Department of Oral and Maxillofacial Diseases, Institute of Dentistry, Helsinki University Central Hospital, University of Helsinki, Helsinki, Finland.
J Periodontal Res. 2010 Jun;45(3):353-60. doi: 10.1111/j.1600-0765.2009.01244.x. Epub 2009 Nov 9.
Emdogain (EMD), consisting mostly of amelogenin, is used in periodontal therapy to regenerate lost connective tissue. Emdogain is applied onto periodontally affected root surfaces, where it becomes exposed to proteolytic enzymes. In this study, we aimed to find out whether gingival crevicular fluid or matrix metalloproteinases (MMPs) could degrade EMD, and whether this degradation has consequences for in vitro cell proliferation.
We studied the effects of 156 gingival crevicular fluid samples collected from subjects with different stages of periodontal disease and from healthy control subjects and the effects of MMP-1, -2, -8, -9, -13 and -14 on the degradation of EMD using EMD-embedded zymography. The effects of gingival crevicular fluid with or without EMD and the effects of amelogenin on the proliferation of cultured periodontal ligament fibroblasts were studied by cell proliferation enzyme-linked immunosorbent assay kit.
Degradation of Emdogain induced by gingival crevicular fluid was greater in samples from all stages of periodontal diseases compared with healthy control samples. Of the MMPs studied, only MMP-2 and MMP-8 showed limited EMD-degrading activities. One hundred micrograms per millilitre of EMD increased proliferation of periodontal ligament fibroblasts on average by 24% (confidence interval 0.60-0.64) and at 200 microg/mL by 30% (confidence interval 0.62-0.68) compared with control fibroblasts (confidence interval 0.48-0.52). However, gingival crevicular fluid (10 microg/mL) together with 100 microg/mL EMD induced the proliferation only by 6% (confidence interval 0.51-0.55) and with 200 microg/mL EMD by 12% (confidence interval 0.54-0.58). Amelogenin at 200 microg/mL decreased the proliferation of periodontal ligament fibroblasts by 54% (confidence interval 0.22-0.25).
We suggest that diseased gingival crevicular fluid containing various proteases leads to degradation of EMD and decreased proliferation of periodontal ligament fibroblasts.
Emdogain（EMD）主要由釉原蛋白组成，用于牙周治疗以再生丢失的结缔组织。将 Emdogain 涂抹在牙周病受累的牙根表面，使其暴露于蛋白水解酶中。在这项研究中，我们旨在确定牙龈沟液或基质金属蛋白酶（MMPs）是否可以降解 EMD，以及这种降解是否会对体外细胞增殖产生影响。
我们研究了取自不同牙周病阶段的受试者和健康对照受试者的 156 份牙龈沟液样本的作用，以及 MMP-1、-2、-8、-9、-13 和 -14 对 EMD 降解的作用，使用 EMD 包埋的酶谱法。通过细胞增殖酶联免疫吸附试验试剂盒研究了含有或不含有 EMD 的牙龈沟液以及釉原蛋白对培养的牙周膜成纤维细胞增殖的影响。
与健康对照组相比，来自所有牙周病阶段的样本中 Emdogain 的降解程度更大。在所研究的 MMP 中，只有 MMP-2 和 MMP-8 显示出有限的 EMD 降解活性。100μg/mL 的 EMD 平均使牙周膜成纤维细胞增殖增加 24%（置信区间 0.60-0.64），200μg/mL 时增加 30%（置信区间 0.62-0.68），与对照成纤维细胞相比（置信区间 0.48-0.52）。然而，牙龈沟液（10μg/mL）与 100μg/mL 的 EMD 一起仅诱导增殖 6%（置信区间 0.51-0.55），与 200μg/mL 的 EMD 一起诱导增殖 12%（置信区间 0.54-0.58）。200μg/mL 的釉原蛋白使牙周膜成纤维细胞的增殖减少 54%（置信区间 0.22-0.25）。
我们认为含有各种蛋白酶的病变性牙龈沟液会导致 EMD 降解和牙周膜成纤维细胞增殖减少。
