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

在本研究中，我们试图建立诊断牙周病分子发病机制的策略，基因组聚合酶链反应（PCR）技术使我们能够灵敏、特异地检测龈下牙菌斑中的牙龈卟啉单胞菌（P.g.）和伴放线放线菌（A.a.）。此外，利用逆转录（RT）-PCR技术，建立了分离牙龈标本中多种细胞因子mRNA的半定量检测体系。然后，我们收集了成人和早发性牙周炎（EOP）患者在首次就诊、重新评估和支持性牙周治疗（SPT）时的龈下牙菌斑、牙龈标本和血清，并通过分子生物学技术对这些样本进行了分析。如上所述，我们观察到发炎的牙周病变中多种炎症细胞因子的 mRNA 表达上调。特别是，大约 50% 的 EOP 病变部位被分为 IL-8 和 IFN-D2γD2 mRNA 表达相对较低的组。在另一项研究中，检测了抗 P.g. 的 IgG 亚类抗体的血清水平。接受SPT 5年的牙周炎患者牙槽骨丢失情况显示，IgG2水平与△骨丢失评分之间存在统计学上显着的正相关关系。 (p<0.001) 这表明牙周治疗后观察到的延长的 IgG2 反应可能与复发性或持续性牙周破坏有关。在体外研究中，我们证明牙龈上皮细胞优先分泌 IL-8 和 MCP-。 1 当用 P.g. 声波提取物刺激时，我们还阐明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中炎症细胞因子的表达。这些结果表明，这些常驻细胞在发炎的牙周病变中积极参与细胞因子网络。

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, 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。

H.Okada, S.Murakami: "Cytokine expression in periodontal health and disease" Crit.Rev.Oral Biol.Med.(in press). (1998)

H.Okada、S.Murakami：“牙周健康和疾病中的细胞因子表达”Crit.Rev.Oral Biol.Med.（出版中）。

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。