Nomura T, Ishii A, Oishi Y, Kohma H, Hara K
Department of Periodontology, Niigata University School of Dentistry, Japan.
Oral Dis. 1998 Dec;4(4):231-40. doi: 10.1111/j.1601-0825.1998.tb00286.x.
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.
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.
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.
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.
对从健康受试者、牙龈炎患者和牙周炎患者获取的龈沟液(GCF)中金属蛋白酶组织抑制剂(TIMPs)水平、胶原酶活性以及基质金属蛋白酶(MMP)-1和-8的免疫反应性进行全面评估,并通过主成分分析来分析牙周疾病中牙周组织破坏与GCF成分之间的关系。
用无菌纸条从10名牙龈炎患者和11名牙周炎患者中采集GCF。10名志愿者作为临床健康对照。通过酶联免疫吸附测定法(ELISA)测量GCF中TIMP-1和-2的蛋白量,并在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)后使用图像分析通过功能测定法确定活性和经氨基苯汞乙酸(APMA)激活的胶原酶活性。
患病组GCF中TIMP-1水平以及活性和潜在胶原酶活性均显著高于健康组。仅在所有受试者的29%中可检测到TIMP-2(平均值:2.06纳克)。蛋白质印迹分析表明,MMP-8是患病组GCF中的主要间质胶原酶。使用临床参数和GCF成分进行的主成分分析表明,在评估其测量值与牙周疾病的相关性时,成分一至三占总变异的87%。
我们对牙周病患者GCF中的MMPs和TIMPs进行了功能和免疫学特征分析。主成分分析表明成分一至三解释了总变异的87%,并进一步表明较高的胶原酶活性(尤其是活性胶原酶)将是评估牙周炎发病机制的重要标志物。因此这些观察结果可能具有重要的治疗和诊断意义。