MULTIPLE ANALYSIS OF PERIODONTAL LESIONS BY DIFFERENTIAL DISPLAY

2002 Fiscal Year Final Research Report Summary

• Project/Area Number: 13672185
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Periodontal dentistry
• Research Institution: OSAKA UNIVERSITY
• Principal Investigator: KITAMURA Masahiro OSAKA UNIVERSITY, DENTAL HOSPITAL, ASSISTANT PROFESSOR, 歯学部附属病院, 講師 (10243247)
• Co-Investigator(Kenkyū-buntansha): SAHO Teruyuki OSAKA UNIVERSITY, DENTAL HOSPITAL, Instructor, 歯学部附属病院, 助手 (10263295) ; IKEZAWA Kazuhiko OSAKA UNIVERSITY, DENTAL HOSPITAL, ASSISTANT PROFESSOR, 歯学部附属病院, 講師 (80294114) ; MURAKAMI Shinya OSAKA UNIVERSITY, GRADUATE SCHOOL OF DENTISTRY, PROFESSOR, 大学院・歯学研究科, 教授 (70239490)
• Project Period (FY): 2001 – 2002
• Keywords: penodonfad disease / gingival tissue / inflammatory cytokine / growth factor / cDNAmicroarray / gene expression

Research Abstract

This study was designed to investigate the largescale gene expression of inflammatory cytokines, growth factors and enzymes involved periodontal tissue destruction by CDNA microarray for diagnosis of periodontal diseases. We selected about 100 known genes for arrayed targets, contained inflammatory cytokines, growth factors and enzymes these were closely related with pathogenesis of periodontal diseases. Based on the information of the gene expression profile of human periodontal ligament (PDL), we added about 1,100 nonredundant clones for arrayed targets from 3'cDNA library ofPDL. PCR was performed to amplify cDNAs for the targets and about 1,200 species of cDNA were printed onto a polyLlysine coated slide glass using the microarrayer. In order to verify the CDNA microarray for diagnosis of eriodontal diseases, we isolated RNA from PDL.cells cultured at different time points and RNAs were reversetranscribed and differentially labeled with Cy3 and Cy5, and hybridized onto the microarray. In result, we detected the differential expression ofseveral genes during the cytodifferentiation of PDL cells. Furthermore, we applied this cDNA microarray to the largescale gene expression analysis in periodontal tissue. Total RNA was extracted from the gingival tissue sampled at periodontal surgery and human gingival fibrobrast (HGF) cultured in vitro. The RNAs were reverse transcribed and differentially labeled with Cy3 and Cy5, and hybridized onto the microarray. In the gingival tissue, we detected upregulation of the gene expressions involved immunocompetent cells, such as immunogloburin genes and chemokine genes. In another way, the upregulation of the genes of collagen and cell adhesion molecules was detected in HGF. Present results show that the cDNA microarray developed in this study is a powerful tool to analyze the largescale gene expression in periodontal tissue and suggest that there is a possibility of multiple analysis for periodontal disease condition by this CDNA