Detection of bacterial species using 16S rDNA existed in deep layers of carious lesions with pulp exposure embedded with plastic material and analysis of the pathogenesis of pulpitis

16S rDNA检测牙髓暴露包埋塑料材料龋损深层细菌种类及牙髓炎发病机制分析

• 批准号: 13672000
• 负责人: OZAKI Kazumi
• 金额: $ 2.24万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672000/
• 关键词: Dentinal caries; 16S rDNA; Embedding material for TEM; TEM-in situ hybridization; RT-PCR; Toll-like receptor

Resin-embedded human carious dentin was used as template for polymerase chain reaction (PCR) detection of bacterial 16 rDNA existed in deep layers of carious lesions. A total of 34 bacterial spesies were detected from 39 human carious samples. 4.8 bacterial species per carious sample of coronal caries, 3.9 species of root caries were detected.Characteristics of Toll-like receptor (TLR)-expressing cells in human dental pulp were studied. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis comfirmed the expression of TLR-2 Mrna in all of the inflamed pulps. In situ hybridization analysis using digoxigenin-labeled single strand Crna probe showed that TLR-2, 4 and 9 Mrna-expressing cells could be found in the inflamed pulps. Immunohistological analysis at a light microscopic level showed that TCR-2 and 4 protein in the cell body. In antigen-presenting cell (APC), dendritic cells were expressed TLR-2 protein, and macrophage-like cells were expressed TLR-4 protein in the cell body. These results suggested that TLR might be involved in the pathogenesis of pulpitis.

以树脂包埋的人龋齿牙本质为模板，采用聚合酶链反应（PCR）检测龋损深层细菌16 rDNA。从 39 份人类龋齿样本中总共检测到 34 种细菌。每个冠状龋齿样品检出4.8种细菌，检测出3.9种根龋。研究了人牙髓中Toll样受体（TLR）表达细胞的特征。逆转录聚合酶链反应 (RT-PCR) 分析证实了所有发炎牙髓中 TLR-2 Mrna 的表达。使用地高辛标记的单链 Crna 探针进行原位杂交分析表明，在发炎的牙髓中可以发现表达 TLR-2、4 和 9 Mrna 的细胞。光镜水平的免疫组织学分析显示细胞体内存在TCR-2和4蛋白。在抗原呈递细胞(APC)中，胞体内树突状细胞表达TLR-2蛋白，巨噬细胞样细胞表达TLR-4蛋白。这些结果表明TLR可能参与牙髓炎的发病机制。

项目成果

Ozaki K, et al.: "Histological observation of bacterial invasion at an electron microsdopic level"Jpn J Conserv Dent. 45 (special issue). 46-46 (2002)

Ozaki K 等人：“电子显微镜水平上细菌入侵的组织学观察”Jpn J Conserv Dent。

Matsuo T, et al.: "An immunohistological study of the localization of bacteria invading root pulpal walls of teeth with periapical lesions"J. Endodontics. in press. (2002)

Matsuo T 等人：“侵入根尖周病变牙齿根髓壁的细菌定位的免疫组织学研究”J.

Suenobu S, et al.: "Propwrties of Glass-Ionomer Luting Cements containing really spherical silicaparticle coated with silver"Jpn Conserv Dent. 44 (special issue). 103-103 (2001)

Suenobu S 等人：“含有银涂层的真球形二氧化硅颗粒的玻璃离聚物润滑水泥的特性”Jpn Conserv Dent。