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ID 32941
フルテキストURL
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著者
Kawai, Mariko Department of Oral Morphology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
Bessho, Kazuhisa Department of Oral and Maxillofacial Surgery, Graduate School of Medicine, Kyoto University
Maruyama, Hiroki Division of Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences
Miyazaki, Jun-ichi Division of Stem Cell Regulation Research, Osaka University Medical School
Yamamoto, Toshio Department of Oral Morphology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
抄録

Background: Transcutaneous in vivo electroporation is expected to be an effective gene-transfer method for promoting bone regeneration using the BMP-2 plasmid vector. To promote enhanced osteoinduction using this method, we simultaneously transferred cDNAs for BMP-2 and BMP-7, as inserts in the non-viral vector pCAGGS.
Methods: First, an in vitro study was carried out to confirm the expression of BMP-2 and BMP-7 following the double-gene transfer. Next, the individual BMP-2 and BMP-7 plasmids or both together were injected into rat calf muscles, and transcutaneous electroporation was applied 8 times at 100 V, 50 msec.
Results: In the culture system, the simultaneous transfer of the BMP-2 and BMP-7 genes led to a much higher ALP activity in C2C12 cells than did the transfer of either gene alone. In vivo, ten days after the treatment, soft X-ray analysis showed that muscles that received both pCAGGS-BMP-2 and pCAGGS-BMP-7 had better-defined opacities than those receiving a single gene. Histological examination showed advanced ossification in calf muscles that received the double-gene transfer. BMP-4 mRNA was also expressed, and RT-PCR showed that its level increased for 3 days in a timedependent manner in the double-gene transfer group. Immunohistochemistry confirmed that BMP- 4-expressing cells resided in the matrix between muscle fibers.
Conclusion: The simultaneous transfer of BMP-2 and BMP-7 genes using in vivo electroporation induces more rapid bone formation than the transfer of either gene alone, and the increased expression of endogenous BMP-4 suggests that the rapid ossification is related to the induction of BMP-4.

備考
Digital Object Identifer:10.1186/1471-2474-7-62
Published with permission from the copyright holder. This is the institute's copy, as published in BMC Musculoskeletal Disorders, 3 August 2006, 7:62.
Publisher URL:http://dx.doi.org/10.1186/1471-2474-7-62
© 2006 Kawai et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
発行日
2006-08-03
出版物タイトル
BMC Musculoskeletal Disorders
7巻
出版者
BioMed Central
NCID
AA12035278
資料タイプ
学術雑誌論文
言語
English
OAI-PMH Set
岡山大学
著作権者
Kawai et al; licensee BioMed Central Ltd.
論文のバージョン
publisher
査読
有り
DOI
PubMed ID
Submission Path
internal_medicine/4