Bone regeneration by regulated in vivo gene transfer using Biocompatible polyplex nanomicelles
Bone regeneration by regulated in vivo gene transfer using Biocompatible polyplex nanomicelles
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DOI：
10.1038/sj.mt.6300218
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发表时间：
2007-09-01
影响因子：
12.4
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
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作者：
Itaka, Keiji;Ohba, Shinsuke;Kataoka, Kazunori
基因治疗是骨再生医学的一个很有前途的策略。虽然病毒载体已被深入研究用于递送成骨因子，但免疫应答不可避免地抑制骨形成。因此，安全和有效的非病毒基因递送系统的需求很高。为此，我们开发了由聚乙二醇（PEG）-嵌段共聚物（PEG-b-P[Asp-（DET）]）和质粒DNA（pDNA）组成的复合物纳米胶束体系。该系统对原代细胞的细胞毒性小，转染效率高。通过转染组成型活性形式的激活素受体样激酶6（caALK 6）和runt相关转录因子2（Runx 2），在小鼠颅骨细胞上诱导成骨分化的程度比使用聚乙烯亚胺（PEI）或FuGENE 6时更大;这一结果是由于低细胞毒性和持续的基因表达谱。在掺入磷酸钙骨水泥支架后，复合物纳米胶束成功地从支架释放并转染周围细胞。最后，将该系统应用于小鼠颅骨骨缺损模型的体内基因转移。通过将来自纳米胶束的caALK 6和Runx 2基因引入支架中，诱导了覆盖植入物整个下表面的大量骨形成，在4周时没有炎症迹象。这些结果证明了使用复合物纳米胶束在具有治疗潜力的体内基因转移中的首次成功。
Gene therapy is a promising strategy for bone regenerative medicine. Although viral vectors have been intensively studied for delivery of osteogenic factors, the immune response inevitably inhibits bone formation. Thus, safe and efficient non-viral gene delivery systems are in high demand. Toward this end, we developed a polyplex nanomicelle system composed of poly( ethyleneglycol) ( PEG)-block-catiomer(PEG-b-P[Asp-(DET)]) and plasmid DNA ( pDNA). This system showed little cytotoxicity and excellent transfection efficiency to primary cells. By the transfection of constitutively active form of activin receptor-like kinase 6 ( caALK6) and runt-related transcription factor 2 ( Runx2), the osteogenic differentiation was induced on mouse calvarial cells to a greater extent than when poly( ethylenimine) ( PEI) or FuGENE6 were used; this result was due to low cytotoxicity and a sustained gene expression profile. After incorporation into the calcium phosphate cement scaffold, the polyplex nanomicelles were successfully released from the scaffold and transfected surrounding cells. Finally, this system was applied to in vivo gene transfer for a bone defect model in a mouse skull bone. By delivering caALK6 and Runx2 genes from nanomicelles incorporated into the scaffold, substantial bone formation covering the entire lower surface of the implant was induced with no sign of inflammation at 4 weeks. These results demonstrate the first success in in vivo gene transfer with therapeutic potential using polyplex nanomicelles.