Eley B M, Cox S W
Department of Periodontology, Dental School, King's College School of Medicine and Dentistry, Denmark Hill, London.
Br Dent J. 1995 Feb 25;178(4):133-9. doi: 10.1038/sj.bdj.4808681.
Our recent work has developed specific assays for bacterial dipeptidyl peptidase (DPP) and trypsin-like proteases and we have found them in gingival crevicular fluid (GCF). The purpose of this study was to determine whether their levels reduce following periodontal treatment of chronic periodontitis patients. The probing depth, probing attachment level, gingival index, gingival bleeding index and plaque index were measured at mesio-buccal sites of molars and premolars in 25 untreated patients. At a second visit GCF was collected on filter paper strips for 30 seconds. GCF volumes were determined with a Periotron and the samples eluted into buffer. The patients than received oral hygiene instruction, supra- and subgingival scaling and other appropriate non-surgical treatment. Four weeks later GCF was collected from the same 16 sites and the clinical parameters were measured again. DPP-like activity was determined fluorometrically with Ala-Pro-AFC at pH 8.0 with and without heating to 60 degrees C for 30 minutes. The heat sensitive portion was taken as bacterial DPP activity. Bacterial trypsin-like protease activity was assayed with Z-Val-Lys-Lys-Arg-AFC at pH 7.0 with 2 mM dithiothreitol and 10 microM Z-Phe-Ala-CH2. Following treatment there were marked reductions in clinical parameters, enzyme total activities and concentrations. All reductions were statistically significant at patient and site level using either individual patient or pooled patient data. Bacterial proteases appear to reflect the clinical status and may be of value in monitoring chronic inflammatory periodontal disease.
我们最近的工作开发了针对细菌二肽基肽酶（DPP）和类胰蛋白酶的特异性检测方法，并且我们在龈沟液（GCF）中发现了它们。本研究的目的是确定在慢性牙周炎患者接受牙周治疗后它们的水平是否会降低。在25名未经治疗的患者的磨牙和前磨牙的近中颊侧位点测量探诊深度、探诊附着水平、牙龈指数、牙龈出血指数和菌斑指数。在第二次就诊时，用滤纸条收集龈沟液30秒。用牙周测量仪测定龈沟液体积，并将样本洗脱到缓冲液中。然后患者接受口腔卫生指导、龈上和龈下刮治以及其他适当的非手术治疗。四周后，从相同的16个位点收集龈沟液，并再次测量临床参数。在pH 8.0条件下，使用Ala-Pro-AFC通过荧光法测定类DPP活性，分别在加热至60℃ 30分钟和未加热的情况下进行。热敏感部分被视为细菌DPP活性。在pH 7.0条件下，使用Z-Val-Lys-Lys-Arg-AFC，加入2 mM二硫苏糖醇和10 μM Z-Phe-Ala-CH2测定细菌类胰蛋白酶活性。治疗后，临床参数、酶总活性和浓度均显著降低。使用个体患者或汇总患者数据，所有降低在患者和位点水平均具有统计学意义。细菌蛋白酶似乎反映了临床状况，可能在监测慢性炎症性牙周疾病中具有价值。
