Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
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A study on the osteoblast differentiation using osteocalcin gene promoter controlling luciferase expression

리포터유전자를 이용한 조골세포 분화정도에 관한 연구

  • Kim, Kyoung-Hwa (Dept. of Periodontology, BK21 Craniomaxillofacial life science) ;
  • Park, Yoon-Jeong (Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University) ;
  • Lee, Yong-Moo (Dept. of Periodontology, School of Dentistry, Seoul National University) ;
  • Han, Jung-Suk (Dept. of Prosthodontics, School of Dentistry, Seoul National University) ;
  • Lee, Dong-Soo (Dept. of Nuclear Medicine, College of Medicine, Seoul National University) ;
  • Lee, Seung-Jin (College of Pharmacy, Ewha Womans University) ;
  • Chung, Chong-Pyoung (Dept. of Periodontology, School of Dentistry, Seoul National University) ;
  • Seol, Yang-Jo (Dept. of Periodontology, School of Dentistry, Seoul National University)
  • 김경화 (서울대학교 치과대학 치주과학교실, BK21 치의학생명과학사업단) ;
  • 박윤정 (서울대학교 치과대학 두개악안면재건과학교실) ;
  • 이용무 (서울대학교 치과대학 치주과학교실) ;
  • 한중석 (서울대학교 치과대학 보철학교실) ;
  • 이동수 (서울대학교 의과대학 핵의학과) ;
  • 이승진 (이화여자대학교 약학대학 물리약학교실) ;
  • 정종평 (서울대학교 치과대학 치주과학교실) ;
  • 설양조 (서울대학교 치과대학 치주과학교실)
  • Published : 2006.12.31
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Abstract

The aim of this study is to monitor reporter gene expression under osteocalcin gene promoter, using a real-time molecular imaging system, as tool to investigate osteoblast differentiation. The promoter region of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, was inserted in promoterless luciferase reporter vector. Expression of reporter gene was confirmed and relationship between the reporter gene expression and osteoblastic differentiation was evaluated. Gene expression according to osteoblstic differentiation on biomaterials, utilizing a real-time molecular imaging system, was monitored. Luciferase was expressed at the only cells transduced with pGL4/mOGP and the level of expression was statistically higher at cells cultured in mineralization medium than cells in growth medium. CCCD camera detected the luciferase expression and was visible differentiation-dependent intensity of luminescence. The cells produced osteocalcin with time-dependent increment in BMP-2 treated cells and there was difference between BMP-2 treated cells and untreated cells at 14days. There was difference at the level of luciferase expression under pGL4/mOGP between BMP-2 treated cells and untreated cells at 3days. CCCD camera detected the luciferase expression at cells transduced with pGL4/mOGP on Ti disc and was visible differentiation-dependent intensity of luminescence This study shows that 1) expression of luciferase is regulated by the mouse OC promoter, 2) the CCCD detection system is a reliable quantitative gene detection tool for the osteoblast differentiation, 3) the dynamics of mouse OC promoter regulation during osteoblast differentiation is achieved in real time and quantitatively on biomaterial. The present system is a very reliable system for monitoring of osteoblast differentiation in real time and may be used for monitoring the effects of growth factors, drug, cytokines and biomaterials on osteoblast differentiation in animal.

Keywords

References

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