Mailhot J M, Potempa J, Stein S H, Travis J, Sterrett J D, Hanes P J, Russell C M
Department of Periodontics, Medical College of Georgia, Augusta 30912, USA.
J Clin Periodontol. 1998 Jul;25(7):578-84. doi: 10.1111/j.1600-051x.1998.tb02491.x.
The objective of this research was to determine the effectiveness of a biochemical assay which measures proteolytic enzyme activity in gingival crevicular fluid (GCF) and to relate this enzyme activity to clinical parameters traditionally utilized for periodontitis detection. A clinical trial was conducted on 8 periodontitis subjects with > or =4 sites exhibiting a loss of attachment of > or =5 mm and probing depths of > or =5 mm with bleeding on probing. On each subject, a plaque index was performed, followed by GCF sampling at those sites which exhibited a loss of attachment and probing depths. GCF was analyzed for activity against benzoyl-L-arginine-p-nitroanilide in the presence (BAPNA w/gly-gly) and the absence (BAPNA w/o gly-gly) of glycyl-glycine and against MeOSuc-Ala-Ala-Pro-Val-pNA and Suc-Ala-Ala-Pro-Phe-pNA for neutrophil serine proteinases activity (elastase and cathepsin G, respectively). Subsequently, a gingival index was performed, attachment levels and probing depths were recorded using a constant force probe with bleeding on probing being noted. A split-mouth design was employed and half mouths were randomly assigned to the following treatment groups: group A, half of the mouth received scaling/root planing and polishing: group B, half of the mouth received no treatment (control). Subjects were treated, then instructed on toothbrushing and interdental cleaning. After 4 weeks, subjects returned to receive a plaque index; GCF sampling, gingival index, attachment levels, probing depths and bleeding on probing as described above. Using a paired Student t-test, the findings suggest that BAPNA w/gly-gly was significantly less in treatment sites than in non-treated control sites (p=0.05). No such correlation was found for other activities, including neutrophil serine proteinases which were shown to occur in GCF in free, proteolytically active forms. In addition, significant treatment effects were detected for probing depths (p= 0.03) which reduced by 1.3 mm and attachment levels (p=0.02) which gained 0.7 mm. The reduction of P. gingivalis from treated periodontitis sites as detected by a significant decrease in BAPNA w/ gly-gly may prove to be a valuable marker for periodontal disease activity.
本研究的目的是确定一种生化检测方法的有效性，该方法用于测量龈沟液（GCF）中的蛋白水解酶活性，并将这种酶活性与传统上用于检测牙周炎的临床参数相关联。对8名牙周炎患者进行了一项临床试验，这些患者有≥4个位点出现≥5 mm的附着丧失，探诊深度≥5 mm且探诊出血。对每位受试者进行菌斑指数检测，然后在那些出现附着丧失和探诊深度的位点采集龈沟液样本。分析龈沟液在存在甘氨酰甘氨酸（BAPNA w/gly-gly）和不存在甘氨酰甘氨酸（BAPNA w/o gly-gly）的情况下对苯甲酰-L-精氨酸-对硝基苯胺的活性，以及对甲磺酰基琥珀酰-丙氨酰-丙氨酰-脯氨酰-缬氨酰-对硝基苯胺和琥珀酰-丙氨酰-丙氨酰-脯氨酰-苯丙氨酰-对硝基苯胺的中性粒细胞丝氨酸蛋白酶活性（分别为弹性蛋白酶和组织蛋白酶G）。随后，进行牙龈指数检测，使用恒力探针记录附着水平和探诊深度，并记录探诊出血情况。采用半口设计，将半口随机分配到以下治疗组：A组，半口接受龈上洁治/根面平整和抛光；B组，半口不接受治疗（对照组）。对受试者进行治疗，然后指导他们刷牙和使用牙间隙清洁工具。4周后，受试者返回接受菌斑指数检测；如上述进行龈沟液采样、牙龈指数检测、附着水平、探诊深度和探诊出血情况检测。使用配对学生t检验，结果表明治疗位点的BAPNA w/gly-gly显著低于未治疗的对照位点（p = 0.05）。对于其他活性，包括以游离、蛋白水解活性形式存在于龈沟液中的中性粒细胞丝氨酸蛋白酶，未发现这种相关性。此外，检测到探诊深度有显著治疗效果（p = 0.03），探诊深度减少了1.3 mm，附着水平有显著治疗效果（p = 0.02），附着水平增加了0.7 mm。通过BAPNA w/ gly-gly的显著降低检测到治疗的牙周炎位点中牙龈卟啉单胞菌的减少，这可能被证明是牙周疾病活动的一个有价值的标志物。
