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首页> 外文期刊>Cell biology international. >Ectopic study of calcium phosphate cement seeded with pBMP-2 modified canine bMSCs mediated by a non-viral PEI derivative.
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Ectopic study of calcium phosphate cement seeded with pBMP-2 modified canine bMSCs mediated by a non-viral PEI derivative.

机译：非病毒性PEI衍生物介导的pBMP-2修饰的犬bMSC植入磷酸钙水泥的异位研究。

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摘要

We have evaluated the ectopic new bone formation effects of CPC (calcium phosphate cement) seeded with pBMP-2 (plasmids containing bone morphogenetic protein-2 gene) transfected canine bMSCs (bone marrow stromal cells) mediated by a non-viral PEI (polyethylenimine) derivative (GenEscort? II) in nude mice. Canine bMSCs were transfected with pBMP-2 or pEGFP (plasmids containing enhanced green fluorescent protein gene) mediated by GenEscort? II in vitro, and the osteoblastic differentiation was explored by ALP (alkaline phosphatase) staining, ARS (alizarin red S) staining and RT-qPCR (real-time quantitative PCR) analysis. Ectopic bone formation effects of CPC/pBMP-2 transfected bMSCs were evaluated and compared with CPC/pEGFP transfected bMSCs or CPC/untransfected bMSCs through histological, histomorphological and immunohistochemical analysis 8 and 12 weeks post-operation in nude mice. Transfection efficiency was up ～35% as demonstrated by EGFP (enhanced green fluorescent protein) expression. ALP and ARS staining were stronger with pBMP-2 gene transfection, and mRNA expression of BMP-2 (bone morphogenetic protein-2), Col 1 (collagen 1) and OCN (osteocalcin) in pBMP-2 group was significantly up-regulated at 6 and 9 days. Significantly higher NBV (new bone volume) was achieved in pBMP-2 group than in the control groups at 8 and 12 weeks (P<0.05). In addition, immunohistochemical analysis indicated higher OCN expression in pBMP-2 group (P<0.01). We conclude that CPC seeded with pBMP-2 transfected bMSCs mediated by GenEscort? II could enhance ectopic new bone formation in nude mice, suggesting that GenEscort? II mediated pBMP-2 gene transfer is an effective non-viral method and CPC is a suitable scaffold for gene enhanced bone tissue engineering.
机译：我们已经评估了通过非病毒性PEI（聚乙烯亚胺）介导的pBMP-2（含有骨形态发生蛋白2基因的质粒）转染的犬bMSCs（骨髓基质细胞）接种的CPC（磷酸钙水泥）的异位新骨形成作用衍生物（GenEscort？II）在裸鼠中。用GenEscort介导的pBMP-2或pEGFP（含有增强的绿色荧光蛋白基因的质粒）转染犬bMSCs？体外，通过ALP（碱性磷酸酶）染色，ARS（茜素红S）染色和RT-qPCR（实时定量PCR）分析探索成骨细胞的分化。评价裸鼠术后8、12周通过CPC / pBMP-2转染的bMSCs异位骨形成的影响，并与CPC / pEGFP转染的bMSCs或CPC /未转染的bMSCs进行组织学，组织形态学和免疫组织化学分析。 EGFP（增强型绿色荧光蛋白）表达证明转染效率高达〜35％。 pBMP-2基因转染后ALP和ARS染色更强，pBMP-2组的BMP-2（骨形态发生蛋白2），Col 1（胶原1）和OCN（骨钙蛋白）的mRNA表达在90℃时显着上调。 6和9天。在第8周和第12周，pBMP-2组的NBV（新骨量）明显高于对照组（P <0.05）。此外，免疫组织化学分析表明pBMP-2组的OCN表达较高（P <0.01）。我们得出的结论是，GenEscort介导了用pBMP-2转染的bMSC接种的CPC。 II可以增强裸鼠异位新骨的形成，提示GenEscort？ II介导的pBMP-2基因转移是一种有效的非病毒方法，CPC是基因增强骨组织工程的合适支架。

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