Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression.
Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression.
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DOI：
10.1186/1471-2474-7-62
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发表时间：
2006-08-03
影响因子：
2.3
通讯作者：
Yamamoto T
中科院分区：
文献类型：
--
作者：
Kawai M;Bessho K;Maruyama H;Miyazaki J;Yamamoto T
经皮体内电穿孔有望成为一种有效的基因转移方法，促进骨再生使用的BMP-2质粒载体。为了使用这种方法促进增强的骨诱导，我们同时转移BMP-2和BMP-7的cDNA，作为非病毒载体pCAGGS中的插入物。首先，进行体外研究以确认双基因转移后BMP-2和BMP-7的表达。接下来，将单独的BMP-2和BMP-7质粒或两者一起注射到大鼠小腿肌肉中，并在100 V，50 msec下施加经皮电穿孔8次。在培养系统中，BMP-2和BMP-7基因的同时转移导致C2 C12细胞中的ALP活性比单独转移任一基因高得多。在体内，治疗后10天，软X射线分析显示，接受pCAGGS-BMP-2和pCAGGS-BMP-7的肌肉比接受单一基因的肌肉具有更好的不透明性。组织学检查显示接受双基因转移的小腿肌肉中的晚期骨化。BMP-4 mRNA也有表达，RT-PCR显示双基因转染组BMP-4 mRNA表达水平在3天内呈时间依赖性增加。免疫组织化学证实，BMP-4表达细胞驻留在肌纤维之间的基质中。使用体内电穿孔同时转移BMP-2和BMP-7基因比单独转移任一基因诱导更快的骨形成，并且内源性BMP-4的表达增加表明快速骨化与BMP-4的诱导有关。
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. 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. 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 time-dependent manner in the double-gene transfer group. Immunohistochemistry confirmed that BMP-4-expressing cells resided in the matrix between muscle fibers. 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.
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DOI：
10.1006/bbrc.1999.0341
发表时间：
1999-03-16
影响因子：
3.1
作者：
Hino, J;Matsuo, H;Kangawa, K
通讯作者：
Kangawa, K
影响因子：
3.4
作者：
WOLF, H;ROLS, MP;TEISSIE, J
通讯作者：
TEISSIE, J
影响因子：
1.8
作者：
Israel, DI;Nove, J;Wozney, JM
通讯作者：
Wozney, JM
影响因子：
6.2
作者：
Yoshimura, Y;Nomura, S;Takaoka, K
通讯作者：
Takaoka, K
影响因子：
3.5
作者：
Park, J;Gelse, K;Schneider, H
通讯作者：
Schneider, H