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.     

Involvement of MT1‐MMP and TIMP‐2 in human periodontal disease

Oral Diseases (2010) 16 , 388–395 Objectives: Periodontal disease is characterized by an increased collagen metabolism. Although membrane type‐1 matrix metalloproteinase (MT1‐MMP) plays a critical role in collagen degradation, its involvement in human periodontitis remains to be determined. Methods: MT1‐MMP and TIMP‐2 expression and distribution were evaluated in gingival tissue samples derived from 10 healthy and 12 periodontitis‐affected human subjects. MT1‐MMP and TIMP‐2 expression were assessed through Western‐blot of tissue homogenates. The main cell types involved in MT1‐MMP and TIMP‐2 production were evaluated by means of immunohistochemistry. Results: Both MT1‐MMP and TIMP‐2 were significantly increased in periodontitis‐affected gingival tissues when compared to healthy gingiva. Moreover, the balance between MT1‐MMP and its inhibitor TIMP‐2 was altered in periodontitis‐affected tissues, suggesting an imbalance in this proteolytic axis. Immunohistochemistry demonstrated the expression of MT1‐MMP in fibroblasts and macrophages in gingival tissues. MT1‐MMP was detected in cells in close association with the gingival collagen matrix. TIMP‐2 expression was identified in fibroblasts, macrophages and epithelial cells. Conclusions: Our observations show an increased expression of MT1‐MMP and TIMP‐2 in periodontitis‐affected gingival tissues. The altered balance between these two molecular mediators of collagen remodeling suggests their involvement in human periodontal disease.     

Matrix Metalloproteinase (MMP)‐8 and Tissue Inhibitor of MMP‐1 (TIMP‐1) Gene Polymorphisms in Generalized Aggressive Periodontitis: Gingival Crevicular Fluid MMP‐8 and TIMP‐1 Levels and Outcome of Periodontal Therapy

Background: The aim of the present study is to investigate matrix metalloproteinase (MMP)‐8 and tissue inhibitor of MMP‐1 (TIMP‐1) gene polymorphisms in generalized aggressive periodontitis (GAgP) and to assess the effects of MMP‐8 and TIMP‐1 genotypes on the outcomes of non‐surgical periodontal therapy. Methods: Genomic DNA was obtained from peripheral blood of 100 patients with GAgP and 167 periodontally healthy controls. MMP‐8 +17 C/G, ?799 C/T, ?381 A/G and TIMP‐1 372 T/C, *429 T/G polymorphisms were determined by polymerase chain reaction‐restriction fragment length polymorphism. Patients with GAgP received non‐surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and gingival crevicular fluid (GCF) samples were collected at baseline and at follow‐up visits. GCF biomarkers were analyzed by immunofluorescence assay and enzyme‐linked immunosorbent assay. Results: Distribution of the MMP‐8 ?799 C/T genotypes was significantly different between the GAgP and control groups (P <0.005). TIMP‐1 372 T/C and *429 T/G genotypes in males were also significantly different between study groups (P <0.004). GCF MMP‐8 levels decreased until 3 months after non‐surgical therapy compared with baseline in T and G alleles, as well as G and C allele carriers (P <0.0125), whereas no significant decreased was observed in non‐carriers (P >0.0125). Conclusion: On the basis of the present findings, it can be suggested that MMP‐8 ?799 C/T and TIMP‐1 372 T/C, *429 T/G gene polymorphisms in males may be associated with the susceptibility to GAgP in the Turkish population.     

MMP-13 and TIMP-1 determinations in progressive chronic periodontitis

Matrix metalloproteinase (MMP)-13 is a collagenase involved in extracellular matrix degradation either by its direct degradative effects or by processing bioactive substrates. The aim of this study was to determine the levels of MMP-13 and tissue inhibitor of metalloproteinase (TIMP)-1 in gingival crevicular fluid (GCF) and gingival biopsies obtained from active and inactive sites during chronic periodontitis progression. MATERIALS AND METHODS: This was a longitudinal study in which chronic periodontitis patients with moderate to severe disease were included and followed until they developed progression determined by the tolerance method. GCF samples were obtained from periodontitis, active, inactive and healthy sites and additional gingival biopsies were taken from active and inactive sites. MMP-13 and TIMP-1 determinations were carried out by immunodot blots and immunowestern blots. RESULTS: In progressive periodontitis, MMP-13 and TIMP-1 remained unchanged between active and inactive sites, but as the TIMP-1 relative levels increased together with MMP-13 elevation in inactive samples, an inverse correlation was observed in active sites. Besides, MMP-13 was undetectable in healthy controls. CONCLUSION: Chronic periodontitis is characterized by increased MMP-13 expression. During disease progression, active sites tended to decrease TIMP-1 levels in association with MMP-13 elevation.     

Analysis of matrix metalloproteinase (MMP‐8 and MMP‐2) activity in gingival crevicular fluid from children with Down’s syndrome

Yamazaki‐Kubota T, Miyamoto M, Sano Y, Kusumoto M, Yonezu T, Sugita K, Okuda K, Yakushiji M, Ishihara K. Analysis of matrix metalloproteinase (MMP‐8 and MMP‐2) activity in gingival crevicular fluid from children with Down’s syndrome. J Periodont Res 2010; doi: 10.1111/j.1600‐0765.2009.01214.x. © 2009 John Wiley & Sons A/S Background and Objective: High levels of colonization by periodontopathic bacteria and a high prevalence of chronic inflammatory periodontal disease have been reported in children with Down’s syndrome. Matrix metalloproteinases (MMPs) are mediators of extracellular matrix degradation and remodelling, and are deeply involved in the course of periodontal disease. To clarify the relationship between Down’s syndrome and periodontitis, we investigated levels of MMP‐2 and MMP‐8 in gingival crevicular fluid (GCF) and detection of periodontopathic bacteria from subgingival plaque. Material and Methods: Samples of GCF and plaque were isolated from central incisors. Levels of MMPs were evaluated by enzyme‐linked immunosorbent assay, and periodontopathic bacteria were detected by polymerase chain reaction. Results: Levels of MMP‐2 and MMP‐8 in Down’s syndrome patients were higher than those in healthy control subjects. In the Down’s syndrome group, increases in these MMPs were observed in GCF from patients with an oral hygiene index score of < 2 and in GCF from sites that were negative for bleeding on probing. The detection rate of periodontopathic bacteria in Down’s syndrome patients was higher than that in the control subjects. Matrix metalloproteinase‐2 levels in sites harbouring Porphyromonas gingivalis or Aggregatibacter (Actinobacillus) actinomycetemcomitans were lower than in those without these microorganisms. Conclusion: These results suggest an increase in MMP‐2 and MMP‐8 in Down’s syndrome patients, regardless of whether inflammation of periodontal tissue is present or not.     

Oral rinse MMP‐8 point‐of‐care immuno test identifies patients with strong periodontal inflammatory burden

Oral Diseases (2010) 17 , 115–122 Objective: To determine whether oral rinse matrix metalloproteinase (MMP)‐8 levels, measured by three different methods, tissue inhibitor of matrix metalloprotease‐1 (TIMP‐1) levels and elastase activity differentiate subjects with different periodontal condition; and second, to find out if MMP‐8 levels were comparable among the methods used. Methods: MMP‐8 levels were analysed with an immunofluorometric method (IFMA), dentoELISA and commercial ELISA. Also TIMP‐1 levels and elastase activity were measured. For statistical analysis 214 study subjects were categorized into four groups, specified by the presence and number of moderate (4–5 mm) and deep (≥6 mm) periodontal pockets, and bleeding on probing percentage. Results: MMP‐8 levels especially measured by dentoELISA and adjusted to the number of teeth per subject differentiated the study group with strong periodontal inflammatory burden from groups with lower levels. This was also verified with receiver operating characteristic ( ROC) analysis. Elastase activity associated with higher IFMA and dentoELISA MMP‐8 levels. IFMA MMP‐8/TIMP and dentoELISA MMP‐8/TIMP‐1 tended to be higher with the increasing level of periodontal inflammatory burden. TIMP‐1 levels decreased with increasing age. Conclusions: Oral rinse MMP‐8 together with TIMP‐1 analysis may have potential in complementary periodontal diagnostics. dentoELISA can be applied in quantitative oral rinse chair side biomarker diagnostics.     

Matrix metalloproteinase‐8 concentration in shallow crevices associated with the extent of periodontal disease

Objective: The aim of this study was to analyse the association between matrix metalloproteinase‐8 (MMP‐8) concentration in shallow, mostly non‐bleeding gingival crevices, and the extent of periodontal disease. Material and Methods: Plaque, bleeding on probing (BOP), probing pocket depth (PPD) and attachment level (AL) were assessed clinically in 48 patients with chronic periodontitis. MMP‐8 concentrations in gingival crevicular fluid (GCF) from four shallow (PPD3 mm), and four diseased sites and in serum, were measured by enzyme‐linked immunosorbent assay. Results: The mean concentration of MMP‐8 in GCF from shallow crevices was 11.8 ± 12.8 ng/ml and from diseased sites was 150.1 ± 91.8 ng/ml. In subjects with moderate to high plaque scores, a statistically significant association was found between MMP‐8 concentration from shallow crevices and the extent of AL4 mm (p=0.028) and AL6 mm (p<0.001). Conclusion: The above association between MMP‐8 concentration in shallow crevices and attachment loss provides a new aspect to future studies of MMP‐8 as a prognostic marker for periodontal disease.     

The effect of smoking on the mRNA expression of MMPs and TIMP-1 in untreated chronic periodontitis patients: a cross-sectional study

Mouzakiti E, Pepelassi E, Fanourakis G, Markopoulou C, Tseleni‐Balafouta S, Vrotsos I. The effect of smoking on the mRNA expression of MMPs and TIMP‐1 in untreated chronic periodontitis patients: a cross‐sectional study. J Periodont Res 2011; 46: 576–583.©2011 John Wiley & Sons A/S Background and Objective: Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are important for extracellular matrix. Expression of MMPs has been evaluated in gingiva without studying smoking. The aim of this study was to explore the effect of smoking on mRNA expression of MMP‐1, ‐3, ‐8, ‐9 and ‐13 and TIMP‐1 in untreated chronic periodontitis and in periodontal health. Material and Methods: Gingival samples were harvested from 30 subjects with untreated chronic periodontitis (15 nonsmokers and 15 smokers) and 30 periodontally healthy subjects (15 nonsmokers and 15 smokers). Full‐mouth plaque score, gingival index, bleeding on probing, probing depth and clinical attachment level were recorded. Total RNA was isolated, and the mRNA expression of MMPs and TIMP‐1 was assessed by RT‐PCR. Results: Periodontitis groups were comparable in clinical measurements. Nonsmoker subjects with periodontitis had statistically significantly higher MMP‐1, lower MMP‐9 and TIMP‐1 expression and higher MMP‐1/TIMP‐1 ratio than smokers; and higher MMP‐8 expression and MMP‐8/TIMP‐1 and MMP‐1/TIMP‐1 ratios than healthy nonsmokers. Healthy nonsmokers had statistically significantly higher MMP‐13 expression than healthy smokers. Smoker periodontitis and healthy subjects had similar expression levels of MMPs and TIMP‐1 and MMPs/TIMP‐1 ratios. There was correlation among the MMPs only for smoker periodontitis subjects. Expression of MMP‐13 was correlated with mean clinical attachment level. Conclusion: Within its limits, this study demonstrated that smoking affected mRNA expression of MMPs and TIMP‐1, MMPs/TIMP‐1 ratios and relationships among MMPs in untreated chronic periodontitis and expression of MMPs in health. In the absence of smoking, chronic periodontitis affected expression of MMPs and MMPs/TIMP‐1 ratios.     

Nicotine modulates gelatinase B (MMP‐9) and epilysin (MMP‐28) expression in reconstituted human oral epithelium

J Oral Pathol Med (2011) 40 : 33–36 Oral epithelial keratinocytes express nicotinic cholinergic receptors which activation modulates keratinocytes differentiation and migration through different metabolic pathways. Matrix metalloproteinases (MMPs) are Zn‐dependent enzyme involved in cell migration. Among them, gelatinase B (MMP‐9) and epilysin (MMP‐28) are two MMPs expressed by human keratinocytes during both wound healing and proliferation. Their expression has been investigated in a reconstituted human oral epithelium (HOE) exposed to nicotine (Nic, 1–50 μM) for 72 h both in the absence and presence of the nicotinic antagonist mecamylamine (Mec), H7, a PKC inhibitor and PD98059, a MAPK inhibitor (PD). At the end of treatment, MMP‐28 expression has been analyzed in epithelium sections using an anti‐MMP‐28 antibody, whereas MMP‐9 presence and activity has been measured in cell‐conditioned medium analyzed by gelatine zymography. The expression of MMP‐9 was reduced by Nic in a dose‐dependent fashion and this effect was antagonized by Mec, H7 and PD. On the other hand, Nic increased the expression of MMP‐28, and this effect was blocked both by H7 and PD, whereas Mec even enforced it. Nic effects on MMP‐9 and MMP‐28 expression by oral keratinocytes were not previously reported and these data suggest MMPs expression mediated by PKC and MAPK as a possible target for Nic toxicity in oral epithelium.     

Expression of MMPs and TIMP-1 in smoker and nonsmoker chronic periodontitis patients before and after periodontal treatment

Mouzakiti E, Pepelassi E, Fanourakis G, Markopoulou C, Tseleni‐Balafouta S, Vrotsos I. Expression of MMPs and TIMP‐1 in smoker and nonsmoker chronic periodontitis patients before and after periodontal treatment. J Periodont Res 2012; 47: 532–542. © 2012 John Wiley & Sons A/S Background and Objective: Nonsurgical periodontal treatment controls periodontal inflammation. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are implicated both in the destruction and in the healing of periodontal tissues. The aim of the present study was to compare the mRNA expression of MMP‐1, ‐3, ‐8, ‐9 and ‐13 and TIMP‐1 in chronic periodontitis before and after initial periodontal treatment. Material and Methods: Ninety gingival samples were harvested from 30 patients with chronic periodontitis (15 nonsmokers and 15 smokers) before and after nonsurgical treatment and from 30 periodontally healthy control subjects (15 nonsmokers and 15 smokers). Clinical parameters were assessed before and after treatment. Total RNA was isolated, and mRNA expression of MMPs and TIMP‐1 was assessed by RT‐PCR. Results: Periodontal treatment significantly increased TIMP‐1 expression and decreased the ratios of MMPs/TIMP‐1. Post‐treatment, nonsmokers with periodontitis had significantly higher MMP‐8 and TIMP‐1 expression than healthy nonsmokers, and smokers with periodontitis had significantly higher MMP‐13 and TIMP‐1 expressions than healthy smokers. Post‐treatment, smokers had significantly higher TIMP‐1 expression and lower MMP‐8/TIMP‐1 ratio than nonsmokers. Post‐treatment, there was no correlation among MMPs, and the expression of MMPs and TIMP‐1 was not correlated with clinical measurements. Conclusion: Periodontal treatment increased TIMP‐1 expression and decreased the ratios of MMPs/TIMP‐1 in chronic periodontitis. The post‐treatment increase in TIMP‐1 expression was higher for smokers. The TIMP‐1 expression was higher post‐treatment than in health. Post‐treatment, MMP‐8 expression was higher in nonsmokers with periodontitis than in healthy nonsmokers, whereas MMP‐13 expression was higher in smokers with periodontitis than in healthy smokers.     

Gingival crevicular fluid and serum matrix metalloproteinase-8 and tissue inhibitor of matrix metalloproteinase-1 levels in renal transplant patients undergoing different immunosuppressive therapy

Aim: We investigated gingival crevicular fluid (GCF) and serum matrix metalloproteinase‐8 (MMP‐8) and tissue inhibitor of matrix metalloproteinase‐1 (TIMP‐1) levels from renal transplant patients receiving cyclosporine‐A (CsA) and having gingival overgrowth (GO), from patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy. Material and Methods: GCF samples were collected from sites with GO (GO+) and without GO (GO?) in CsA patients having GO; and GO? sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum MMP‐8 and TIMP‐1 levels were determined by a time‐resolved immunofluorometric assay (IFMA) and enzyme‐linked immunosorbent assay. Results: GO+ sites in CsA patients having GO had elevated GCF MMP‐8 levels compared with those of CsA patients having no GO, tacrolimus and healthy subjects (p<0.005), but these levels were similar to those of gingivitis. The GCF MMP‐8 level was higher in GO+ compared with GO? sites in CsA patients having GO (p<0.05). GCF TIMP‐1 levels were similar between groups. Tacrolimus patients had lower GCF MMP‐8 levels than gingivitis (p<0.005), but levels similar to the healthy group. Conclusion: These results show that CsA and tacrolimus therapy has no significant effect on GCF MMP‐8 levels, and gingival inflammation seems to be the main reason for their elevations.     

Effect of MMP-1 promoter polymorphisms on GCF MMP-1 levels and outcome of periodontal therapy in patients with severe chronic periodontitis

Aims: The aims of this study were to investigate (1) the matrix metalloproteinase‐1 (MMP‐1) promoter polymorphisms in severe chronic periodontitis (CP), (2) the relationship of periodontal therapy outcome with these genotypes, and (3) the gingival crevicular fluid (GCF) MMP‐1 levels–MMP‐1 genotype correlation. Material and Methods: Genomic DNA was obtained from the peripheral blood of 102 patients with severe CP and 98 periodontally healthy subjects. MMP‐1 ?519A/G and ?1607 1G/2G polymorphisms were determined by the polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method. Fifty‐eight CP patients received non‐surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and GCF samples were collected at baseline and at 6 months. GCF MMP‐1 levels were analysed by enzyme‐linked immunosorbent assay (ELISA). Results: The distribution of MMP‐1 genotypes did not significantly differ between the study groups. On the other hand, the ?1607 2G allele frequency of severe CP patients was higher than that of healthy subjects. MMP‐1 ?519G allele carriers had higher GCF MMP‐1 levels and percentage of sites with 4–6 mm clinical attachment level (CAL) compared with AA genotypes after non‐surgical periodontal therapy (p<0.05). Conclusions: These data suggest that the ?1607 2G polymorphic allele of the MMP‐1 gene could be associated with susceptibility to severe CP in the Turkish population. It seems that ?519AG and GG genotypes could play a role in the outcome of periodontal therapy.     

Origin of Galactose‐Deficient Immunoglobulin G in Gingival Crevicular Fluid in Periodontitis

Background: Periodontitis is a chronic inflammatory disease initiated by a synergistic and dysbiotic microbial community that elicits a gingival inflammatory response leading to tissue breakdown. Periodontitis shares many characteristics with other chronic inflammatory diseases, including abnormal glycosylation of immunoglobulin (Ig)G. The current authors have previously demonstrated that IgG from gingival crevicular fluid (GCF) of patients with chronic periodontitis contains galactose (Gal)‐deficient IgG. Methods: The origin of the aberrantly glycosylated IgG was determined by measuring levels of Gal‐deficient IgG in GCF and serum from patients with periodontitis and non‐periodontitis controls using lectin enzyme‐linked immunosorbent assay. The Ig‐producing cells and the proportion of cells producing Gal‐deficient IgG were immunohistochemically determined in gingival tissues from patients with periodontitis by fluorescence microscopy. The results were statistically evaluated and correlated with clinical data. Results: The results indicate that GCF of patients with periodontitis had higher levels of Gal‐deficient IgG compared with controls (P = 0.002). In gingival tissues, IgG was the dominant isotype among Ig‐producing cells, and 60% of IgG‐positive cells produced Gal‐deficient IgG. Moreover, the proportion of Gal‐deficient IgG‐producing cells directly correlated with clinical parameters of probing depth and clinical attachment loss (AL). Conclusion: These results suggest that the presence of Gal‐deficient IgG is associated with gingival inflammation and may play a role in the worsening of clinical parameters of periodontitis, such as AL.     

Smoking and matrix metalloproteinases,neutrophil elastase and myeloperoxidase in chronic periodontitis

Oral Diseases (2010) 17 , 68–76 Objectives: To investigate possible relationship between smoking and serum concentrations of matrix metalloproteinase‐8,‐9 (MMP‐8, MMP‐9), tissue inhibitor of matrix metalloproteinases‐1 (TIMP‐1), neutrophil elastase (NE), myeloperoxidase (MPO) in chronic periodontitis (CP) patients relative to periodontally healthy subjects. Methods: Serum samples were obtained from 111 subjects before initiation of any periodontal intervention. Fifty‐five CP patients (39 non‐smokers, 16 smokers) and 56 periodontally healthy subjects (39 non‐smokers, 17 smokers) were recruited. Serum concentrations of MMP‐8 were determined by IFMA and MPO, MMP‐9, TIMP‐1, NE concentrations by ELISA. ANCOVA and Pearson correlation analysis was utilized for statistical analysis. Results: Serum MPO, NE concentrations were higher in smoker CP than non‐smoker CP patients (P = 0.002 and P < 0.001, respectively), whereas these were similar in smoker, non‐smoker periodontally healthy groups (P > 0.05). TIMP‐1 concentration was higher in non‐smoker CP than smoker CP group (P < 0.05). MMP‐9/TIMP‐1 ratios were higher in smoker CP than non‐smoker CP group (P = 0.01). MMP‐8 concentrations, MMP‐8/TIMP‐1 and MMP‐9/TIMP‐1 ratios in CP group were not significantly different from those in periodontally healthy group (P > 0.05). Conclusions: Our findings of significantly elevated serum MMP‐9, MPO, NE together with decreased TIMP‐1 in smoker CP patients than non‐smokers support that smoking together with periodontal destruction may expose/predispose to cardiovascular diseases.     

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

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

Proteolytic Mediators in Gestational Diabetes Mellitus and Gingivitis

Background: This study evaluates levels of matrix metalloproteinase (MMP)‐8, MMP‐9, and tissue inhibitor of MP‐1 (TIMP‐1) in biofluids of women with gestational diabetes mellitus (GDM) and systemically healthy counterparts with different statuses of periodontal health. Methods: Seventy‐one women with GDM and gingivitis (Gg), 30 women with GDM and healthy periodontium (Gh), 28 systemically and periodontally healthy women (Hh), and 37 systemically healthy women with gingivitis (Hg) were evaluated. MMP‐8, MMP‐9, and TIMP‐1 levels were determined in gingival crevicular fluid (GCF), saliva, and serum by immunofluorometric and enzyme‐linked immunosorbent assays. Full‐mouth clinical periodontal parameters were recorded. Results: GCF and serum MMP‐8 concentrations, serum MMP‐9 concentrations, and serum MMP‐8/MMP‐1 and MMP‐9/MMP‐1 molar ratios were significantly higher in Gg compared with Hg group (P <0.05). Serum MMP‐8 levels and salivary TIMP‐1 levels were higher in Gh compared with Hg group (P <0.05) whereas salivary MMP‐8/TIMP‐1 molar ratio was lower in Gh compared with Hg group (P <0.05). Elevated concentrations of GCF MMP‐8 and MMP‐9 were found in Gg compared with Gh group (P <0.05). Significant correlations were found between local levels of biomarkers and clinical periodontal parameters in only GDM group. Conclusion: GDM may modulate both local and circulating levels of MMP‐8 especially when associated with gingivitis.     

The role of RANKL and MMP‐9 in the bone resorption caused by ameloblastoma

J Oral Pathol Med (2010) 39 : 592–598 Background: Ameloblastoma, a common odontogenic tumor located in jaws, generally leads to severe damage to patient’s complexion and masticatory function. To expand in jaws, ameloblastoma must have a mechanism of resorbing the surrounding bone. Our objective was to explore the bone‐resorption mechanism of ameloblastoma by observing the role of Receptor activator of nuclear factor kappa B ligand (RANKL) and matrix metalloproteinase‐9 (MMP‐9) in the bone‐resorption process. Methods: In the study, the expression of RANKL and MMP‐9 in ameloblastoma was detected using immunohistochemistry (IHC) and RT‐PCR. Then, co‐culture system of ameloblastoma cells and bone marrow cells from neonatal rabbit was erected to observe the potential of ameloblastoma cells to induce osteoclastogenesis. Finally, the induced osteoclasts were used for in vitro bone‐resorption assay. In the co‐culture system and the bone‐resorption assay, the selective inhibitor of RANKL and MMP‐9, osteoprotegerin (OPG) and tissue inhibitor of metalloproteinase‐1 (TIMP‐1) were, respectively, used for observing the role of RANKL and MMP‐9. Results: The expression of RANKL and MMP‐9 in ameloblastoma was confirmed. Ameloblastoma cells were found to induce bone marrow cells from neonatal rabbit differentiate into osteoclasts with bone‐resorption activity. In addition, OPG was found to, respectively, have markedly inhibitory effect on osteoclastogenesis (P < 0.01), and slightly inhibitory action on bone resorption (P < 0.05). Conclusions: Ameloblastoma cells had the potential to induce osteoclastogenesis. Moreover, RANKL played an essential role in the in vitro osteoclast formation and bone resorption induced by ameloblastoma cells.     

Cytokines,matrix metalloproteinases and tissue inhibitor of metalloproteinases‐1 in gingival crevicular fluid from patients with Papillon‐Lefèvre syndrome

The aim of the present study was to compare concentrations of cytokines, matrix metalloproteinases (MMPs) and a metalloproteinase inhibitor (TIMP‐1) in gingival crevicular fluids (GCF) from sites with gingival inflammation in 28 young patients with Papillon‐Lefèvre syndrome (PLS), and in age‐ and gender‐matched controls. Each group consisted of 17 females and 11 males with a mean age of 11.0 years (range 4–22 years). In both groups, anterior upper sites with a clinical diagnosis of gingival inflammation and with pockets ≤3?mm were selected for sampling of GCF, which was carried out with filter disks inserted into the gingival crevice until saturated. The concentrations of cytokines (IL‐1α, IL‐1β, TNF‐α, and IL‐8), matrix metalloproteinases (MMP‐1, MMP‐3, MMP‐8, and MMP‐9), and their tissue inhibitor (TIMP‐1) were analysed using commercial ELISA kits. Significantly higher levels of IL‐1β (P?P?P?P?P?相似文献   

Clinical and Biochemical Evaluation of Lozenges Containing Lactobacillus reuteri as an Adjunct to Non‐Surgical Periodontal Therapy in Chronic Periodontitis

Background: This study evaluates the effects on clinical and biochemical parameters of Lactobacillus reuteri–containing probiotic supplementation adjunctive to initial periodontal therapy in patients with chronic periodontitis (CP). Methods: Thirty patients with CP were included and divided into two groups. Every patient had, in each quadrant, ≥2 teeth each with approximal sites with a probing depth (PD) of 5 to 7 mm and gingival index (GI) of ≥2. The test group received scaling and root planing (SRP) and probiotic‐containing lozenges. The control group received SRP and placebo lozenges. Plaque index (PI), GI, bleeding on probing (BOP), PD, and attachment gain were measured. Gingival crevicular fluid (GCF) was sampled for the analysis of matrix metalloproteinase (MMP)‐8 and tissue inhibitor of metalloproteinase (TIMP)‐1 by enzyme‐linked immunosorbent assay. All evaluations were performed at baseline and on days 21, 90, 180, and 360. Results: Differences in intergroup comparisons of PI, GI, BOP, and PD were found to be significant (P <0.05) in favor of the test group at all time points. Decreased GCF MMP‐8 levels and increased TIMP‐1 levels were found to be significant up to day 180 (P <0.05). Mean values of attachment gain were significantly higher in the test group compared with the control group on days 90, 180, and 360. Conclusions: Lozenges containing L. reuteri may be a useful supplement in moderately deep pockets of patients with CP. Low MMP‐8 and high TIMP‐1 levels may indicate the role of the lozenges in reduction of inflammation‐associated markers up to day 180.