Transfer of the bone morphogenetic protein 4 gene into rat periodontal ligament by in vivo electroporation. (February 2017)

• Record Type: Journal Article
• Authors: Tsuchiya, Shinobu; Chiba, Mirei; Kishimoto, Koshi N.; Nakamura, Harukazu; Tsuchiya, Masahiro; Hayashi, Haruhide
• Is Part Of: Archives of oral biology. Volume 74(2017)
• Page Start: 123
• Page End: 132
• Publication Date: 2017-02
• Subjects: ALP alkaline phosphatase -- BMD bone mineral density -- BMP bone morphogenetic protein -- GFP green fluorescent protein -- PBS phosphate-buffered saline -- PDL periodontal ligament; BMP-4 -- Gene transfer -- In vivo electroporation -- Periodontal ligament; Mouth -- Periodicals; Mouth -- Diseases -- Periodicals; Dentistry -- Periodicals; Electronic journals; 617.6005
• DOI: 10.1016/j.archoralbio.2016.11.013
• Languages: English
• ISSNs: 0003-9969

Abstract

Highlights: We established gene transfer into the PDL by electroporation with a plasmid vector. BMP-4 was expressed in PDL cells electroporated with pCAGGS-BMP4 in vitro . BMP-4 was expressed in the rat PDL electroporated with pCAGGS-BMP4 in vivo . The BMP-4 expression level decreased over time and disappeared by day 21. 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. Design: 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 28 days 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.