Efficiently engineered cell sheet using a complex of polyethylenimine-alginate nanocomposites plus bone morphogenetic protein 2 gene to promote new bone formation.
Efficiently engineered cell sheet using a complex of polyethylenimine-alginate nanocomposites plus bone morphogenetic protein 2 gene to promote new bone formation.
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DOI：
10.2147/ijn.s60937
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发表时间：
2014
影响因子：
8
通讯作者：
Sun H
中科院分区：
文献类型：
--
作者：
Jin H;Zhang K;Qiao C;Yuan A;Li D;Zhao L;Shi C;Xu X;Ni S;Zheng C;Liu X;Yang B;Sun H
大面积骨缺损的修复是临床上常见的问题。近年来，干细胞片已成为骨组织工程的一种新兴策略。为了提高干细胞片的成骨能力，我们采用聚乙烯亚胺-海藻酸钠（PEI-al）纳米复合材料与人骨形态发生蛋白2（BMP-2）互补（c）DNA质粒复合的方法构建了BMP-2基因工程细胞片，并研究了其在体外和体内的成骨能力。携带BMP-2基因的PEI-Al纳米复合材料可有效地转染骨髓间充质干细胞。通过在含有维生素C的培养基中培养细胞10天来制备细胞片。对细胞培养物的分析表明，基因工程细胞释放BMP-2至少14天。成骨相关基因的表达增加，表明释放的BMP-2能有效诱导细胞片体外成骨分化。为了进一步测试细胞片在体内的成骨潜力，在颅骨缺损上处理增强的绿色荧光蛋白或BMP-2产生细胞片。结果表明，BMP-2产生细胞片组在促进缺损区骨形成方面比其他组更有效。我们的研究结果表明，PEI-Al纳米复合材料有效地将BMP-2基因传递到骨髓间充质干细胞，BMP-2基因工程细胞片是促进骨再生的有效方法。
Regeneration of large bone defects is a common clinical problem. Recently, stem cell sheet has been an emerging strategy in bone tissue engineering. To enhance the osteogenic potential of stem cell sheet, we fabricated bone morphogenetic protein 2 (BMP-2) gene-engineered cell sheet using a complex of polyethylenimine–alginate (PEI–al) nanocomposites plus human BMP-2 complementary(c)DNA plasmid, and studied its osteogenesis in vitro and in vivo. PEI–al nanocomposites carrying BMP-2 gene could efficiently transfect bone marrow mesenchymal stem cells. The cell sheet was made by culturing the cells in medium containing vitamin C for 10 days. Assays on the cell culture showed that the genetically engineered cells released the BMP-2 for at least 14 days. The expression of osteogenesis-related gene was increased, which demonstrated that released BMP-2 could effectively induce the cell sheet osteogenic differentiation in vitro. To further test the osteogenic potential of the cell sheet in vivo, enhanced green fluorescent protein or BMP-2-producing cell sheets were treated on the cranial bone defects. The results indicated that the BMP-2-producing cell sheet group was more efficient than other groups in promoting bone formation in the defect area. Our results suggested that PEI–al nanocomposites efficiently deliver the BMP-2 gene to bone marrow mesenchymal stem cells and that BMP-2 gene-engineered cell sheet is an effective way for promoting bone regeneration.