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.

采用聚合酶链反应（PCR）技术，以树脂包埋的人龋坏牙本质为模板，检测龋坏深层的细菌16 rDNA。从39份人龋标本中检出34种细菌。4.8对人牙髓组织中Toll样受体（TLR）表达细胞的特征进行了研究。逆转录聚合酶链反应（RT-PCR）分析证实TLR-2 mRNA的表达在所有的炎症牙髓。地高辛标记的单链Crna探针原位杂交分析表明，TLR-2，4和9 mRNA表达细胞可以在发炎牙髓中找到。免疫组化光镜下可见TCR-2和TCR-4蛋白在细胞内表达。在抗原提呈细胞（APC）中，树突状细胞表达TLR-2蛋白，巨噬细胞样细胞表达TLR-4蛋白。提示TLR可能参与牙髓炎的发病过程。

项目成果

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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)

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