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Investigation of mechanism of periodontal destruction in rat experimental periodontitis caused by topical application of lipopolysaccharide

局部应用脂多糖致大鼠实验性牙周炎牙周破坏机制的探讨

基本信息

批准号：
06454510
负责人：
IJUHIN Naokuni
金额：
$ 4.61万
依托单位：
OSAKA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

To elucidate histopathological mechanism of periodontal ligament destruction in the marginal periodontitis caused by lipopolysaccharide, (1) we have firstly tried to investigate the collagen-phagocytotic and/or collagenolytic activities of periodontal ligament fibroblasts using tissue culture cells with histochical and immunohistochemical methods. We could establish the fibroblastic cell lines from rat molar periodontat ligaments. These cells were polyhedral, round or spindle in shape on the bottom surface of tissue culture plate. Histochemically and immunohistochemically, they showed characteristics of periodontal ligament fibroblasts, such as positive reaction of alkaline phosphatase and alizarin red staining for calcification, immunostaining for vimentin and type I collagen. They also revealed slight immunohistochemical positivities for keratin, osteocalcine and OX-43 (marker for endothelial cell and macrophage). We cultured these fibroblasts three-dimensionally in the collagen gel … More and tried to demonstrate phagocytosis of collagen by the culture cells under the electron microscopic observation. But, it was difficult to find the ingested collagen fibrils in the cultured cells even under the ultrastructural level. Trial to demonstrate the phagicytosed collagen immunohisto-chemically in the cultured fibroblastic cells was also unsuccessful, as it was difficult to differentiate the ingested collagen fibrils from the synthesized collagen in the cells. On the other hand, (2) we studied dynamics of macrophages in the rat experimental periodontitis caused by topical application of lipopolysaccharide, because synthesis of collagenase and/or IL-1beta by macrophage is suspected to cause destruction of periodontal ligament. As a result, it was revealed that ED1 positive exudative tissue macrophages from blood monocytes increased in the inflammatory periodontal lesion after LPS-administration. These macrophages produced IL-1beta and was suggested to participate destruction of periodontal tissue. Ia antigen positive dendritic cells also increased in the lesion after LPS-administration, which suggested that much of these macrophages changed to antigen presenting cells and might have important role for reactivity of local immune response. Less

为了阐明脂多糖引起的边缘性牙周炎牙周膜破坏的组织病理学机制，（1）我们首先尝试使用组织培养细胞通过组织学和免疫组织化学方法研究牙周膜成纤维细胞的胶原吞噬和/或胶原溶解活性。我们可以从大鼠磨牙牙周韧带中建立成纤维细胞系。这些细胞在组织培养板的底面上呈多面体、圆形或纺锤形。组织化学和免疫组织化学表现出牙周膜成纤维细胞的特征，如碱性磷酸酶和钙化茜素红染色阳性，波形蛋白和I型胶原免疫染色。他们还揭示了角蛋白、骨钙素和 OX-43（内皮细胞和巨噬细胞的标记物）的轻微免疫组织化学阳性。我们在胶原凝胶中对这些成纤维细胞进行三维培养，并试图在电子显微镜观察下证明培养细胞对胶原的吞噬作用。但是，即使在超微结构水平下，也很难在培养细胞中找到摄入的胶原纤维。用免疫组织化学方法证明培养的成纤维细胞中吞噬的胶原蛋白的试验也不成功，因为很难区分摄入的胶原纤维和细胞中合成的胶原蛋白。另一方面，(2)我们研究了局部应用脂多糖引起的大鼠实验性牙周炎中巨噬细胞的动态，因为巨噬细胞合成胶原酶和/或IL-1β被怀疑会导致牙周膜的破坏。结果表明，给予LPS后，炎症性牙周病灶中来自血液单核细胞的ED1阳性渗出组织巨噬细胞增加。这些巨噬细胞产生 IL-1β，并被认为参与牙周组织的破坏。给予LPS后，病灶中Ia抗原阳性树突状细胞也有所增加，这表明这些巨噬细胞中的大部分转变为抗原呈递细胞，并且可能对局部免疫反应的反应性具有重要作用。较少的