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Immunoelectron Microscopic Study on Cytoskelcton of Pocket Epithelium and Infiltrating Cells

袋上皮及浸润细胞细胞骨架的免疫电镜研究

基本信息

批准号：
01571047
负责人：
HARA Yoshitaka
金额：
$ 1.34万
依托单位：
Nagasaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

It is easy to imagine that minute destruction of periodontal tissues precede clinical recognition of lesion. Considering that major causes of marginal periodontitis are plaque bacteria and the factors from them, the first region of destruction is presumably epithelial tissue. So I tried to clear the destruction of epithelial tissue, especially of its extracellular matrix. Further in this study extracellular matrix which consists of basement membrane, collagen type IV, laminin and fibronectin were researched. Materials of gingiva with periodontitis were obtained at periodontal surgeries and clinically healthy gingiva were used as controls. Frozen sections were made on routine way. Rabbit anti-collagen type IV, anti-laminin and anti-fibronectin were used as primary antibodies and goat anti-rabbit immunoglobulin conjugated with FITC as secondary antibodies. Then the sections were observed immunofluorecently. In addition infiltrating cells at unit square were counted immunohistologically. In clinically healthy gingiva collagen type IV and laminin were continuous on basement membrane and fibronectin was widely spreading among connective tissues. But when the amount of infiltrating cells increased, fibronectin disappeared and continuity of collagen type IV and laminin were lost. There was relationship between discontinuity and the kinds of infiltrating neutrophils and T-lymphocytes. Neutrophils have some proteinases, and in which matrix metalloproteinase to destroy the collagen type IV and laminin involved. So these results suggested that neutrophils have some relationship with minute destruction of periodontal tissue like discontinuity of basement membrane.

很容易想象，牙周组织的微小破坏先于临床识别病变。考虑到边缘性牙周炎的主要原因是牙菌斑细菌及其产生的因素，第一个破坏的区域可能是上皮组织。所以我试图清除上皮组织的破坏，特别是其细胞外基质的破坏。在这项研究中，进一步研究了由基底膜、IV 型胶原蛋白、层粘连蛋白和纤连蛋白组成的细胞外基质。牙周手术时获取患有牙周炎的牙龈材料，并以临床健康牙龈作为对照。按常规方法制作冰冻切片。使用兔抗IV型胶原、抗层粘连蛋白和抗纤连蛋白作为一抗，并使用与FITC缀合的山羊抗兔免疫球蛋白作为二抗。然后对切片进行免疫荧光观察。另外，对单位正方形的浸润细胞进行免疫组织学计数。在临床健康牙龈中，IV型胶原蛋白和层粘连蛋白在基底膜上连续，纤连蛋白广泛分布在结缔组织中。但当浸润细胞数量增加时，纤连蛋白消失，IV型胶原和层粘连蛋白的连续性丧失。间断性与浸润的中性粒细胞和T淋巴细胞的种类有关。中性粒细胞有一些蛋白酶，其中基质金属蛋白酶参与破坏IV型胶原蛋白和层粘连蛋白。因此，这些结果表明，中性粒细胞与牙周组织的微小破坏（例如基底膜的不连续性）有一定关系。