Immunoelectron Microscopic Study on Cytoskelcton of Pocket Epithelium and Infiltrating Cells

很容易想象，牙周组织的微小破坏先于病变的临床识别。考虑到边缘性牙周炎的主要原因是菌斑细菌和它们的因素，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，特别是它的细胞外基质。本研究进一步研究了由基底膜、IV型胶原、层粘连蛋白和纤连蛋白组成的细胞外基质。牙周炎牙龈材料在牙周手术中获得，临床健康牙龈作为对照。常规冰冻切片。兔抗IV型胶原、抗层粘连蛋白和抗纤连蛋白作为一抗，羊抗兔免疫球蛋白与FITC偶联作为二抗。然后对切片进行免疫荧光观察。此外，还对单位正方形的浸润细胞进行了免疫组织学计数。在临床健康的牙龈IV型胶原和层粘连蛋白连续的基底膜和纤维连接蛋白广泛传播的结缔组织。但随着浸润细胞数量的增加，纤维连接蛋白消失，IV型胶原和层粘连蛋白失去连续性。非连续性与浸润的中性粒细胞和T淋巴细胞的种类有关。中性粒细胞中有一些蛋白酶，其中基质金属蛋白酶参与破坏IV型胶原和层粘连蛋白。提示中性粒细胞与牙周组织的微小破坏如基底膜断裂有一定关系。

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.