Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression

Kawai, Mariko; Bessho, Kazuhisa; Maruyama, Hiroki; Miyazaki, Jun-ichi; Yamamoto, Toshio

doi:https://doi.org/10.1186/1471-2474-7-62

アクセス数:286件（2024-12-04 01:58 集計）

固定URL: https://hdl.handle.net/11094/23110

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タイトル	Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression
著者	NRID 1000040379839 1000040379839 著者名 Kawai, Mariko
	NRID 1000090229138 1000090229138 著者名 Bessho, Kazuhisa
	NRID 1000010293218 1000010293218 著者名 Maruyama, Hiroki
	NRID 1000080229830 1000080229830 著者名 Miyazaki, Jun-ichi
	NRID 1000030107776 1000030107776 著者名 Yamamoto, Toshio
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.
出版者	BioMed Central
収録物名	BMC Musculoskeletal Disorders
巻(号)	7(62)
ページ	1-11
刊行年月	2006-08-03
言語	英語
ハンドルURL	https://hdl.handle.net/11094/23110
eISSN	14712474
NCID	AA12035278
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1186/1471-2474-7-62
アクセス権	open access
権利情報	© 2006 Kawai et al; licensee BioMed Central Ltd. This article is licensed under a Creative Commons Attribution 2.0 Generic License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	BMC Musculoskeletal Disorders. 2006, 7(62), p. 1-11
DC.title	Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression
DC.creator	Kawai, Mariko
	Bessho, Kazuhisa
	Maruyama, Hiroki
	Miyazaki, Jun-ichi
	Yamamoto, Toshio
DC.publisher	BioMed Central
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2006-08-03
DC.identifier	https://doi.org/10.1186/1471-2474-7-62
DCTERMS.abstract	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.
DC.rights	© 2006 Kawai et al; licensee BioMed Central Ltd. This article is licensed under a Creative Commons Attribution 2.0 Generic License.
DC.format	23110
citation_title	Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression
citation_author	Kawai, Mariko
	Bessho, Kazuhisa
	Maruyama, Hiroki
	Miyazaki, Jun-ichi
	Yamamoto, Toshio
citation_publisher	BioMed Central
citation_language	英語
citation_date	2006-08-03
citation_journal_title	BMC Musculoskeletal Disorders
citation_volume	7
citation_issue	62
citation_firstpage	1
citation_lastpage	11
citation_doi	https://doi.org/10.1186/1471-2474-7-62
citation_public_url	https://hdl.handle.net/11094/23110

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