Investigation of effect of lipopolysaccharide on phagocytic activity of collagen fibrils by periodontal fibroblast

脂多糖对牙周成纤维细胞吞噬胶原纤维活性的影响研究

• 批准号: 03670846
• 负责人: IJUHIN Naokuni
• 金额: $ 1.28万
• 依托单位: OSAKA UNIVERSITY (1993)Hiroshima University (1991-1992)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670846/
• 关键词: Lipopolysaccharide; Periodontium; Fibroblast; Collagen fibril; Phagocytosis; Rat; Tissue culture; Morphometry; 貪食; コラ-ゲン原線維

To investigate the effect of lipopolysaccharide on phagocytic activity of collagen fibrils by periodontal fibroblasts, We studied rat molar gingival connective tissue and periodontal ligament under light and electron microscopy after topical application of LPS (5 mg/ml in physiological salt solution, LPS from Escherichia coli from Sigma Chemical Co., St.Louis, MO USA) on the gingival sulcus. Collagen phagocytosis by gingival and periodontal fibroblasts was studied by enzyme histochemistry and morphometric analysis. Phagocytic activity of collagen fibrils by fibroblasts was evaluated by counting the number of collagen-containing vacuoles inside fibroblasts that were present within a defined area (1200mum^2). Values obtained from fibroblasts in the subepithelial connective tissue, the region near the alveolar crest, and the middle region of periodontal tissue were compared. Periodontal ligament fibroblasts showed increased phagocytosis of the collagen fibrils from 3 hours to 1 day after … More topical application of LPS,but, no differences were observed in the gingival tissue. The intracytoplasmic vacuoles containing collagen fibrils were of various sizes and shapes, and showed positive for acid phosphatase and/of alkaline phosphatase reaction. These results indicated that endotoxin may enhance the the degradation of collagen by stimulating the phagocytotic activity of the periodontal ligament fibroblasts. To study the effect of endotoxin on phagocytic activity of collagen fibrils by periodontal fibroblast in vitro system, we obtained 4 fibroblastic spindle-shaped cell lines derived from rat molar periodontal ligaments. These cells were spindle to cuboidal in shape, and showed positivities for alkaline phosphatase reaction and alizarin red S stain for calcium, which are characteristics of periodontal ligament fibroblasts. These cells were cultivated in the collagen gel, but, collagen phagocytosis by these cells were not seen under the ultrastructural observation. We are further studying to get good results, changing a sort of medium, concentrations of collagen gel, methods of observation, etc. Less

为了研究脂多糖对牙周成纤维细胞吞噬胶原原纤维活性的影响，我们在光镜和电镜下研究了LPS（生理盐溶液5 mg/ml，来自Sigma Chemical Co., St.Louis, MO . USA）外用的LPS（大肠杆菌LPS）对大鼠牙周沟的结缔组织和牙周韧带。采用酶组织化学和形态计量学方法研究牙龈和牙周成纤维细胞对胶原的吞噬作用。通过计算在一定范围内（1200mum^2）成纤维细胞内含有胶原蛋白的空泡的数量来评估成纤维细胞对胶原原纤维的吞噬活性。比较了上皮下结缔组织、牙槽嵴附近区域和牙周组织中部区域的成纤维细胞所获得的数值。局部应用LPS后3小时至1天，牙周韧带成纤维细胞对胶原原纤维的吞噬作用增加，但对牙龈组织的吞噬作用无明显影响。含有胶原原纤维的胞浆内空泡大小形状不一，酸性磷酸酶和碱性磷酸酶反应阳性。这些结果表明，内毒素可能通过刺激牙周韧带成纤维细胞的吞噬活性来促进胶原蛋白的降解。为了研究内毒素对体外培养的牙周成纤维细胞吞噬胶原原纤维活性的影响，我们获得了4株大鼠磨牙牙周韧带成纤维梭形细胞株。细胞呈梭形至立方状，碱性磷酸酶反应阳性，茜素红S染色阳性，具有牙周韧带成纤维细胞的特征。细胞在胶原凝胶中培养，超微结构观察未见细胞吞噬胶原。为了取得良好的效果，我们正在进一步研究，改变一种培养基、胶原凝胶的浓度、观察方法等。少

项目成果

高田 隆: "ラット臼歯部歯周靱帯由来初代培養細胞の形態学的観察." 日本歯周病学会会誌. 34. 19- (1992)

Takashi Takada：“大鼠磨牙牙周膜原代培养细胞的形态观察”，日本牙周病学会杂志 34. 19- (1992)。

Naokuni Ijuhin: "Enhanced collagen phagocytosis by rat molar periodontal fibro blasts after topical application of lipopolysaccharide-Ultra structural observations and morphometric analysis." Journal of periodontal Research. 27. 167-175 (1992)

Naokuni Ijuhin：“局部应用脂多糖后大鼠磨牙牙周成纤维细胞对胶原蛋白的吞噬作用增强 - 超结构观察和形态分析。”

Takashi Takata: "Morphological observation of explant tissue culture cells derived from rat molar periodontal ligament fibroblasts" Journal of Japanese Society of Periodontology. 34. 19 (1992)

Takashi Takata：“来自大鼠磨牙牙周膜成纤维细胞的外植体组织培养细胞的形态观察”日本牙周病学会杂志。

高田 隆: "ラット臼歯部歯周靱帯由来初代培養細胞の形態学的観察.(2)コラーゲン・ゲル包埋培養法による観察" 日本歯周病学会会誌. 35. 153- (1993)

Takashi Takada：“来自大鼠磨牙牙周膜的原代培养细胞的形态观察。(2)胶原凝胶包埋培养法的观察”日本牙周病学会杂志35. 153-(1993)。

Naokuni Ijuhin: "Enhanced collagen phagocytosis by rat molar periodontal fibroblasts after topical application of lipopolysaccharide-Ultrastructural observations and morphometric analysis" Journal of Periodontal research. 21. 167-175 (1992)

Naokuni Ijuhin：“局部应用脂多糖后大鼠磨牙牙周成纤维细胞对胶原蛋白的吞噬作用增强 - 超微结构观察和形态分析”牙周研究杂志。