Herrmann J M, Gonzáles J R, Boedeker R H, Vonholdt J, Meyle J
Department of Periodontology, Dental School, University of Giessen, Germany.
J Clin Periodontol. 2001 Jan;28(1):31-7. doi: 10.1034/j.1600-051x.2001.280105.x.
BACKGROUND, AIMS: A new microassay for the detection of elastase activity (EA) in gingival crevicular fluid (GCF) has been established. GCF was collected with Periopaper strips and quantified in a Periotron.
Enzyme activity was measured in a microtiter plate reader, using a fluorometric assay. To ensure quality and precision of the assay, recovery rates were determined at different activities with a recovery of >90%. In a 2nd step, stability of the enzyme was investigated during storage at room temperature, +4 degrees C, -22 degrees C, -88 degrees C. GCF samples retained elastase activity of almost 100% after a storage of 3 days at -22 degrees C. In a group of 12 healthy volunteers, elastase activity was assayed throughout an 18 day experimental gingivitis protocol.
Median activity increased from 481 microU/microl at baseline to 1444 microU/microl at day 18, which was accompanied by the development of the signs of gingivitis. The increase of EA during the experimental phase of the study was highly significant (p<0.001) and correlated well with the increasing severity of gingivitis.
The data suggest that elastase activity in GCF is an excellent quantitative measure of gingival inflammation.
背景、目的：已建立一种用于检测龈沟液（GCF）中弹性蛋白酶活性（EA）的新微量测定法。使用Periopaper试纸条收集GCF，并在Periotron中进行定量。
采用荧光测定法在酶标仪中测量酶活性。为确保测定的质量和精密度，在不同活性水平下测定回收率，回收率>90%。第二步，研究酶在室温、+4℃、-22℃、-88℃储存期间的稳定性。GCF样本在-22℃储存3天后弹性蛋白酶活性保留率近100%。在一组12名健康志愿者中，在为期18天的实验性牙龈炎方案中全程测定弹性蛋白酶活性。
中位数活性从基线时的481微单位/微升增加至第18天的1444微单位/微升，同时伴有牙龈炎体征的出现。研究实验阶段EA的增加非常显著（p<0.001），且与牙龈炎严重程度的增加密切相关。
数据表明，GCF中的弹性蛋白酶活性是牙龈炎症的一种优秀定量指标。
