转基因骨髓间质干细胞在组织工程承载体中生长及成骨转化的研究

[目的]观察经携带人骨形态发生蛋白-2(BMP-2)基因的复制缺陷重组腺病毒(Ad-BMP-2)转染的兔骨髓间质干细胞(MSCs)在以兔脱细胞脱钙骨基质作为组织工程承载体中的生长情况及转基因前后细胞成骨能力的变化。[方法]用携带有人BMP-2基因片段的复制缺陷重组腺病毒(Ad-BMP-2)转染兔骨髓间质干细胞并将其种植在兔脱细胞脱钙骨基质支架中,扫描电镜观察转基因细胞在支架中的黏附生长情况;并通过检测培养上清中的碱性磷酸酶(ALP)、骨钙素(BGP)含量及Ⅰ型胶原(CollagenⅠ)的分泌量来评估转基因对MSCs成骨能力的影响。[结果]转基因骨髓间质干细胞在组织工程支架中黏附生长良好,转基因组细胞的ALP、BGP含量及Ⅰ型胶原的分泌量与对照组比较差异有显著性意义。[结论]转基因骨髓间质干细胞在兔脱细胞脱钙骨基质支架中能很好的黏附并立体生长,转基因细胞在目的基因所表达的BMP-2蛋白作用下成骨能力明显增强。

Study on the growth status and osteogenic potential of gene transferred mesenchymal stem cells with Ad-BMP-2 planted on tissue engineering scaffold

[Objective]To observe the growth status and osteogenic potential of gene transferred mesenchymal stem cells with Ad-BMP-2 planted on allogenetic acellular and decalcified bone matrix served as tissue engineering scaffold.[Method]Rabbit mesanchymal stem cells transferred with Ad-BMP-2 were seeded on allogenetic acellular and decalcified bone matrix,the growth status of gene transferred MSCs on the scaffold was observed by scanning electron microscopy and the ostengenic potential was observed by testing contents of ALP and BGP and secretion of Collagen I in the culture supermatant.[Result]The MSCs transfected with Ad-BMP-2 grew well on the tissue engineering scaffold,the osteogenic ability of the gene transfected MSCs showed great significant difference compared with that of the control group.[Conclusion]The MSCs transfected with Ad-BMP-2 can grow well on the acellular and decalcified bone matrix and the osteogenic potential can be great improved by gene transfection.