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Molecular-biological analysis by DNA microarray for diagnosis of occlusal trauma

DNA 微阵列分子生物学分析用于诊断咬合创伤

基本信息

批准号：
15390646
负责人：
KITAMURA Masahiro
金额：
$ 9.34万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390646/
关键词：
DNA microarray Occlusal trauma Periodontal diseases

项目摘要

This study was designed to investigate the mechanisms involved occurrence of occlusal trauma and search the diagnostic makers of occlusal trauma by cDNA microarray and semi-measurable reverse transcription-polymerase chain reaction (RT-PCR). In the in vitro occlusal trauma model utilizing cell-stretcher (Scholertec NS-200), we detected up-regulation of the gene expressions in the stretched human periodontal ligament cell (HPDL), such as cyclooxygenase-2 (Cox-2), c-fos, integrin β-1, β-actin and adenosine deaminase genes. In another way, the genes associated with periodontal ligament, such as periodontal ligament associated protein-1 (PLAP-1) and periostin, did not showed the change of gene expressions by the mechanical stress. Furthermore, in order to clarify the diagnostic makers of occlusal trauma, we detected the mRNA of inflammatory cytokines and mediators by RT-PCR in the gingival tissues with occlusal trauma and without occlusal trauma collected by needle-punched biopsy methods. In result, IL-1β, IL-6, IL-8 and Cox2 mRNA expression were decreased and IFN-γ mRNA expression was elevated in the periodontal lesions decreased the occlusal trauma by occlusal adjustment. In another way, the mRNA of inflammatory cytokines and mediators detected in this study did showed no change or slight elevation of the expression in the periodontal lesions with occlusal trauma treated with no occlusal adjustment. Present results show that the mechanical stress promotes the various gene expressions associated with inflammatory reactions and bone destruction in HPDL and suggest that the inflammatory cytokines and mediators detected in this study, such as IL-1β, IL-6, IL-8 and Cox2 and IFN-γ, could be the potential diagnostic makers of occlusal trauma.

本研究旨在通过基因芯片和半定量逆转录聚合酶链式反应(RT-PCR)研究咬合创伤的发生机制，寻找咬合创伤的诊断标记物。在体外咬合创伤模型中，我们检测了人牙周韧带细胞的环氧合酶-2(COX-2)、c-fos、整合素β-1、β-肌动蛋白和腺苷脱氨酶基因的表达上调。与牙周膜相关的基因如牙周膜相关蛋白-1(PLAP-1)和骨膜蛋白(Periostin)等在机械应力作用下没有发生基因表达的变化。此外，为了明确咬合创伤的诊断标记物，我们用RT-PCR方法检测了有咬合创伤和无咬合创伤的牙周组织中炎症细胞因子和介质的mRNA的表达。结果表明，牙周病变组织中IL-1、IL-6、IL-8和COX2mRNA的表达降低，而干扰素-γ的表达升高，减轻了咬合调整对咬合创伤的影响。另一方面，本研究中检测到的炎性细胞因子和介质的mRNA在未进行咬合调整的牙周损伤中的表达没有变化或略有升高。目前的结果表明，机械应激促进了与炎症反应和骨破坏相关的各种基因的表达，提示本研究检测到的炎性细胞因子和介质，如IL-1、β、IL-6、IL-8、COX_2和干扰素-γ，可能是咬合创伤的潜在诊断指标。