Bhide V M, Smith L, Overall C M, Birek P, McCulloch C A
Medical Research Council Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, ON.
J Periodontol. 2000 May;71(5):690-700. doi: 10.1902/jop.2000.71.5.690.
Quantification of gingival crevicular fluid matrix metalloproteinase activity may provide improved assessment of periodontal disease status and response to treatment. A fluorogenic matrix metalloproteinase substrate assay (FSA) has been developed using a methoxycoumarin-containing septapeptide analog of the alpha2(I) collagen cleavage site. This substrate exhibits increased fluorescence following cleavage by many matrix metalloproteinases, and the enzyme activity can be readily estimated with a fluorimeter. Here we compared this assay with classical methods of periodontal assessment including bleeding on probing, crevicular fluid flow, and probing depth to assess its utility as an indicator of changes in periodontal status and treatment response.
Complete measurements of probing depth were obtained for Ramfjord teeth on subjects who had been previously treated for periodontitis. Subjects were subsequently divided into groups based on existing periodontal disease severity: gingivitis (n = 21), stable periodontitis (n = 41), and severe periodontitis (n = 50). Crevicular fluid volume, bleeding on probing, and FSA were measured at each Ramfjord tooth or substitute. After baseline measurements, subjects received subgingival scaling and prophylaxis; 3 months later, they were reassessed.
FSA measurements were positively associated with severity of disease at baseline. After treatment there were substantial reductions of FSA in gingivitis (approximately 51%; P <0.01) and severe periodontitis (approximately 45%; P <0.001), but not in stable periodontitis (13%; P >0.2). All groups showed a positive association between FSA measurements and higher bleeding scores at individual sites. FSA measurements were also positively associated with crevicular fluid flow at baseline, but after treatment there was a approximately 67% decrease (P <0.01) in the highest crevicular fluid flow class. There were significant reductions of FSA at follow-up for sites with probing depths between 0 to 3 mm (23%; P <0.05) and 4 to 6 mm (31%; P <0.05). However, the largest reduction was for sites with probing depth between 7 to 9 mm (49%; P <0.001).
These results indicate that monitoring patients by measurement of matrix metalloproteinase levels in gingival crevicular fluid with the quenched fluorescent substrate assay provides estimates of inflammatory status, periodontal destruction, and response to treatment, especially in more severe periodontitis lesions.
龈沟液基质金属蛋白酶活性的定量分析可能有助于更好地评估牙周疾病状态及对治疗的反应。一种基于含甲氧基香豆素的α2(I)胶原裂解位点七肽类似物的荧光基质金属蛋白酶底物检测法（FSA）已被开发出来。该底物在被多种基质金属蛋白酶裂解后荧光增强，酶活性可用荧光计轻松测定。在此，我们将该检测法与牙周评估的经典方法（包括探诊出血、龈沟液流量和探诊深度）进行比较，以评估其作为牙周状态变化和治疗反应指标的效用。
对曾接受过牙周炎治疗的受试者的Ramfjord牙进行完整的探诊深度测量。随后根据现有的牙周疾病严重程度将受试者分为几组：牙龈炎（n = 21）、稳定期牙周炎（n = 41）和重度牙周炎（n = 50）。在每颗Ramfjord牙或替代牙上测量龈沟液量、探诊出血情况及FSA。在进行基线测量后，受试者接受龈下刮治和预防性治疗；3个月后，再次对他们进行评估。
FSA测量值在基线时与疾病严重程度呈正相关。治疗后，牙龈炎组（约51%；P <0.01）和重度牙周炎组（约45%；P <0.001）的FSA大幅降低，但稳定期牙周炎组未降低（13%；P >0.2）。所有组在各个部位的FSA测量值与更高的出血评分之间均呈正相关。FSA测量值在基线时也与龈沟液流量呈正相关，但治疗后最高龈沟液流量组下降了约67%（P <0.01）。对于探诊深度在0至3毫米（23%；P <0.05）和4至6毫米（31%；P <0.05）的部位，随访时FSA有显著降低。然而，探诊深度在7至9毫米的部位下降幅度最大（49%；P <0.001）。
这些结果表明，通过使用淬灭荧光底物检测法测量龈沟液中的基质金属蛋白酶水平来监测患者，能够评估炎症状态、牙周破坏情况及对治疗的反应，尤其是在更严重的牙周炎病变中。
