Local delivery of granulocyte colony stimulating factor-mobilized CD34-positive progenitor cells using bioscaffold for modality of unhealing bone fracture
Local delivery of granulocyte colony stimulating factor-mobilized CD34-positive progenitor cells using bioscaffold for modality of unhealing bone fracture
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DOI：
10.1634/stemcells.2007-0820
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发表时间：
2008-06-01
期刊：
影响因子：
5.2
通讯作者：
Asahara, Takayuki
中科院分区：
文献类型：
--
作者：
Mifune, Yutaka;Matsumoto, Tomoyuki;Asahara, Takayuki
我们最近报道，静脉内移植成人循环CD 34+细胞，内皮/造血祖细胞富集的细胞群，有助于骨折愈合，通过增强血管生成和骨生成。然而，成年人中CD 34+细胞的缺乏是该方法未来临床应用的关键问题。为了克服这一问题，我们评估了粒细胞集落刺激因子动员的外周血（GM-PB）人CD 34+细胞的体外和体内血管生成和成骨能力。首先，我们在体外证实了GM-PB CD 34+细胞向成骨细胞的分化能力。第二，在未愈合骨折模型中的裸大鼠中进行GM-PB CD 34+细胞在去端胶原支架上的局部移植。骨折和细胞治疗后1周的组织样本的人类白细胞抗原ABC的免疫染色显示，与全身输注相比，局部移植后的上级掺入。第三，比较局部移植10（5）（Hi）、10（4）（Mid）或10（3）（Lo）剂量的GM-PB CD 34+细胞或磷酸盐缓冲盐水（PBS）对骨折愈合的影响。GM-PB CD 34+细胞的外源性血管生成和成骨分化、受体细胞的内源性血管-骨生成增强、骨折部位血流恢复增强以及骨折愈合的放射学和组织学证实仅在Hi和Mid组中观察到，而在Lo和PBS组中未观察到。这些结果有力地表明，局部移植GM-PB CD 34+细胞与去端胶原支架是一个可行的策略，治疗骨折愈合所需的血管生成和骨生成。
We recently reported that i.v. transplantation of adult human circulating CD34+ cells, an endothelial/hematopoietic progenitor-enriched cell population, contributes to fracture healing through the enhancement of vasculogenesis and osteogenesis. However, the scarcity of CD34+ cells in the adult human is a critical issue for the future clinical application of this method. To overcome this issue, we assessed in vitro and in vivo capacity of granulocyte colony-stimulating factor-mobilized peripheral blood (GM-PB) human CD34+ cells for vasculogenesis and osteogenesis. First, we confirmed the differentiation capability of GM-PB CD34+ cells into osteoblasts in vitro. Second, local transplantation of GM-PB CD34+ cells on atelocollagen scaffold was performed in nude rats in a model of unhealing fractures. Immunostaining for human leukocyte antigen-ABC of tissue samples 1 week after fracture and cell therapy showed the superior incorporation after local transplantation compared with systemic infusion. Third, the effects of local transplantation of 10(5) (Hi), 10(4) (Mid), or 10(3) (Lo) doses of GM-PB CD34+ cells or phosphate-buffered saline (PBS) on fracture healing were compared. Extrinsic vasculogenic and osteogenic differentiation of GM-PB CD34+ cells, enhancement of the intrinsic angio-osteogenesis by recipient cells, augmentation of blood flow recovery at the fracture sites, and radiological and histological confirmation of fracture healing were observed only in the Hi and Mid groups but not in the Lo and PBS groups. These results strongly suggest that local transplantation of GM-PB CD34+ cells with atelocollagen scaffold is a feasible strategy for therapeutic vasculogenesis and osteogenesis needed for fracture healing.