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Detection of gingival crevicular fluid MMP-8 levels with different laboratory and chair-side methods.

机译:用不同的实验室和椅旁方法检测龈沟液MMP-8水平。

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OBJECTIVE: The aim of the study was to compare four methods for gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 detection. METHODS: Matrix metalloproteinase-8 levels from 20 GCF samples from two periodontally healthy subjects, 18 samples from two patients with gingivitis and 45 samples from six patients with moderate to severe periodontitis, altogether 83 samples, were analysed using (1) a time-resolved immunofluorometric assay (IFMA), (2) an MMP-8 specific chair-side dip-stick test, (3) a dentoAnalyzer device and (4) the Amersham ELISA kit. Western immunoblot using same monoclonal anti-MMP-8 as in IFMA and dentoAnalyzer was used to identify molecular forms of MMP-8 in GCFs. RESULTS: Correlation between IFMA and dentoAnalyzer results calculated with Spearman's correlation coefficient was 0.95 (P = 0.01). The chair-side dip-stick test results were well in line with these assays. Periodontitis sites with unstable characteristics were differentiated with these methods. The Amersham ELISA results were not in line with the findings by other methods. CONCLUSIONS: Immunofluorometric assay and dentoAnalyzer can detect MMP-8 from GCF samples and these methods are comparable. Using Western immunoblot, it was confirmed that IFMA and dentoAnalyzer can detect activated 55 kDa MMP-8 species especially in periodontitis-affected GCF. dentoAnalyzer is among the first quantitative MMP-8 chair-side testing devices in periodontal and peri-implant diagnostics and research.
机译:目的:本研究的目的是比较四种检测牙龈沟液(GCF)基质金属蛋白酶(MMP)-8的方法。方法:使用(1)时间分辨分析了来自两个牙周健康受试者的20个GCF样品,两个牙龈炎患者的18个样品和六个中度至重度牙周炎患者的45个样品,总共83个样品的基质金属蛋白酶-8水平免疫荧光测定(IFMA),(2)MMP-8特定的椅子侧浸量尺测试,(3)dentoAnalyzer设备和(4)Amersham ELISA试剂盒。使用与IFMA和dentoAnalyzer相同的单克隆抗MMP-8抗体进行Western免疫印迹,以鉴定GCF中MMP-8的分子形式。结果:IFMA与使用Spearman相关系数计算的dentoAnalyzer结果之间的相关性为0.95(P = 0.01)。椅子旁的浸油尺测试结果与这些测定非常吻合。用这些方法区分出特征不稳定的牙周炎部位。 Amersham ELISA结果与其他方法的发现不符。结论:免疫荧光测定法和dentoAnalyzer可以检测GCF样品中的MMP-8,这些方法具有可比性。使用Western免疫印迹证实,IFMA和dentoAnalyzer可以检测到激活的55 kDa MMP-8物种,尤其是在牙周炎影响的GCF中。 dentoAnalyzer是用于牙周和种植体周围诊断和研究的首批定量MMP-8椅旁测试设备之一。

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