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1.
The integrity of connective tissues surrounding dental implants may be influenced by a balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The purpose of this study was to provide an overall assessment of TIMP-1, MMP-1 and -8 levels as well as collagenase activities during the wound healing process after implantation and in peri-implantitis lesions. Peri-implant crevicular fluid (PICF) was sampled with sterile paper strips from 10 osseointegrated implants of 6 subjects. Ten implants from 6 patients affected with peri-implantitis were also assessed. Gingival crevicular fluid (GCF) from 11 periodontitis-affected patients and 10 healthy volunteers served as controls. TIMP-1 and MMP-1 and -8 protein levels in the PICF were measured by ELISA, and active and APMA-activatable collagenase activities were determined by functional assays using image-analysis after SDS-PAGE. The experiment showed a significant increase in the TIMP-1 level at 1 week after implantation as compared with that in GCF from healthy periodontium. Four weeks after implantation it had reached the same level as that in the GCF of healthy subjects. The data has also disclosed a higher post-implantation collagenase activity level at 1 week than at weeks 2, 4, and 12. This may be due to the increase in MMP-1 and -8. Furthermore, peri-implantitis and periodontitis were shown to be similar inflammatory lesions in respect to MMP-1 and -8 and collagenase activities, even though the TIMP-1/MMP-1 + MMP-8 ratio was significantly lower in peri-implantitis than in periodontitis. In conclusion, the overproduction of TIMP-1 in the wound area after implantation could, to some extent, inhibit excessive tissue destruction and degradation of the neo-matrix in wound repair due to MMPs.  相似文献   

2.
Nomura T  Ishii A  Oishi Y  Kohma H  Hara K 《Oral diseases》1998,4(4):231-240
OBJECTIVES: To provide an overall assessment of levels of tissue inhibitors of metalloproteinases (TIMPs), collagenase activities, and of immuno-reactivities for matrix metalloproteinases (MMP)-1 and -8 in gingival crevicular fluid (GCF) obtained from healthy subjects, and gingivitis and periodontitis patients, and to analyse the relationships between periodontal tissue destruction and the GCF components in periodontal diseases by principal component analysis. MATERIALS AND METHODS: GCF was sampled with sterile paper strips from 10 gingivitis and 11 periodontitis patients. Ten volunteers served as clinically healthy controls. TIMP-1 and -2 protein amounts in GCF were measured by ELISA, and active and APMA-activatable collagenase activities were determined by functional assays using image-analysis after SDS-PAGE. RESULTS: GCF TIMP-1 level and both active and latent collagenase activities were significantly higher in the diseased groups than in the healthy group. TIMP-2 was detectable in only 29% of all subjects (mean: 2.06 ng). Western blot analysis showed that MMP-8 was the major interstitial collagenase in the GCF of the diseased groups. Principal component analysis using clinical parameters and the GCF components has indicated components one to three account for 87% of total variation when evaluating the relevance of their measurements to periodontal diseases. CONCLUSIONS: We conducted the functional and immunological characterization of MMPs and TIMPs in the GCF of periodontally diseased patients. Principal component analysis indicated components one to three explaining 87% of total variation, and further suggested that higher collagenase activity (especially in active collagenase) would be an important marker in evaluating the pathogenesis of periodontitis. Consequently, these observations may have significant therapeutic and diagnostic implications.  相似文献   

3.
人类牙周膜组织中MMP-2及TIMP-2的比较研究   总被引:1,自引:0,他引:1  
目的:比较基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制因子-2(TIMP-2)在健康牙周组织(PDL)和病变牙周组织中的表达。方法:收集20例健康PDL和病变PDL组织样本,用ELISA法检测其中的MMP--2,TIMP-2水平。结果:牙周炎患者组PDL中的MMP-2的水平明显高于健康组(P〈0.01),牙周炎患者组PDL中的TIMP-2的水平高于健康组(P〈0.05),但MMP-2/TIMP-2明显增高。结论:研究提示MMP-2/TIMP-2失调与牙周组织破坏有关。  相似文献   

4.
The present study characterizes the periodontal status of patients with Sjögren's syndrome (SS) and measures collagenase, elastase and gelatinase in gingival crevicular fluid (GCF) from these patients compared with adult individuals with periodontitis and healthy controls. The periodontal status was assessed by the Gingival Bleeding Index (GBI), the Visible Plaque Index (VPI), and pocket depth. Activity measurements were performed for collagenase with SDS-PAGE/ laser densitometry, for elastase spectrophotometrically using a synthetic N-succinyl-Ala-Ala-Val p-nitroanilide peptide substrate, and for gelatinase with zymography. Seven of the 8 patients with SS and xerostomia showed a periodontium comparable to that seen in healthy controls. The GCF collagenase and elastase were significantly lower in patients with SS and in healthy controls than in patients with adult periodontitis. It was noteworthy that the one SS patient with periodontitis had high GCF collagenase and elastase activity. In all study groups multiple forms of gelatinases were present, indicating that they represent constitutively expressed proteinases involved in normal tissue remodeling processes. Our findings suggest that periodontal pockets/GCF form a micromilieu not affected by involvement of glandular tissue and, therefore, patients with SS show, clinically and biochemically, a periodontal status comparable to that seen in healthy controls.  相似文献   

5.
目的:探讨NF-κB通路对牵张应力介导的人牙周膜细胞(HPDLCs)中MMP-2表达的影响.方法:按胶原酶消化法培养HPDLCs,分为5组:非加力对照组(A组);12%形变率,3h组(B组);12%形变率,6h组(C组);12%形变率,12h组(D组);12%形变率+PDTC干预,12h组(E组),通过细胞牵张应力加载系统施加机械牵张应力,结束后收集细胞提取蛋白,用蛋白印迹法检测MMP-2蛋白表达的变化.结果:B、C、D组MMP-2表达大于A组,差异有统计学意义(P<0.05),且随着作用时间的延长蛋白表达增加;NF-κB通路抑制剂PDTC可抑制机械牵张应力作用下牙周膜细胞MMP-2表达的增加.结论:持续机械牵张应力可诱导人牙周膜细胞MMP-2表达的增加,从而影响牙周组织细胞外基质代谢,机械牵张应力可通过NF-κB通路影响MMP-2蛋白的表达.  相似文献   

6.
Total TIMP-1 concentration in whole saliva of periodontally diseased subjects, 137 ± 67 ng/ml (mean ± SD), was clearly lower (p < 0.001) than that of clinically healthy subjects, 273±145, and that of edentulous subjects, 332±121. On the contrary, both active [1.58±0.35 units/ml (mean ± SD)] and total (2.08 ± 0.25) collagenase activities inTIMP-1-free whole saliva of diseased subjects were significantly higher than the activities (0.14 ± 0.14 and 0.50 ± 0.27, respectively) in TIMP-1-free whole saliva of healthy subjects. Most of the total collagenase in whole saliva of healthy subjects consisted of procollagenase, while mainly active collagenase was present in whole saliva from patients with periodontal diseases. Significant reciprocal changes of TIMP-1 and collagenase levels, that is, increase in TIMP-1 concentration and decrease in collagenase activity, were observed after the initial therapy of periodontitis patients.  相似文献   

7.
Orthodontic force induces biochemical responses in the periodontal ligament (PDL), but the matrix metalloproteinase (MMP)-dependent molecular mechanisms in orthodontically induced periodontal remodeling have remained unclear. Previous studies indicate that mechanical stress induces MMP-1 production in human PDL cells in vitro. We tested the hypothesis whether the in vivo levels, molecular forms, and degree of activation of MMP-1 and MMP-8 in gingival crevicular fluid (GCF) reflect an early stage of orthodontic tooth movement. Molecular forms of MMP-1 and MMP-8 were analyzed by Western blot, and MMP-8 levels by quantitative immunofluorometric assay (IFMA). The results showed that GCF MMP-8 levels for orthodontically treated teeth were significantly higher at 4-8 hrs after force application than before activation, and when compared with the control teeth (p < 0.05). Analysis of our data indicates that the cells within the periodontium are up-regulated to produce MMP-8, and the increased expression and activation of GCF MMP-8 reflect enhanced periodontal remodeling induced by orthodontic force.  相似文献   

8.
BACKGROUND: Matrix metalloproteinases (MMPs) process the extracellular matrix and act in tissue remodelling in many physiological and pathological conditions. Certain MMPs can also exert protective anti-inflammatory properties. The levels and expression of MMPs and tissue inhibitors of MMPs (TIMPs) in saliva and gingival tissues of human immunodeficiency virus-seropositive (HIV+) patients are unclear. METHODS: Enzyme-linked immunosorbent assay methods and Western blots were used to study levels and molecular forms of MMP-7, -8, -9, -25, and -26 and TIMP-1 from salivary samples of HIV+ patients (n = 55) and healthy controls (n = 10). The expression of MMPs was also studied by immunohistochemical means in gingival tissue specimens (n = 11, HIV+ patients; n = 10, healthy controls). RESULTS: The HIV+ patients' MMP-8 levels in saliva were statistically significantly higher only in the acquired immunodeficiency syndrome (AIDS)-phase. MMP-9 levels in ASX- and AIDS-phases showed increased expression. TIMP-1 levels were significantly decreased in lymphadenopathy syndrome (LAS)- and AIDS-related complex (ARC)-phases, while MMP-8/TIMP-1 and MMP-9/TIMP-1 molar ratios were increased in all phases in comparison with controls. The molecular forms of MMP-7, -25, and -26 were different between patients and controls as assessed by Western blot. Immunohistochemical studies showed slightly enhanced MMP-7, -8, -9, -25, and -26 staining in HIV+ gingival tissue samples in comparison with controls. CONCLUSIONS: This study confirmed and further demonstrated differences in salivary amounts and molecular forms of MMPs and TIMP-1 in HIV+ patients. The results may reflect alterations in host defence reactions associated with HIV infection.  相似文献   

9.
Abstract The aim of the present study was to characterize the eventual presence and molecular forms of gelatinase/type IV collagenase activities in gingival crevicular fluid (GCF) and saliva in different forms of periodontitis: patients with clinically healthy periodontiura served as controls. Enzyme activities were monitored electrophoretically by zymography using gelatin and type IV collagen as substrates and analyzed visually and/or densitometrically. Both saliva and GCF collected from adult periodontitis, localized juvenile periodontitis and type II diabetes mellitus periodontitis patients contained species moving identically with gelatinase isolated from human neutrophils or MMP-9 (mean 98 kD), and species with mobility similar to gelatinase in fibroblast cell culture supernatants or MMP-2 (mean 76 kD). Hitherto, undescribed high molecular weight forms (mean 128 kD), were found, possibly representing polymerized or complexed enzyme active/activated in situ in the gel matrix. Small molecular forms of gelatinases (mean 51 kD and 46 kD), unable to cleave type IV collagen, were also found, most likely representing in vivo proteolytically activated, truncated enzymes. Although multiple forms of gelatinases/type IV collagenases in saliva and GCF may take part in the tissue destruction in periodontitis, their profile judged according to molecular weights does not differentiate between different forms of periodontitis.  相似文献   

10.
Saliva collected from subjects with healthy and with diseased periodontium was assayed for collagenase activity by incubation at 25 degrees C with soluble type I, II or III collagen. The degradation products were analyzed by separation in SDS-polyacrylamide gel electrophoresis followed either by protein staining or by exposure of the dried gel to X-ray film in the case of radioactively labeled type I collagen. Collagenase of vertebrate type was detected in the whole saliva of all subjects but not in parotid, sublingual or submandibular fluids. Most of the collagenase was in the soluble fraction of saliva that also contained factors which both activated and inhibited the enzyme. The salivary collagenase resembled the collagenase of human PMNs and gingival sulcular fluid in its molecular size of 70,000 daltons, in its activation by gold thioglucose and in its tendency to degrade types I and II collagens over type III collagen. Before periodontal treatment, the saliva of periodontitis patients had significantly higher collagenase than after treatment. In periodontitis, collagenase existed mainly in the active form, while in the healthy mouths most of the enzyme was latent but could be activated by sulfhydryl reagents or proteolytically with trypsin, and chymotrypsin but not by human plasma kallikrein or plasmin. In some of the samples from untreated periodontitis patients bacterial collagenase may have been present in small quantities. Most of the collagenase in the saliva from all subjects appeared to originate from PMNs entering the oral cavity through the gingival sulcus.  相似文献   

11.
BACKGROUND AND OBJECTIVE: The aim of this work was to investigate periodontal status, in relation to inflammatory markers and cortisol, in the gingival crevicular fluid and saliva of a homogenous group of women on long-term sick leave for job-stress related depression in comparison to nondepressed women. MATERIAL AND METHODS: The participants comprised 20 women with depression (DSM-IV) (mean age 48.5 +/- 6.9 years) and 29 healthy controls (mean age 54.5 +/- 2.9 years). Clinical examination was performed. Gingival crevicular fluid was collected by an intracrevicular washing technique. Interleukin-1 beta, interleukin-6, matrix metalloproteinase (MMP)-8 and MMP-9 were determined with enzyme-linked immunosorbent assay and cortisol was determined by using a radioimmunoassay. One-way analysis of covariance was used as the statistical method. RESULTS: The depressed patients had significantly higher gingival inflammation (p < 0.001), and deeper pockets (p < 0.003), than the healthy controls, after adjusting for age and smoking. The levels of interleukin-6 in the gingival crevicular fluid were significantly higher in the patients than in the controls: 3.84 +/- 1.58 pg per site and 0.79 +/- 1.83 pg per site, respectively, p < 0.003. There were no significant differences in the levels of interleukin-1 beta, MMP-8 and MMP-9. The patients had lower cortisol values in gingival crevicular fluid than the controls, whereas the levels of cortisol in saliva were similar in both groups. CONCLUSION: Women on long-term sick-leave for depression had more severe periodontitis and higher concentrations of interleukin-6 in gingival crevicular fluid than healthy controls. An alteration of the immune system in these patients might be interpreted as reflecting the consequences of long-term stress exposure and might contribute to worse periodontal conditions in these particular patients.  相似文献   

12.
13.
Eight adult periodontitis (AP) patients were studied immunohistochemically to determine the presence of matrix metalloproteinases (MMPs) MMP-1, MMP-3, and MMP-8 in the marginal gingival and gingival granulation tissue specimens obtained from periodontal flap surgery after scaling and root planing. Clinically healthy gingival tissue specimens obtained from impacted third-molar extraction operations served as controls. MMP-type-specific antisera were applied by the avidin-biotin-peroxidase complex staining method. Moderate immunoreactivity for neutrophil collagenase (MMP-8) was found both in the AP patients' marginal gingival connective tissue and in gingival granulation tissue specimens. Immunoreactivity for fibroblast-type collagenase (MMP-1) and stromelysin-1 (MMP-3) was detected only in the AP patients' gingival granulation tissue specimens. In the control specimens, no immunoreactivity for the MMPs could be detected. For the first time, this finding demonstrates immunohistochemically the presence of MMP-8 in human inflamed gingiva in situ, and further highlights the importance of MMP-8 in periodontal tissue destruction, evidently during the acute phase(s) of the disease. However, our results confirm and extend previous studies indicating that other types of MMPs from resident gingival cell sources also seem to participate in the chronic and destructive course of periodontal inflammation.  相似文献   

14.
Abstract Matrix metalloproteinases (MMPs) and serine proteinases seem to be related to tissue destruction in periodontitis. The presence of MMPs in gingival crevicular fluid (GCF) and saliva, however, has not been studied comprehensively with the enzyme-linked immunosorbent assay (ELISA)-technique, We therefore examined the levels of MMP-1,-3.-8 and -9, and their endogenous inhibitor, tissue inhibitor of matrix metalloproteinases (TIMP-1). in GCF and saliva of patients with adult periodontitiss (AP) and localized juvenile periodontitis (LJP). Elevated levels of MMP-1 were detected in LJP GCF compared to AP and control GCF. Elevated levels of TIMP-1 were also detected in LJP GCF in comparison to AP and control GCF Higher MMP-8 levels were detected in AP GCF compared to LJP and control GCF. The relative low levels of MMP-3 were present in all studied GCF samples. Elevated levels of MMP-8 were further detected in saliva of AP compared to LJP and the controls. Both MMP-1 and TIMP-1 were detected in all studied saliva samples, but not significant differences were detected between the studied groups. Our ELISA-results confirm that (i) PMN MMP-8 and MMP-9 are the main collagenase and gelatinase in AP GCF, whereas GCF collagenase in LJP seems to be of the MMP-1-type; (ii) only low levels of TIMP-1. endogenous MMP-inhibitor. are present in AP GCF. which emphasises the importance of doxycycline as a possible adjunctive drug in the treatment of AP patients; (iii) tests based on specific antibodies against PMN MMPs. especially MMP-8, might serve as a reliable method of measuring and monitoring enzyme levels in GCF from different periodontitis patients.  相似文献   

15.
We studied the salivary levels and activities of the matrix metalloproteinases (MMP) -8 and -9 in 45 type 2 diabetic patients and 77 control subjects. The patients' mean glycosylated haemoglobin (HbA1c) was 8.7%, indicating an unsatisfactory metabolic control of the disease. The MMP levels were further related to the clinical and microbiological periodontal findings as well as to salivary flow rate and other factors. The salivary flow rate, albumin and amylase concentrations were similar in type 2 diabetic patients to those in the control group. The mean gingival and periodontal pocket indexes were higher in the diabetes group. The number of potential periodontopathogenic bacteria was lower, however, in the diabetic than in the control group. Zymography and immunoblotting revealed that the major MMPs in the type 2 diabetic patients' saliva were MMP-8 and MMP-9. Salivary MMP levels and activities in type 2 diabetic patients were in general similar to those in the control group. However, the correlation coefficients using multiple regression analysis revealed that gingival bleeding, pocket depths and HbA1c were associated with increased MMP-8 levels which, in turn, were negatively predicted by elevated plasma lipid peroxide levels in the diabetic group. Our data on salivary MMP-8 and -9 do not support the concept of generalized neutrophil dysfunction in unbalanced diabetes. Moreover, plasma lipid peroxidation levels reflecting the increased oxidative burden, which is generated mainly by triggered neutrophils, do not indicate neutrophil dysfunction due to diabetes, but may rather be related to the increased tissue damage in an uncontrolled disease. However. advanced periodontitis in type 2 diabetes seems to be related to elevated salivary MMP-8 levels which might be useful in monitoring periodontal disease in diabetes.  相似文献   

16.
Human neutrophil-type (MMP-8) and fibroblast-type (MMP-1) interstitial collagenase, and their inhibition by tetracyclines in saliva from patients with recurrent aphthous ulcers (RAU) or aphthae, were studied by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and enzymological analyses. In the salivary specimens obtained from patients with aphthae, collagenase was found in endogenously active form and was predominantly of MMP-8 type. Topical rinsing treatment with chlortetracycline (Aureomycin®) alleviated the discomfort caused by the lesions but did not reduce salivary collagenase amounts; however in vitro , doxycycline inhibited salivary collagenase totally.  相似文献   

17.
Abstract Periodontal diseases may be the first clinical sign of human immunodeficiency virus (HIV)-infection. Since the immunosuppression and subsequent susceptibility may alter the responses of the oral tissues as well as the microflora, both periodontal treatment and result of therapy may be modified. The periodontal diseases in HIV-seropositive patients include common as well as less conventional forms of gingivitis and periodontitis, and bacteria, mycotic and viral infections are seen. Neoplasias may also involve the periodontium; most common are Kaposi's sarcoma and non-Hodgkin's lymphoma. Recent studies of unselected groups of patients indicate that periodontal health in at least some groups of HIV-seropositive patients is better than previously reported.  相似文献   

18.
BACKGROUND: The aim of this work was to improve the assessment of the periodontal disease status through measurements of extracellular matrix metalloproteinases (MMPs) and their tissular inhibitors (TIMPs) in the gingival crevicular fluid from patients diagnosed with chronic periodontitis. METHODS: Gingival crevicular fluid samples from patients (n = 13) were taken from 60 sites initially, and from 51 and 41 sites, respectively, 3 and 6 months after scaling and root planing. Gingival crevicular fluid samples were also taken from healthy subjects (n = 11, 24 sites). The presence of MMP-9 and MMP-8 was assessed by zymography and immunowestern blotting, respectively. The actual MMP activity (gelatinase and collagenase) was measured using the fluorogenic substrate assay. TIMP-1 and -2 levels were measured by immunodot blot. RESULTS: The fluorogenic substrate assay determinations showed higher MMP activity in sites with probing depth > or = 4 mm, with significant reduction post-treatment. Gelatinase activity followed by zymography consisted mainly of MMP-9. A different pattern of MMP-8 in control and patient sites was found. Controls only showed species of a partially active form (69 kDa), whereas patient sites showed a high frequency of the active form (56 kDa), and in some cases the latent form (85 kDa) was also observed. The active form reduced its frequency in sites with probing depth > or = 4 mm. TIMP-1 and -2 levels in patients were significantly lower than in controls, and after treatment the recovery of TIMP-1 level similar to control was observed. CONCLUSION: Significant correlations between the severity of the periodontal disease and the actual MMP activity, the active form of MMP-8 and the low level of both TIMP-1 and TIMP-2 were found.  相似文献   

19.
The introduction of highly active antiretroviral therapy (HAART) has decreased the incidence and prevalence of several oral manifestations such as oral candidiasis, hairy leukoplakia, and Kaposi's sarcoma in HIV-infected patients. Regarding periodontal disease the findings are not clear. This disease represents a group of chronic oral diseases characterized by infection and inflammation of the periodontal tissues. These tissues surround the teeth and provide periodontal protection (the gingival tissue) and periodontal support (periodontal ligament, root cementum, alveolar bone). Clinical, immunological, and microbiological aspects of these diseases, such as linear gingival erythema (LGE), necrotizing periodontal diseases (NPD) (necrotizing ulcerative gingivitis [NUG], necrotizing ulcerative periodontitis [NUP] and necrotizing stomatitis), and chronic periodontitis, have been widely studied in HIV-infected individuals, but without providing conclusive results. The purpose of this review was to contribute to a better overall understanding of the probable impact of HIV-infection on the characteristics of periodontal infections.  相似文献   

20.
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.  相似文献   

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