Maxillofacial Plastic and Reconstructive Surgery

Korean Association of Maxillofacial Plastic and Reconstructive Surgeons (대한악안면성형재건외과학회)
권호 표지

TOPICAL GENE DELIVERY TO NORMAL ORAL EPITHELIUM USING ADENOVIRUS IN ORGAN CULTURE MODEL

조직 배양 모형에서 정상 구강 점막 상피에 대한 국소 유전자 요법

  • Kim, Tae-Hwan (School of Dentistry, College of Medicine, Hallym University) ;
  • Kwak, Myung-Ho (School of Dentistry, College of Medicine, Hallym University) ;
  • Lee, Choon-Ho (School of Dentistry, College of Medicine, Hallym University) ;
  • Park, Jun-Woo (School of Dentistry, College of Medicine, Hallym University) ;
  • Park, Young-Wook (Dept. of Oral and Maxillofacial Surgery, College of Dentistry, Kangnung-Wonju National University) ;
  • Kim, Seong-Gon (School of Dentistry, College of Medicine, Hallym University)
  • 김태환 (한림대학교 의과대학 치과학교실) ;
  • 곽명호 (한림대학교 의과대학 치과학교실) ;
  • 이춘호 (한림대학교 의과대학 치과학교실) ;
  • 박준우 (한림대학교 의과대학 치과학교실) ;
  • 박영욱 (강릉원주대학교 치과대학 구강악안면외과학교실) ;
  • 김성곤 (한림대학교 의과대학 치과학교실)
  • Received : 2009.03.26
  • Accepted : 2009.05.19
  • Published : 2009.05.30
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Abstract

Background: Though it is clear that many types of viruses can infect the oral mucosa, its condition for infection is unclear. The purpose of this study was to analyze the conditions for viral infection of normal oral mucosa and explore the possibility of topical gene therapy to oral mucosa using a viral vector. Methods: Freshly taken fragments of the palate and the tongue of mice were used for organ culture. The specimens were exposed to green fluorescent protein (GFP)-adenoviral vector for 1 hour except for the control. Initial viral titer was $6.3{\times}10^{11}\;pfu/ml$ and the virus was diluted to working concentrations. The dilution ratio was 1:1,000 ($6.3{\times}10^8\;pfu/ml$), 1:10,000 ($6.3{\times}10^7\;pfu/ml$), and 1:100,000 ($6.3{\times}10^6\;pfu/ml$). They were then cultured on a stainless steel wire mesh in an organ culture dish. The specimens were stereoscopically examined every 24 hours for 6 days, after which they were fixed and analyzed through immunohistochemical methods Results: There was no visible expression in the control, $6.3{\times}10^6\;pfu/ml$, and $6.3{\times}10^7\;pfu/ml$ groups. Initial expression was observed at 24 hours after infection in both the palate and the tongue in $6.3{\times}10^8\;pfu/ml$ and the expression significantly increased until 3 days in the palate and 2 days in the tongue after infection (P<0.05). In both groups, the expression was mostly observed at the resection margin. Immunohistochemical studies showed that the epithelial cells were positive to GFP. Conclusion: The present study showed that topically applied adenovirus containing specific genetic information of GFP could successfully transduce GFP in normal oral epithelial cells at the resection margin in organ culture in terms of dose and exposure time.

Keywords

References

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