牙周非手术治疗对龈沟液MMP-8及其抑制剂TIMP-1水平的影响

目的: 比较慢性牙周炎患者非手术治疗前、后龈沟液MMP-8、TIMP-1水平的变化。方法: 选择慢性中重度牙周炎30例, 治疗前、后临床检查牙龈出血指数(SBI)、菌斑指数(PLI)、探诊深度(PD), 采集龈沟液, 采用双抗体夹心ABC-ELISA法检测MMP-8、TIMP-1的含量。采用SSPS 19.0软件包对数据进行配对样本t检验和多元相关分析。结果: 治疗后MMP-8、TIMP-1水平显著下降, MMP-8/TIMP-1 比率治疗前、后无显著差异。多元相关分析各变量间相关性, 发现MMP-8和龈沟液重量呈正相关关系。结论: 牙周非手术治疗能显著降低龈沟液MMP-8、TIMP-1水平, 从而减轻牙周组织损伤。     

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.     

Effects of phase I periodontal treatment on gingival crevicular fluid levels of matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase-1

OBJECTIVES: The aim of this study was to evaluate the effects of phase I periodontal treatment on gingival crevicular fluid (GCF) levels of matrix metalloproteinase (MMP)-3 and tissue inhibitors of metalloproteinase (TIMP)-1. METHODS: Plaque index, gingival index, pocket depth and clinical attachment loss were recorded and GCF samples were collected from 20 chronic periodontitis (CP) patients and 20 periodontally healthy controls (C) before treatment. CP patients received phase I periodontal treatment and all clinical parameters were recorded and GCF samples were collected once more after treatment. Assays were performed by an enzyme-linked immunosorbent assay. RESULTS: All of the clinical parameters improved significantly after the therapy (p<0.05). Baseline GCF levels of MMP-3 were significantly higher than C and that level was reduced significantly by treatment compared with baseline levels (p<0.05). Baseline GCF levels of TIMP-1 were lower than post-treatment levels and C (p<0.05). GCF levels of TIMP-1 increased significantly by treatment compared with baseline levels (p<0.05). CONCLUSION: This study shows that the clinical improvements after phase I periodontal therapy are accompanied by reduction in MMP-3 and increasing in TIMP-1 GCF levels.     

Effect of nonsurgical periodontal therapy on crevicular fluid levels of Cathepsin K in periodontitis

Objectives

Cathepsin K (CTSK), predominantly expressed in osteoclasts, is a potent extracellular matrix degrading enzyme that plays a critical role in osteoclast-mediated bone resorption. Its increased gingival crevicular fluid (GCF) levels in periodontal disease have been reported in a previous study. The present study has been carried out to assess the role of CTSK in periodontal disease and to determine the effect of periodontal treatment on CTSK concentration in GCF.

Design

60 subjects were divided into three groups (n = 20) based on gingival index (GI), probing pocket depth (PPD) and clinical attachment loss (CAL): healthy (group I), gingivitis (group II) and chronic periodontitis (group III). A fourth group (group IV) consisted of 20 subjects from group III, 6-8 weeks after nonsurgical periodontal therapy (scaling and root planing). GCF samples collected from each patient were quantified for CTSK using ELISA.

Results

The mean CTSK concentration in GCF was found to be the highest in group III, i.e. 55.55 pmol/l. The mean CTSK concentration in GCF in group I and group II was 5.95 pmol/l and 6.90 pmol/l respectively. The mean CTSK concentration in GCF in group IV decreased to 11.15 pmol/l, slightly more than that in groups I and II.

Conclusions

GCF CTSK levels increased in periodontitis and correlated negatively with clinical parameters like GI, PPD and CAL. CTSK levels decreased after nonsurgical treatment of periodontitis. Thus, CTSK can be considered as a ‘marker of osteoclastic activity’ in periodontal disease and also deserves further consideration as a therapeutic target.     

慢性牙周炎患者基础治疗前后龈沟液中瘦素水平变化的研究

目的：检测慢性牙周炎患者牙周基础治疗前后龈沟液中瘦素水平的变化。方法：选择轻、中、重度牙周炎患者共3组,每组11人,基础治疗前后收集龈沟液,采用酶联免疫法（Enzyme-Linked ImmunoSorbent Assay,ELISA）检测瘦素含量。结果：3组患者牙周基础治疗后龈沟液中瘦素水平均明显高于治疗前（P＜0.01）;瘦素与轻度牙周炎组治疗前的出血指数正相关（r=0.675）（P＜0.05）,与重度牙周炎组治疗前的探诊深度负相关（r=-0.799）（P＜0.01）;重度牙周炎组治疗后比治疗前的探诊深度减小（P＜0.01）,附着丧失减小（P＜0.05）。结论：龈沟液中瘦素含量变化与牙周炎的基础治疗密切相关,可以通过测定瘦素水平评估牙周炎治疗的临床疗效。     

Gingival crevicular fluid levels of monocyte chemoattractant protein-1 in periodontal health and disease

Objectives

Monocyte chemoattractant protein-1 (MCP-1) stimulates the chemotaxis of monocytes and also several cellular events associated with chemotaxis thus causes recruitment of inflammatory cells. Its increased gingival crevicular fluid (GCF) levels in periodontal disease have been reported in previous studies. The present study has been carried out to assess the role of MCP-1 in periodontal disease progression and also to determine the effect of periodontal treatment on MCP-1 concentration in GCF.

Design

A total of 60 subjects were divided into three groups (n = 20) based on gingival index (GI), probing pocket depth (PPD) and clinical attachment loss (CAL): healthy (group I), gingivitis (group II) and chronic periodontitis (group III). A fourth group (group IV) consisted of 20 subjects from group III, 6-8 weeks after treatment (i.e. scaling and root planing). GCF samples collected from each patient were quantified for MCP-1 using ELISA.

Results

The mean MCP-1 concentration in GCF was found to be the highest in group III, i.e. 72.60 pg/μl. The mean MCP-1 concentration in group I was 19.70 pg/μl and in group IV was 8.50 pg/μl. The mean MCP-1 concentration (37.00 pg/μl) in group II was found to lie in between the concentrations obtained in groups I and III.

Conclusions

GCF MCP-1 levels increased progressively with the progression of disease and decreased after treatment. Levels of MCP-1 correlated positively with clinical parameters like GI, PPD and CAL thus it can be considered as an inflammatory biomarker in periodontal disease and also deserves further consideration as a therapeutic target.     

Tissue inhibitor of metalloproteinases-1, matrix metalloproteinases-1 and -8, and collagenase activity levels in peri-implant crevicular fluid after implantation

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.     

SPR牙周基础治疗对慢性牙周炎患者龈沟液C反应蛋白的影响

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

Crevicular fluid endothelin-1 levels in periodontal health and disease

Background and Objective:  Endothelin-1 is a 21-amino-acid peptide with multifunctional regulation. Initial research indicated that endothelin-1 levels in the gingival crevicular fluid from patients with chronic periodontitis were higher than those in the gingival crevicular fluid from healthy subjects. The aim of the present study was to assess the relationship between the clinical parameters and the concentrations of endothelin-1 within the gingival crevicular fluid from inflamed gingiva and periodontitis sites and, subsequently, after the treatment of periodontitis sites.
Material and Methods:  A total of 60 subjects were divided into three groups – healthy (group I), gingivitis (group II) and chronic periodontitis (group III) – based on gingival index, pocket probing depth and clinical attachment loss. A fourth group consisted of 20 subjects from group III, 6–8 wk after treatment (i.e. scaling and root planing). Gingival crevicular fluid samples collected from each patient were quantified for endothelin-1 using an enzymatic immunometric assay.
Results:  Endothelin-1 was not detected in any sample from any of the study groups.
Conclusion:  The results showed that all the gingival crevicular fluid samples were negative for the endothelin-1 molecule. Therefore, endothelin-1 cannot be considered as a potential biomarker of periodontal disease progression.     

GCF and serum myeloperoxidase and matrix metalloproteinase-13 levels in renal transplant patients

Aim

The rationale of this study was to address whether local or systemic changes reflect proteolytic (matrix metalloproteinase-13) or oxidative (myeloperoxidase) stress in renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), in patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy.

Material and methods

Gingival crevicular fluid (GCF) samples were collected from sites with (GO+) and without GO (GO−) in CsA patients having GO; GO− sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum myeloperoxidase (MPO) and matrix metalloproteinase-13 (MMP-13) levels were determined by ELISA.

Results

GO+ sites in CsA patients having GO had elevated GCF MPO levels than those of CsA patients having no GO, tacrolimus and healthy subjects (p < 0.005), but comparable to those of gingivitis. GCF MPO levels were higher in GO+ compared to GO− sites in CsA patients having GO (p < 0.05). Patient groups had similar, but higher GCF MMP-13 levels than healthy group.

Conclusions

These results show that CsA and tacrolimus therapy have not a significant effect on GCF MPO and MMP-13 levels, and gingival inflammation seems to be the main reason for their elevations.     

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.     

慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、MMP-8/TIMP-1水平的关系

目的： 探讨慢性牙周炎患者唾液中miR-146a的表达及其与龈沟炎症、基质金属蛋白酶8（MMP-8）、基质金属蛋白酶抑制剂1（TIMP-1）水平的关系。方法： 选择2015年3月—2017年1月间收治的慢性牙周炎患者68例作为慢性牙周炎组,同期在本院进行体检的健康志愿者50例作为正常对照组。检测2组研究对象唾液中miR-146a的表达量,龈沟液中炎症因子、MMP-8/TIMP-1的水平及牙周临床症状指标。采用SPSS24.软件中的Pearson检验评估慢性牙周炎患者唾液中miR-146a的表达量与病情严重程度的相关关系。结果： 慢性牙周炎患者唾液中miR-146a的表达量显著高于正常对照组（P<0.05）,龈沟液中炎症因子（IL-1β、IL-6、IL-35、TNF-α）水平、牙周临床症状指标（PD、AL、PLI、BI）以及龈沟液中MMP-8、TIMP-1的水平显著高于正常对照组（P<0.05）。Pearson检验发现,慢性牙周炎患者唾液中miR-146a表达量与龈沟炎症程度、牙周临床症状严重程度及MMP-8/TIMP-1水平呈正相关。结论： 慢性牙周炎患者唾液中miR-146a表达量异常增高,且与龈沟炎症程度、牙周损伤程度一致。     

Protease inhibitor levels in periodontal health and disease

Kretschmar S, Yin L, Roberts F, London R, Flemmig TT, Arushanov D, Kaiyala K, Chung WO. Protease inhibitor levels in periodontal health and disease. J Periodont Res 2012; 47: 228–235. © 2011 John Wiley & Sons A/S Background and Objective: Our previous study showed that protease inhibitors were attenuated by the periodontal pathogen Porphyromonas gingivalis in cultured gingival epithelial cells. We hypothesize that fewer protease inhibitors would be present in more advanced periodontal disease sites, where the level of P. gingivalis may be high. The goal of this study was to investigate the relationship between the protease inhibitor [secretory leukocyte protease inhibitor (SLPI), elastase‐specific inhibitor (ELAFIN) and squamous cell carcinoma antigen (SCCA)] levels in gingival crevicular fluid and the number of P. gingivalis micro‐organisms in subgingival plaque. Material and Methods: Plaque samples from subjects without (n = 18) and with moderate to advanced periodontitis (n = 41) were used to quantify P. gingivalis using real‐time PCR. Protease inhibitor levels in the gingival crevicular fluid of all the subjects were determined by ELISA. Results: P. gingivalis was detected in 68.3% of patients with periodontitis, while 16.7% of subjects without periodontitis had a detectable level of P. gingivalis. Patients with periodontitis and P. gingivalis in their plaque exhibited lower SLPI and ELAFIN levels (p < 0.001) compared with control subjects without periodontitis. Secretory leukocyte protease inhibitor was also reduced (p < 0.05) in gingival crevicular fluid of periodontitis patients without a detectable level of P. gingivalis. Periodontitis patients with high vs. low levels of P. gingivalis exhibited reciprocal mean levels of SLPI and ELAFIN concentrations. Conclusion: The reduced concentrations of SLPI and ELAFIN may contribute to the loss of host protective capacity and increase susceptibility to breakdown from chronic infection. The work of this investigation may aid in finding diagnostic and prognostic markers in periodontal health and disease and may also help in finding pharmacological targets directed against periodontal inflammation.     

慢性牙周炎龈沟液中硫离子水平与临床相关性研究

目的:分析慢性牙周炎(CP)患者龈沟液中硫离子(su lfides)水平的变化与临床牙周指数的相关关系及其对诊断预后的意义。方法:采用金刚牙周诊断仪进行龈沟液硫离子和牙周临床指标测定。选定实验组(T):36例慢性牙周炎患者,57颗牙位,共342个位点。其中健康牙位(T1)21颗,位点126个;炎症牙位(T2)36颗,位点216。对照组(C):全身及牙周健康者8例,16颗牙位,共96个位点。测定所选位点龈沟液(GCF)中硫化物水平(su lcussu lph ide level,SUL),牙周袋探诊深度(prob ing depth,PD),牙周临床附着丧失水平(c lin ical attachm ent level,CAL),龈沟出血指数(su lcus b leed ing index,SB I)。所有统计结果均采用SPSS11.0进行统计学分析。结果:1)牙周健康对照组(C)GCF中硫离子SUL的浓度均值为(0.0648±0.0169)pg/mL,明显低于慢性牙周炎炎症牙位组(T2)(0.3249±0.0489)pg/mL及慢性牙周炎健康牙位组(T1)(0.1160±0.0271)pg/mL;慢性牙周炎炎症牙位组(T2)GCF中硫离子(SUL)的浓度均值均高于正常对照组及慢性牙周炎健康牙位组(T1)。2)经相关性分析,慢性牙周炎炎症牙位组(T1)GCF中SUL的浓度均值与PD、SB I和CAL均呈正相关关系。而慢性牙周炎健康牙位组(T1)及正常对照组(C)GCF中SUL的浓度均值与PD、SB I及CAL间无相关性。结论:慢性牙周炎(CP)炎症牙位组龈沟液中硫离子(SUL)的浓度均值与牙周临床指标之间具有相关关系,其水平的高低变化可客观反映牙周组织的炎症状态。     

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目的 探讨牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)W83、ATCC33277刺激人牙周膜成纤维细胞(HPDLFs)分泌基质金属蛋白酶-1(MMP-1)、金属蛋白酶组织抑制剂-1(TIMP-1)的变化.方法 本研究于2010年9月至2011年6月在中国医科大学口腔医学院中心实验室进行.将P.gingivalis W83、ATCC33277作用于HPDLFs0、6、12、24、48h后,运用酶联免疫吸附法(ELISA)检测细胞上清液中MMP-1、TIMP-1质量浓度变化,并计算MMP-1/TIMP-1值.结果 P.gingivalis感染HPDLFs的MMP-1和TIMP-1表达均增强,并呈时间依赖性;且MMP-1/TIMP-1值明显高于未感染的对照组(P＜0.05).P.gingivalis W83感染后MMP-1/TIMP-1值明显高于P.gingivalis ATCC33277(P＜ 0.05).结论 P.gingivalis具有促进HPDLFs分泌MMP-1、TIMP-1的作用,且可造成牙周组织破坏;P.gingivalis W83降解细胞外基质的能力高于P.gingivalisATCC33277.     

正常人、牙龈炎和牙周炎患者龈沟液内一氧化氮含量的检测

目的：检测正常人和牙周病患者龈沟液中NO含量,探讨NO在牙周病发病过程中的作用。方法：选择牙周健康组20例,牙龈炎组22例,慢性牙周炎组32例,分别采集龈沟液标本,免疫荧光法检测龈沟液内NO的含量。结果：慢性牙周炎患者和牙龈炎患者龈沟液内NO含量与牙周健康组相比均有高度显著性差异（P〈0．01）,慢性牙周炎患者龈沟液内NO含量与牙龈炎组相比有高度显著性差异（P〈0．01）。结论：牙周健康者、牙龈炎患者、慢性牙周炎患者龈沟液中能检测出NO的存在,NO参与了慢性牙周炎的发展过程,龈沟液内NO含量与慢性牙周炎炎症程度密切相关。     

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

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

Gingival crevicular fluid lactoferrin levels in adult per iodontitis patients

The present study was designed to determine in a cross-sectional study whether there was any relationship between the levels of lactoferrin in gingival crevicular fluid and clinical periodontal parameters. Crevicular fluid was collected from individual sites using standardized filter paper strips (clinically healthy sites, N=23; periodontitis sites, n=66) and evaluated for lactoferrin by enzyme-linked immunosorbent assay. The data showed that: (1) the total amounts of lactoferrin were 0.003-0.021 ng (30 second sample) (average 0.009±0.005 ng) in a clinically healthy periodontium group and 0.016-3.847 ng (30 second sample) (average 0.575±0.069 ng) in adult periodontitis patients (statistically significantly higher in adult periodontitis patients); and (2) the total amounts of lactoferrin were significantly correlated with clinical parameters,