title: Transfer of the bone morphogenetic protein 4 gene into rat periodontal ligament by in vivo electroporation.
abstract: OBJECTIVE
  Regulation of alveolar bone metabolism is required in clinical dentistry. The aim of the present study was to establish a method for gene transfer into the periodontal ligament (PDL) by in vivo electroporation with a plasmid vector and to investigate the effects of BMP-4 transfer into the PDL.
  
  
  METHODS
  Plasmids containing mouse BMP-4 cDNA (pCAGGS-BMP4) were transfected into cultured rat PDL cells by in vitro electroporation, and BMP-4 production and secretion were detected by immunocytochemistry and western blotting. Next, pCAGGS-BMP4 was injected into the PDL of rats, and electroporation was performed in vivo, using original paired-needle electrodes. BMP-4 expression was examined by immunohistochemical staining 3, 7, 14, 21, and 28days after electroporation. Control groups were injected with pCAGGS by electroporation, injected with pCAGGS-BMP4 without electroporation, or subjected to neither injection nor electroporation.
  
  
  RESULTS
  In vitro-transfected rat PDL cells exhibited production and secretion of the mature-form BMP-4. After in vivo electroporation of pCAGGS-BMP4, site-specific BMP-4 expression peaked on day 3, gradually decreased until day 14, and was absent by day 21. We observed no unfavorable effects such as inflammation, degeneration, or necrosis.
  
  
  CONCLUSIONS
  Gene transfer by electroporation with plasmid DNA vectors has several advantages over other methods, including the non-viral vector, non-immunogenic effects, site-specific expression, simplicity, cost-effectiveness, and limited histological side effects. Our results indicate that the method is useful for gene therapy targeting the periodontal tissue, which regulates alveolar bone remodeling.
authors.affiliation: Division of Oral Dysfunction Science, Department of Oral Health and Development Sciences, Tohoku University Graduate School of Dentistry, Miyagi, 980-8575, Japan. Electronic address: shinobu.tsuchiya.c2@tohoku.ac.jp.
authors.forename: Shinobu
authors.initials: S
authors.lastname: Tsuchiya
authors.affiliation: Division of Oral Physiology, Department of Oral Function and Morphology, Tohoku University Graduate School of Dentistry, Miyagi, 980-8575, Japan. Electronic address: mirei@m.tohoku.ac.jp.
authors.forename: Mirei
authors.initials: M
authors.lastname: Chiba
authors.affiliation: Department of Orthopaedic Surgery, Tohoku University Graduate School of Medicine, Miyagi, 980-8575, Japan. Electronic address: kishimoto@med.tohoku.ac.jp.
authors.forename: Koshi N
authors.initials: KN
authors.lastname: Kishimoto
authors.affiliation: Department of Molecular Neurobiology, Tohoku University Graduate School of Life Sciences and Institute of Development, Aging and Cancer, Miyagi, 980-8575, Japan. Electronic address: harukazunakamura@gmail.com.
authors.forename: Harukazu
authors.initials: H
authors.lastname: Nakamura
authors.affiliation: Faculty of Health Science, Department of Nursing, Tohoku Fukushi University, Miyagi, 981-8522, Japan; Division of Oral Diagnosis, Tohoku University Graduate School of Dentistry, Miyagi, 980-8575, Japan. Electronic address: mtsuchiya@tfu-mail.tfu.ac.jp.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Tsuchiya
authors.affiliation: Division of Oral Physiology, Department of Oral Function and Morphology, Tohoku University Graduate School of Dentistry, Miyagi, 980-8575, Japan. Electronic address: hayashi@m.tohoku.ac.jp.
authors.forename: Haruhide
authors.initials: H
authors.lastname: Hayashi
mesh_terms: D000818:Animals; D015519:Bone Density; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics* / Q000378:metabolism*; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007150:Immunohistochemistry; D007267:Injections / Q000295:instrumentation / Q000379:methods; D008297:Male; D051379:Mice; D010513:Periodontal Ligament / Q000166:cytology / Q000000981:diagnostic imaging / Q000378:metabolism*; D010957:Plasmids; D011859:Radiography; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods
chemical_list: D055415:Bone Morphogenetic Protein 4
keywords: BMP-4; Gene transfer; In vivo electroporation; Periodontal ligament