Purine nucleoside phosphorylase activity and expression are upregulated in sites affected by periodontal disease

Batista EL Jr, Deves C, Ayub L, da Silva RG, Filho LCC, Basso LA, Santos DS. Purine nucleoside phosphorylase activity and expression are upregulated in sites affected by periodontal disease. J Periodont Res 2010; 45: 664–671. © 2010 John Wiley & Sons A/S Background and Objective: Purine nucleoside phosphorylase (PNP) is an enzyme that catalyzes the reversible phosphorolysis of purine nucleosides, playing a key role in the purine salvage pathway. Activated T cells seem to rely heavily on PNP to remain functionally active and are particularly sensitive to PNP deficiency. The role of PNP in periodontal tissues has not been characterized thus far. The aim of this study therefore was to assess the activity and expression of PNP in the gingival tissues of periodontitis patients. Material and Methods: Ten patients consecutively admitted for treatment had their periodontal clinical variables recorded and their gingival crevicular fluid collected. After periodontal treatment the patients were seen once a month for plaque and bleeding control, and had their periodontal variables recorded and gingival crevicular fluid collected at 90 and 180 d. Purine nucleoside phosphorylase‐specific activity was assessed using a spectrophotometer through the addition of the PNP substrate analog 2‐amino‐6mercapto‐7‐methyl purine riboside to the gingival crevicular fluid. In parallel, PNP expression was assessed by immunohistochemistry and real‐time PCR in gingival biopsies and cell culture. Results: Purine nucleoside phosphorylase activity was higher in the gingival crevicular fluid of periodontally diseased sites, which was positively correlated with improvements of the clinical variables. Treatment of periodontal disease induced a striking decrease of PNP activity in periodontally diseased sites. Expression of PNP was more pronounced in mononuclear cells and endothelial cells of the gingiva, and the mRNA levels were 5.7‐fold higher in inflamed tissues compared with control samples. Conclusion: Purine nucleoside phosphorylase activity and expression are upregulated in periodontally diseased sites and can be detected in the gingival crevicular fluid.     

A multicenter clinical trial of PerioGardTM in distinguishing between diseased and healthy periodontal sites

Abstract We designed and performed a multicenter clinical trial to determine the relationship between measurements of the level of the enzyme aspartate amino-transferase (AST) in gingival crevicular fluid (GCF) to other measures used to detect periodontal disease and monitor outcome of treatment, including pocket depth and gingival inflammation. 32 periodontitis patients were enrolled at the University of Washington, Seattle, 30 at the University of Florida, Gainesville, and 34 at the University of Illinois, Chicago. 10 periodontally normal control subjects were enrolled at each location. 8 diseased and 4 healthy sites were designated for study in each patient and 8 healthy sites designated in each control subject. Measures of disease included pocket depth, severity of gingival inflammation, and GCF volume. AST levels were measured using the PerioGard^TM test kit. Clinical measurements were made and GCF samples harvested and tested 2X before and 2X after therapy consisting of scaling and root planing under local anesthetic. Specific design and other issues are discussed, including selection of patients and control subjects, sample size, selection of experimental test sites, methods for assessment of diseased and therapeutic improvement, harvesting of GCF, and selection of appropriate biostatistical methods for data analysis. Demographics of the patient populations at the 3 locations are reported. As expected, therapy induced only negligible changes in the measures of disease at healthy sites in control subjects, and relatively minor improvement in healthy sites in patients. In contrast, statistically significant improvement relative to pre-treatment baseline status in all 3 measures of disease was observed for diseased sites at all 3 study locations with all p-values less than 0.0002. The magnitude of improvement was comparable to that reported previously by others. The % of PerioGard-positive sites decreased significantly between the screening baseline and both post-treatment visits for patients at all 3 locations, with p values of 0.0001 to < 0.0008.     

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.     

Ⅱ型糖尿病合并牙周病患者龈沟液中细胞因子/趋化因子的表达水平

目的 探讨Ⅱ型糖尿病合并牙周病患者与单纯牙周病患者龈沟液（gingival crevicular fluid,GCF）中细胞因子/趋化因子的表达水平。 方法 选取伴Ⅱ型糖尿病的牙周病患者52例,单纯牙周病患者40例,用Luminex FLEXMAP3D仪和Human Cytokine/Chemokine试剂盒检测GCF中14种细胞因子/趋化因子的表达水平。 结果 牙周病部位：嗜酸性粒细胞趋化因子、巨噬细胞炎症蛋白-1α、粒细胞-巨噬细胞集落刺激、白介素-6、肿瘤坏死因子-α和白介素-12的浓度,糖尿病组受试者高于非糖尿病组受试者（P<0.0035）。 结论 糖尿病可影响牙周病部位细胞因子/趋化因子的表达,糖尿病可能是牙周病的促进因素。     

The in vivo expression of membrane-bound CD14 in periodontal health and disease

BACKGROUND: Membrane-bound CD14 (mCD14) is a myeloid differentiation antigen expressed on monocytes/macrophages and neutrophils. It is a key molecule responsible for the innate recognition of bacteria by host cells and functions as an important receptor for bacterial lipopolysaccharide. This study investigated the in vivo expression profile and levels of mCD14 in healthy and diseased gingival tissues. METHODS: Gingival biopsies were obtained from 24 patients with chronic periodontitis, including 22 periodontal pocket tissues, 13 clinically healthy tissues, and 18 inflamed connective tissues (i.e., granulation tissues). Gingival biopsies from seven periodontally healthy subjects were used as controls. mCD14 was detected by immunohistochemistry. RESULTS: mCD14 was detected in 21 of 22 periodontal pocket tissues and all other categories of tissues. The mCD14-positive cells were mainly confined to the gingival epithelium-connective tissue interface. The expression levels in periodontally healthy subjects were significantly higher than in the patients. Within the patients, clinically healthy tissues showed greater levels of mCD14 than periodontal pocket tissues and granulation tissues. CONCLUSIONS: mCD14 was commonly expressed in both healthy and diseased gingival tissues and was predominantly confined to the epithelium-connective tissue interface. The positive relationship observed between mCD14 expression levels and periodontal health may imply that mCD14 is associated with favorable host responses to bacterial challenge and contributes to maintaining periodontal homeostasis.     

Vascular endothelial growth factors and progression of periodontal diseases.

BACKGROUND: Tissues become hemorrhagic and edematous coincident to periodontal diseases; however, there is little information concerning the biologic mechanisms which may produce these changes. Vascular endothelial growth factor (VEGF) is a macromolecule which enhances blood vessel growth and permeability. However, there is no information concerning gingival VEGF concentrations within normal or diseased gingiva. The purpose of this study was to assess changes in gingival concentrations of VEGF during initiation and progression of periodontal diseases and compare them to changes in the number of blood vessel profiles and concentration of recognized markers of periodontal disease severity (interleukin-6[IL-6]). METHODS: Normal (non-hemorrhagic gingiva adjacent to a < or =3 mm gingival sulcus) and inflamed gingiva (hemorrhagic gingiva adjacent to a < or =3 mm, 4 to 6 mm, or >6 mm periodontal pocket) were studied. VEGF and IL-6 concentrations were assessed by ELISA and the number of blood vessels determined by histomorphometric techniques. Data were placed into one of the following groups: < or =3 mm, normal; < or =3 mm, diseased; 4 to 6 mm, diseased; and >6 mm, diseased. These groups were compared by factorial ANOVA and Scheffe comparisons. In addition, groups were compared by simple and multiple regression and regression ANOVA to determine possible correlations between them. RESULTS: VEGF and IL-6 concentrations were significantly lower within normal than within diseased gingiva. The number of blood vessel profiles and mean IL-6 concentrations were highest in diseased tissues adjacent to >6 mm sulci and were significantly correlated with sulcular depth (P <0.001). In contrast, VEGF concentrations were highest within diseased gingiva adjacent to 4 to 6 mm periodontal pockets (P <0.001) and were not correlated with sulcular depth. CONCLUSIONS: VEGF may be a factor in initiation and progression of gingivitis to periodontitis, possibly by promoting expansion of the vascular network coincident to progression of the inflammation.     

Detection of Tannerella forsythensis (Bacteroides forsythus) and porphyromonas gingivalis by polymerase chain reaction in subjects with different periodontal status

BACKGROUND: The aim of this study was to determine the prevalence of Tannerella forsythensis (formerly Bacteroides forsythus) and Porphyromonas gingivalis in subgingival plaque samples by using polymerase chain reaction (PCR), and to assess the relationship of these bacteria with different categories of periodontal disease and health. METHODS: Subjects were distributed into 3 groups according to their periodontal diagnosis: group 1, periodontally healthy (N = 10); group 2, periodontitis with probing depth < or = 5 mm (N = 10); group 3, periodontitis with probing depth > 5 mm (N = 10). The subjects in groups 2 and 3 had healthy and diseased periodontal sites. Subgingival plaque samples were obtained using paper points inserted into periodontal pockets (diseased sites) and into healthy gingival sulci (healthy sites) of the same subject. RESULTS: The distribution of bacteria differed in healthy and diseased sites. T. forsythensis (B. forsythus) was not detected in any sample from healthy sites in any group but was detected in 70% and 100% of diseased sites in groups 2 and 3, respectively. P. gingivalis was detected in only one sample from a healthy site (group 2), and in the diseased sites, its prevalence was 40% (group 2) and 90% (group 3). In addition, T. forsythensis (B. forsythus) and P. gingivalis were both detected in 30% and 90% of the diseased sites in groups 2 and 3, respectively. CONCLUSION: These results indicate a possible association between periodontal disease and the presence of T. forsythensis (B. forsythus) and/or P. gingivalis.     

Correlations between pentraxin 3 or cytokine levels in gingival crevicular fluid and clinical parameters of chronic periodontitis

The gingival crevicular fluid (GCF) contains various biomarkers, such as interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α (TNF-α), and IL-10, among others. These cytokines have been reported to correlate with gingival inflammation and periodontal status. Therefore, the analysis of GCF may be useful for the diagnosis of periodontal status. Pentraxin 3 (PTX3) is the first identified long pentraxin, and is released by several cell types in response to proinflammatory signals. The aim of this study was to determine the levels of IL-1β, IL-6, IL-8, TNF-α, IL-10 and PTX3 in GCF from diseased and healthy sites in patients with chronic periodontitis. Cross-sectional clinical data were obtained from 50 patients with chronic periodontitis. GCF samples were collected with paper strips from one periodontal diseased site and one periodontally healthy site per subject. The levels of IL-1β, IL-6, IL-8, IL-10 and TNF-α were determined using a multiplexed bead immunoassay, and the PTX3 level was measured using an enzyme-linked immunosorbent assay. Mean clinical parameters were significantly higher at diseased sites (P < 0.01) as compared to healthy sites, and the mean levels of PTX3, IL-1β, IL-6, IL-8, IL-10 and TNF-α were higher in diseased sites (P < 0.01) than in healthy sites. There were strong correlations between PTX3 or IL-1β and periodontal status. These results suggest that GCF PTX3 levels might be useful as a diagnostic marker for periodontal disease.     

糖尿病患者牙髓组织病变及IL-8、VEGF表达的研究

目的研究糖尿病患者(diabetes mellitus,DM)和正常人牙髓血管形态结构及IL-8含量、VEGF表达的差异。方法DM组30例为Ⅱ型DM患者因阻生拔除的外观完好无牙髓病症状的第三磨牙,对照组30例为正常人因阻生拔除的健康牙齿。采用酶联免疫吸附试验(ELISA)分别检测2组牙髓组织中IL-8的含量。光镜下观察2组牙髓组织的结构特点,并对牙髓标本进行血管内皮生长因子(VEGF)的免疫组化染色,通过Image pro-plus 6.0图像分析软件对牙髓组织中VEGF染色进行定量分析。应用SAS6.12软件包对试验数据进行统计分析。结果 DM组牙髓中IL-8含量高于正常牙髓(P=0.0455<0.05)。光镜下DM组牙髓中可见毛细血管壁增厚。VEGF在对照组和DM组牙髓中都有表达,DM组牙髓中的表达增强(P<0.01,成牙本质细胞P<0.001,成纤维细胞P<0.01,血管内皮细胞P<0.01)。结论 DM患者即使是无龋坏、无创伤缺损、无症状的牙齿牙髓也出现了炎症反应。VEGF在DM患者牙髓中的高表达提示其牙髓发生了损伤与修复过程,发生在DM患者外周微血管的病变同样也发生在了牙髓中,提示DM与其牙髓病变具有相关性。