Establishment of molecular biological diagnostic strategies for periodontal diseases

牙周病分子生物学诊断策略的建立

• 批准号: 09307043
• 负责人: OKADA Hiroshi
• 金额: $ 16.45万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09307043/
• 关键词: PERIODONTITIS; DIAGNOSIS; CYTOKINES; PCR; PERIODONTOPATHIC BACTERIA; SERUM ANTIBODY; GINGIVAL EPITHELIAL CELL; GINGIVAL FIBROBLAST; mRNA; RT-PCR; 分子生物学的診断; T細胞; PCR; ELISA; 炎症

In this study, we attempted to establish the strategies to diagnose the molecular pathogenesis of periodontal diseases. Genomic polymerase chain reaction (PCR) technique enabled us to sensitively and specifically detect Porphyromonas gingivalis (P.g.) and Actinobucillus actinomycetemcomitans (A.a.) in subgingival dental plaques. Furthermore, utilizing reverse transcription (RT)-PCR technique, we established the semiquantitative detection system of multiple cytokine mRNA in gingival specimens isolated by needle biopsy method. We then collected subgingival dental plaque, gingival specimens and serum from adult and early-onset periodontitis (EOP) patients at the times of first visit, reevaluation and supportive periodontal therapy (SPT) and analyzed those samples by molecular biological techniques described above. We observed the upregulated mRNA expression of a variety of inflammatory cytokines in inflamed periodontal lesions. Interestingly, we could statistically divide the multiple cyt … More okine mRNA expression profiles into 5 groups by cluster analysis. In particular, approximately 50% of the diseased sites from EOP were categorized into the groups showing relatively low expression of IL-8 and IFNィイD2γィエD2 mRNA.In another study, serum levels of IgG subclass antibodies against P.g. were examined in relation to alveolar bone loss in periodontitis patients receiving SPT for 5 years. Statistically significant positive correlation was seen for the relationship between IgG2 levels and △bone loss score (p<0.001) in the SPT patients. This suggests that the prolonged IgG2 response observed after periodontal treatment may be associated with recurrent or persistent periodontal destruction.In in vitro studies, we demonstrated that gingival epithelial cells preferentially secreted IL-8 and MCP-1 when stimulated with P.g. sonic extract. In addition, we clarified that adhesive interactions between lymphocytes and gingival fibroblasts induced inflammatory cytokine expression in the gingival fibroblasts. These results suggest that those resident cells actively participate in the cytokine network in inflamed periodontal lesions. Less

本研究试图建立牙周病分子发病机制的诊断策略。基因组聚合酶链反应（PCR）技术使我们能够敏感和特异地检测牙龈卟啉单胞菌（P.g.）和伴放线杆菌Actinobucillus actinomycetemcomitans（A.a.）龈下牙菌斑中。利用逆转录-聚合酶链反应（RT-PCR）技术，建立了牙龈组织中多种细胞因子mRNA的半定量检测体系。然后，我们在首次就诊、重新评估和支持性牙周治疗（SPT）时收集了成年和早发性牙周炎（EOP）患者的龈下牙菌斑、牙龈标本和血清，并通过上述分子生物学技术分析了这些样本。我们观察到炎症牙周病变中多种炎性细胞因子的mRNA表达上调。有趣的是，我们可以从统计学上将多个细胞周期 ...更多信息 通过聚类分析将okine mRNA表达谱分为5组。特别地，来自EOP的大约50%的患病部位被分类为显示相对低表达的IL-8和IFN γ D2γ D2 mRNA的组。在接受SPT 5年的牙周炎患者中，检查与牙槽骨丢失的关系。SPT患者IgG 2水平与△骨丢失评分呈显著正相关（p<0.001）。这表明，牙周治疗后观察到的延长的IgG 2反应可能与复发性或持续性的牙周damage.In体外研究中，我们证明，牙龈上皮细胞优先分泌IL-8和MCP-1时，刺激P.g.声波提取此外，我们阐明了淋巴细胞和牙龈成纤维细胞之间的粘附相互作用诱导牙龈成纤维细胞中的炎性细胞因子表达。这些结果表明，这些常驻细胞积极参与炎症牙周病变的细胞因子网络。少

项目成果

村上伸也: "医歯薬出版"歯周病診断のストラテジー. 246 (1999)

村上伸也：《石药出版社》牙周病诊断策略246（1999）。

村上伸也: "ヒアルロン酸の生物学的活性を考える"日本炎症学会雑誌. 19. 307-318 (1999)

Shinya Murakami：“考虑透明质酸的生物活性”日本炎症学会杂志 19. 307-318 (1999)。

S. Murakami and H. Okada: "Diagnosis of periodontal disease based on the techniques of molecular biology."The Journal of the Japan Demtal Association. 16. 6-13 (1997)

S. Murakami 和 H. Okada：“基于分子生物学技术的牙周病诊断”。日本牙科协会杂志。

S. Murakami and T. Nozaki: "Diagnostic strategies of periodontal diseases"Ishiyaku-shuppan. 158-167 (1999)

S. Murakami 和 T. Nozaki：“牙周病的诊断策略”Ishiyaku-shuppan。

S. Murakami, Y. Shimabukuro and H. Okada: "Biological activities of hyaluronan"Japanese Joumal of Inflammation. 19. 307-318 (1999)

S. Murakami、Y. Shimabukuro 和 H. Okada：“透明质酸的生物活性”日本炎症杂志。