Targeted Gene-and-host Progenitor Cell Therapy for Nonunion Bone Fracture Repair
Targeted Gene-and-host Progenitor Cell Therapy for Nonunion Bone Fracture Repair
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DOI：
10.1038/mt.2010.190
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发表时间：
2011-01-01
影响因子：
12.4
通讯作者：
Gazit, Dan
中科院分区：
文献类型：
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作者：
Kimelman-Bleich, Nadav;Pelled, Gadi;Gazit, Dan
骨不连是骨科面临的一个挑战，目前尚无最佳解决方案。体内DNA电穿孔是一种基因递送技术，可以潜在地加速再生过程。我们假设在桡骨骨不连的骨缺损部位电穿孔成骨基因可以诱导骨折修复。在C3 H/HeN小鼠的桡骨中产生不愈合骨折，其中放置胶原海绵。为了将宿主祖细胞（HPC）募集到植入的海绵中，在电穿孔之前将小鼠圈养10天。用骨形态发生蛋白9（BMP-9）质粒、荧光素酶质粒电穿孔小鼠，或注射BMP-9质粒但不电穿孔。体内生物发光成像表明，基因表达定位于缺陷部位。微计算机断层扫描（μ CT）和组织学分析的小鼠桡骨电穿孔与BMP-9证明骨形成桥接骨间隙，而在对照组中的缺陷保持未桥接。记录电穿孔后用注射的质粒转染的HPC植入的胶原海绵的群体。我们的数据表明，骨不连的骨缺损的再生可以通过在体内进行成骨基因结合募集HPC的电穿孔来实现。这种基因治疗方法可能为其他骨骼组织的再生铺平道路。
Nonunion fractures present a challenge to orthopedics with no optimal solution. In-vivo DNA electroporation is a gene-delivery technique that can potentially accelerate regenerative processes. We hypothesized that in vivo electroporation of an osteogenic gene in a nonunion radius bone defect site would induce fracture repair. Nonunion fracture was created in the radii of C3H/HeN mice, into which a collagen sponge was placed. To allow for recruitment of host progenitor cells (HPCs) into the implanted sponge, the mice were housed for 10 days before electroporation. Mice were electroporated with either bone morphogenetic protein 9 (BMP-9) plasmid, Luciferase plasmid or injected with BMP-9 plasmid but not electroporated. In vivo bioluminescent imaging indicated that gene expression was localized to the defect site. Microcomputed tomography (mu CT) and histological analysis of murine radii electroporated with BMP-9 demonstrated bone formation bridging the bone gap, whereas in the control groups the defect remained unbridged. Population of the implanted collagen sponge by HPCs transfected with the injected plasmid following electroporation was noted. Our data indicate that regeneration of nonunion bone defect can be attained by performing in vivo electroporation with an osteogenic gene combined with recruitment of HPCs. This gene therapy approach may pave the way for regeneration of other skeletal tissues.