DNA nanoparticles encapsulated in 3D tissue-engineered scaffolds enhance osteogenic differentiation of mesenchymal stem cells
DNA nanoparticles encapsulated in 3D tissue-engineered scaffolds enhance osteogenic differentiation of mesenchymal stem cells
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DOI：
10.1002/jbm.a.31327
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发表时间：
2008-04-01
影响因子：
4.9
通讯作者：
Kobayashi, Hisatoshi
中科院分区：
文献类型：
--
作者：
Hosseinkhani, Hossein;Hosseinkhani, Mohsen;Kobayashi, Hisatoshi
在这项研究中，我们通过三维（3D）组织工程支架和非病毒基因载体的结合来增强质粒DNA在间充质干细胞（MSC）中的表达。为了增强质粒DNA的传递功能，乙酸酐与聚乙烯亚胺（PEI）反应，使80%的一级胺和20%的二级胺乙酰化（PEI- ac(80)）。这种乙酰化的PEI已被证明比未修饰的PEI表现出更高的基因传递效率。采用聚乙醇酸（PGA）纤维增强的胶原海绵作为支架材料。编码骨形态发生蛋白-2 （BMP-2）的质粒DNA与PEI-Ac-80溶液简单混合形成的DNA纳米颗粒被封装在这些支架内。将MSC植入每个支架并培养数周。在这些支架中，与DNA纳米颗粒溶液（在二维组织培养板中）转染的间充质干细胞相比，转染的间充质干细胞的BMP-2表达水平显著提高。在大鼠背部皮下植入DNA纳米颗粒转染骨髓间充质干细胞后，在组织学上观察到整个海绵均有均匀的骨形成。转染骨髓间充质干细胞后，海绵植入部位的碱性磷酸酶活性和骨钙素含量明显高于其他基质。(c) 2007 Wiley期刊公司
In this study, we enhanced the expression of a plasmid DNA in mesenchymal stem cells (MSC) by the combination of three-dimensional (3D) tissue-engineered scaffold and nonviral gene carrier. To function as an enhanced delivery of plasmid DNA, acetic anhydride was reacted with polyethylenimine (PEI) to acetylate 80% of the primary and 20% of the secondary amines (PEI-AC(80)). This acetylated PEI has been demonstrated to show enhanced gene-delivery efficiency over unmodified PEI. Collagen sponges reinforced by incorporating of poly(glycolic acid) (PGA) fibers were used as the scaffold material. DNA nanoparticles formed through simple mixing of plasmid DNA encoding bone morphogenctic protein-2 (BMP-2) and PEI-Ac-80 solutions were encapsulated within these scaffolds. MSC were seeded into each scaffold and cultured for several weeks. Within these scaffolds, the level of BMP-2 expression by transfected MSC was significantly enhanced compared to MSC transfected by DNA nanoparticles in solution (in 2D tissue culture plates). Homogeneous bone formation was histologically observed throughout the sponges seeded with transfected MSC by using DNA nanoparticles after subcutaneous implantation into the back of rats. The level of alkaline phosphatase activity and osteocalcin content at the implanted sites of sponges seeded with transfected MSC by using DNA nanoparticles were significantly higher when compared with those seeded with other agents. (c) 2007 Wiley Periodicals, Inc.