慢性牙周炎基础治疗前后龈沟液中MMP-2,TIMP-2水平的变化

目的:观察慢性牙周炎患者在牙周基础治疗前后龈沟液(gingivalcrevicularfluid,GCF)中基质金属蛋白酶2(matrixmetalloprotein-ase -2,MMP -2)和基质金属蛋白酶抑制剂2(tissueinhibitorsofmatrixmetalloprotein ase -2,TIMP- 2)水平的变化。方法:采用滤纸条袋内取样法取42例患者42个牙位治疗前后的龈沟液(GCF)样本,同时取42例健康对照的42个牙位的GCF样本,用酶联吸附免疫法(enzyme linkedimmunosorbentas say, ELISA)检测其中的MMP- 2,TIMP- li2水平。结果:牙周基础治疗后MMP- 2,TIMP -2的总量均较治疗前显著下降(P<0. 01 ),治疗前后MMP -2,TIMP -2的总量均与对照组有显著差别(P<0. 01 ),治疗后MMP- 2与TIMP -2的比值较治疗前显著下降,同时高于对照组MMP -2与TIMP -2的比值。MMP -2的含量与TIM-P 2的含量呈显著正相关(P<0. 01)。结论:成功的牙周基础治疗后,MMP -2的总量相对明显下降, TIMP- 2的总量相对升高。     

大鼠正畸牙移动前后龈沟液量变化的研究

目的:研究大鼠正畸牙移动前后龈沟液(GCF)量的变化.方法:选择6周龄,健康雌性SPF级Wistar大鼠20只,实验侧安装正畸加力装置,对照侧结扎丝环扎第一磨牙,不施加任何矫治力.分别测量实验前1周及实验后1-7周实验侧与对照侧上颌第一磨牙龈沟液的量,使用SPSS 11.0统计软件包,对GCF重量进行统计学分析,比较安装加力装置前后实验侧与对照侧龈沟液量的变化.结果:无论实验侧还是对照侧,安装加力装置后的GCF量均高于安装前(P＜0.05),安装加力装置后各周之间的GCF量差别无统计学意义(P＞0.05);安装加力装置后各周,实验侧的GCF量均高于对照侧(P＜0.05).结论:大鼠牙齿移动后龈沟液的量有明显增加.     

大鼠舌鳞癌诱变过程中MMP-3、TIMP-1的表达变化及相关性研究

目的:研究基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶组织抑制因子-1(TIMP-1)在大鼠舌鳞癌诱变过程中转录水平表达变化及两者之间的相关性。方法:60只SD大鼠随机分为3组,每组20只,正常对照组,给予正常饮食;另2组每天喂养0.002%4-硝基喹啉-1-氧化物(4NQO),于16周和24周处死;16周组可见上皮异常增生,而24周组已经为舌鳞癌,分别选取8个标本作检测。各组利用定量逆转录聚合酶链反应(qRT-PCR)检测MMP-3、TIMP-1在不同病变时期舌组织中转录水平的表达变化。结果:MMP-3及TIMP-1的mRNA于正常对照组相对表达量较少,上皮异常增生组相对表达量较正常组显著增加,舌鳞癌组的相对表达量最高。MMP-3与TIMP-1的mRNA表达之间成显著相关性。结论:MMP-3与TIMP-1随癌变的发生表达显著上调,两者间平衡失调可导致舌鳞癌的发生及发展。     

正畸应力变化下牙龈组织α-SMA、Ⅰ型和Ⅲ型胶原变化及龈沟液MMP-2、TIMP-2的表达

目的: 探讨正畸应力变化下牙龈组织α平滑肌肌动蛋白（α-SMA）、Ⅰ型及Ⅲ型胶原变化以及龈沟液基质金属蛋白酶2（MMP-2）、金属蛋白酶组织抑制因子2（TIMP-2）的表达。方法: 选取南昌大学附属口腔医院2018年4月—2019年4月收治的正畸患者74例,随机分为A组（24例,0 g力）、B组（25例,75 g力）、C组（25例,150 g力）3组。于加力0、4周采集患者部分牙龈组织,采用免疫组织化学染色法检测α-SMA、Ⅰ型及Ⅲ型胶原表达水平。在加力后0、2、4周收集龈沟液,采用酶联免疫吸附法检测MMP-2、TIMP-2表达水平。采用Spearman相关性分析不同正畸应力与α-SMA、Ⅰ型胶原、Ⅲ型胶原、MMP-2、TIMP-2表达水平的相关性。采用SPSS 19.0软件包对数据进行统计分析。结果: 加力2周及加力4周,3组患者龈沟液MMP-2、TIMP-2表达水平依次升高（P<0.05）;加力4周后,3组患者牙龈组织中α-SMA、Ⅰ型及Ⅲ型胶原水平依次升高（P<0.05）;不同正畸应力与MMP-2、TIMP-2、α-SMA、Ⅰ型胶原及Ⅲ型胶原表达水平均呈正相关（P<0.05）。结论: 龈沟液TIMP-2、MMP-2表达水平以及肌成纤维细胞表达与正畸应力变化有关,可能在正畸治疗的牙周组织改建中发挥重要作用。     

人类牙周膜组织中MMP-2及TIMP-2的比较研究

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。     

糖基化终产物对人牙周膜细胞MMP-3及TIMP-1表达的影响

目的:探讨糖基化终产物(AGEs)对人牙周膜细胞MMP-3及TIMP-1蛋白表达的影响,进一步阐明糖尿病牙周病发病过程中细胞外基质代谢调节的分子机制.方法:将糖基化终产物修饰的人血清白蛋白与人牙周膜细胞在体外共同培养,用免疫荧光法及ELISA技术检测人牙周膜细胞MMP-3、TIMP-1蛋白表达.结果:人牙周膜细胞在体外表达MMP-3,培养3d后,MMP-3表达明显增强,与对照组相比具有显著性差异.而TIMP-1表达减弱,与对照组相比不具有显著性差异.结论:糖基化终产物可显著影响人牙周膜细胞MMP-3的表达,为AGEs参与糖尿病牙周病发病机制提供了直接证据.     

周期性牵张力对人牙周膜细胞MMP-3及TIMP-1表达的影响

目的 :探讨周期性牵张力对人牙周膜细胞MMP -3及TIMP -1蛋白表达的影响 ,进一步阐明正畸牙牙周组织改建过程中ECM代谢调节的分子机制 .方法 :通过体外细胞培养加载系统 ,选择频率为 6周 /min ,弹性基底膜发生 12 %形变率的周期性牵张力 ,利用免疫组化及夹心ELISA检测技术 ,观察HPDLC表达MMP -3及TIMP -1的相对强度。结果 :HPDLC在体外表达MMP -3及TIMP -1。加载 3d后 ,MMP -3表达明显增强 ,与对照组相比具有显著性差异。而TIMP -1对机械力无应答。结论 :在一定条件下 ,周期性牵张力可显著影响HPDLCMMP -3蛋白的表达。为机械力作用下MMP -3参与牙周组织ECM代谢提供了直接证据     

不同年龄组大鼠正畸力作用下龈沟液中基质金属蛋白酶的表达

目的:观察2个年龄组大鼠牙受正畸力后龈沟液中基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)含量的变化.方法:选择6周龄和12周龄雌性Wistar大鼠各20只,采用粘结剂联合磨沟周位法在上颌安装正畸加力装置,牵引一侧上颌第一磨牙向近中,另一侧作为对照侧.每周提取大鼠实验牙及对照牙的龈沟液,应用SDS-PAGE、Western印迹和Gel-Doc凝胶成像系统定量检测龈沟液中MMP-1与MMP-9含量的改变.采用SPSS11.0软件包对实验各周龈沟液中MMP-1和MMP-9的含量进行方差分析.结果:6周龄组大鼠和12周龄组大鼠龈沟液中MMP-1及MMP-9含量的基础水平以及在加力第1周时的水平无统计学差异(P＞0.05).但12周龄组比6周龄组稍高.加力早期,12周龄组大鼠MMPs含量增长速度快于6周龄组;后期,12周龄组大鼠较6周龄组大鼠的MMPs含量下降速度慢.结论:12周龄大鼠受正畸力后,龈沟液中MMPs的表达较6周龄组高,12周龄大鼠较6周龄大鼠的正畸牙根吸收可能更严重.     

MMP-3及TIMP-1在正畸牙周组织改建过程中的表达

目的：探讨大鼠正畸牙周组织改建过程中基质金属蛋白酶－３（ＭＭＰ－３）和金属蛋白酶组织抑制因子－１（ＴＩＭＰ－１）与正畸牙齿移动的关系。方法：在ＳＤ成年大鼠上颌左侧第一磨牙上颌切牙之间安置正畸装置,建立大鼠上移动实验模型。于牙齿移动１、３、５、７、１４ｄ后取材分别进行免疫组化染色,图像分析,观察ＭＭＰ－３和ＴＩＭＰ－１表达的变化。结果：牙齿移动１ｄ后,牙周组织细胞ＭＭＰ－３表达增强,５ｄ后ＭＭＰ－３     

正畸牙齿移动过程中龈沟液前列腺素E_2的变化

多种生物活性物质在正畸牙齿移动过程中起作用，其中前列腺素Ｅ２（ＰＧＥ２）一直受到关注。本研究选取１３例拔除双侧上颌第一双尖牙的正畸患者，用ＰａｕｌＧｊｅｓｓｉｎｇ设计的上颌尖牙后移簧（ＰＧＣＲＳ）远中移动尖牙，在严格控制口腔卫生的条件下，以滤纸条法取受力前、受力后１小时、２４小时及１６８小时一侧尖牙远中的龈沟液（ＧＣＦ）。用放射免疫法（ＲＩＡ）测ＧＣＦ—ＰＧＥ２的含量。结果显示受力前即可检出ＧＣＦ－ＰＧＥ２，平均为４６．７３９ｐｇ／ｍｌ。受力后１小时、２４小时及１６８小时分别为１２６．６０９、２０８．８７８和１２１．７２８ｐｇ／ｍｌ。均较受力前显著增加（Ｐ＜０．０５）。表明牙受力初期ＧＣＦ－ＰＧＥ２含量增加，且在２４小时达到峰值，提示ＧＣＦ－ＰＧＥ２含量增加与牙齿的受力移动有关。ＧＣＦ－ＰＧＥ２提供了一种研究人牙齿移动过程中牙周组织生化指标变化的活体检测方法     

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.     

种植体周龈沟液中EMMPRIN及其配体CyPA的表达

［摘要］ 目的 判断种植体周龈沟液中细胞外基质金属蛋白酶诱导因子(extracellular matrix metalloproteinase inducer,EMMPRIN)及亲环素A(cyclophilin A,CyPA)的表达情况。方法 收集种植体行使功能1年以上的患者(种植组)的种植体周龈沟液以及无缺牙、无牙周炎患者(对照组)的天然牙龈沟液,并进行相关临床检查,ELISA法检测龈沟液中EMMPRIN及CyPA的表达,应用SPSS 17.0软件包进行统计学分析,分析两者的表达与各临床检查指标之间的关系,比较两组间的差异。结果 种植组中,种植体周龈沟液(peri-implant crevicular fluid,PICF)总量与EMMPRIN、CyPA总量显著正相关;EMMPRIN总量与PICF、CyPA总量、牙龈指数(gingival index,GI)及改良菌斑指数(modified plaque index,mPLI)成显著正相关;CyPA总量与PICF、EMMPRIN总量及GI成显著正相关。在种植组与对照组之间,PICF/GCF、EMMPRIN、CyPA三者总量均无显著差异。结论 EMMPRIN及其配体CyPA在种植体周龈沟液中的表达与种植体周龈沟液的分泌量呈显著正相关。     

The effects of orthodontic tooth movement on the glycosaminoglycan components of gingival crevicular fluid

Abstract In this study, gingival crevicular fluid (GCF) was collected from around a canine tooth, in children, before and during orthodontic tooth movement. The aim was to identify and quantify the glycosaminoglycan (GAG) components of GCF and relate them to tooth movement, gingival inflammation, plaque accumulation, pocket probing depth and GCF volume recorded at the site of sampling. GAG in GCF samples, collected for a 15-min period into microcapillary tubes, were separated electrophoretically, stained with Alcian blue and quantified using a laser densitometer. 2 GAG components of hyaluronic acid (HA) and chondroitin sulphate (CS) were identified. The increase in GCF volume during orthodontic tooth movement was only partly due to increased gingival inflammation, GAG levels varied with different types of orthodontic tooth movement. In GCF, levels of CS, in particular, may reflect the changes in the deeper periodontal tissues which could be monitored during orthodontic tooth movements.     

三种冠边缘设计对龈沟液中内毒素含量的影响

目的:通过测定冠修复前后龈沟液内毒素含量的变化,评价3种冠边缘设计对牙龈状况与龈沟液中内毒素含量的影响。方法:将要求烤瓷熔附金属全冠修复的就诊患者,随机分为A:肩台组、B:浅凹型肩台组、C:刃状边缘组,每组各10例。分别将修复前、修复后1个月、6个月的龈沟液样本内毒素量、龈沟液量及牙龈指数进行比较,统计学处理。结果:3组患牙冠修复后1个月各项指标明显上升。冠修复后6个月A、B两组各项指标下降至接近牙体预备前水平;C组内毒素水平无下降,牙龈指数、龈沟液量与修复后1月相比有所下降但无统计学意义。结论:只要按要求制作,A、B组龈边缘形式的修复均具有良好的效果,修复效果明显优于C组。     

Levels of tissue inhibitor of metalloproteinases-1 and matrix metalloproteinases-1 and -8 in gingival crevicular fluid following treatment with enamel matrix derivative (EMDOGAIN)

The mechanism of enamel matrix derivative (EM D) action on the periodontal wound healing process is not well understood. However, earlier in vitro studies from our laboratory demonstrated that EMD stimulated the proliferation of both periodontal ligament and gingival fibroblast cells. Therefore, the purpose of this study was to further evaluate the effect of EMD on the early wound healing process by assessing the protein levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in gingival crevicular fluid (GCF). Sixteen patients, each of whom had one or two pairs of infrabony defects located contralaterally in the same arch, were included in this clinical trial. Thirty-six infrabony defects were randomly assigned treatment with flap surgery plus EMD or flap surgery plus placebo. At baseline and at 2, 4 and 12 week follow-up evaluation visits, GCF was sampled with paper strips. After determination of GCF volume, TIMP-1, MMP-1 and MMP-8 GCF levels were measured by an enzyme-linked immunosorbent assay. Intragroup analysis: At week 2 following surgery, when compared to baseline all parameters in each study group, except MMP-1, significantly increased (p<0.05). There were no significant differences between 4 or 12 weeks and baseline in either study group. Intergroup analysis: At 4 weeks after surgery, GCF volume and TIMP-1 levels showed a significant decrease (p<0.05) in the EMD group, when compared to the placebo group. MMP-1 levels at weeks 2, 4 and 12, and MMP-8 levels at weeks 4 and 12 were significantly lower (p < 0.05) in the EMD group compared to the placebo group. EMD compared to placebo treated sites demonstrated a more rapid return to baseline levels of TIMP-1, MMP-1 and MMP-8. These findings suggest that treatment with flap surgery and EMD, compared to flap surgery with placebo, accelerated healing at an earlier stage of wound healing following surgery.     

Collagenase-2 (MMP-8) and collagenase-3 (MMP-13) in adult periodontitis: molecular forms and levels in gingival crevicular fluid and immunolocalisation in gingival tissue

AIM: To determine the cellular and molecular forms of MMP-8 (collagenase-2) and MMP-13 (collagenase-3) associated with chronic adult periodontitis by examining the species present in gingival crevicular fluid (GCF) and enzyme distribution in gingival tissue. METHODS: 30-s GCF samples were collected directly from the periodontal pockets of 12 untreated patients using filter paper strips. After elution into buffer, the samples were examined by Western immunoblotting with polyclonal antibodies for MMP-8 and MMP-13 and quantification by scanning image analysis. Individual band intensities were expressed as a percentage of total sample absorbance and mean patient values were calculated. Gingival tissue from 6 patients was fixed in formalin and embedded in paraffin wax. MMP-8 and MMP-13 were localised using the same antibodies and an avidin-biotin-peroxidase detecting system. Double staining was performed with a contrasting substrate reaction. RESULTS: The majority of MMP-8 staining in pre-treatment GCF was present in 80, 75 and 60 kD bands corresponding to prepro-, pro- and active forms of PMN-type enzyme. 43 and 38 kD bands evidently represented active, fibroblast-type MMP-8. Immunoreactivities at >100 kD and < or =30 kD were probably enzyme-inhibitor complex and degraded fragments, respectively. MMP-13 was seen mainly as 60 kD proenzyme with some 40 kD active enzyme and a small proportion of >100 kD complex. The percentages of MMP-8 PMN-type enzyme and MMP-13 proenzyme bands correlated significantly with gingival and bleeding indices (p<0.05). Immunohistochemistry demonstrated MMP-8 in PMNs, sulcular epithelial and also plasma cells in inflamed gingival connective tissue. MMP-13 immunoreactivity was detected in the sulcular epithelium and in macrophage-like cells. CONCLUSION: Multiple species and elevated levels of both MMP-8 and MMP-13 from many rather than single cellular sources in the diseased periodontium are identified in untreated periodontitis GCF and active forms contribute to GCF collagenase activity.     

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目的对脱位牙再植术后龈沟液（gingival cervicular fluid,GCF）内基质金属蛋白酶（matrix metalloproteinase,MMP）-2和基质金属蛋白酶组织抑制因子（tissue inhibitor of metalloproteinase,TIMP）-2的含量进行定量分析,并研究术后不同时间段其变化规律,探讨GCF内MMP-2及TIMP-2在牙周组织改建中的作用。方法选择2010年8月至2012年3月在温州医科大学附属口腔医院就诊的20例7~13岁上颌中切牙外伤脱位患儿,参照国际牙外伤协会的年轻恒牙外伤管理指南2（2007年版）中的处理步骤行脱位牙再植术并进行固定,分别于术后第1、3、7天采集再植牙的GCF并定量测定其中MMP-2及TIMP-2的含量,比较不同时间段GCF内MMP-2及TIMP-2含量的差别,并比较MMP-2与TIMP-2比值（MMP-2/TIMP-2）的差别。结果术后第1天GCF内MMP-2及TIMP-2的含量均显著高于第3天和第7天,且术后第1天两者比值亦显著高于第3和第7天。结论脱位牙再植术后前期阶段（1~3 d）GCF中MMP-2及TIMP-2的含量变化剧烈,后期阶段（4~7 d）变化程度较小。提示MMP-2及TIMP-2可能在牙再植后牙周组织改建过程中扮演重要角色。     

龈沟液中性多形核白细胞数目的重复性研究

目的为研究一些溶酶体源的酶成分与PMN的关系提供一个可靠的取样方法。方法采用龈沟灌洗的方法，间隔24h重复取GCF样本，2～5倍稀释后于光学显微镜下计数PMN，观察两次的GCF中PMN数目的变化。结果两次的PMN数；数没有统计学差别且强正相关（r＝0．988）。结论在临床指标较为一致的情况下，可用第2天的PMN数目反映第1天的状况。