Therapeutic potential of vasculogenesis and osteogenesis promoted by peripheral blood CD34-positive cells for functional bone healing
Therapeutic potential of vasculogenesis and osteogenesis promoted by peripheral blood CD34-positive cells for functional bone healing
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DOI：
10.2353/ajpath.2006.060064
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发表时间：
2006-10-01
影响因子：
6
通讯作者：
Asahara, Takayuki
中科院分区：
文献类型：
--
作者：
Matsumoto, Tomoyuki;Kawamoto, Atsuhiko;Asahara, Takayuki
骨折愈合失败主要是由于缺乏血管化。成人循环中的CD 34（+）细胞是一种内皮/造血祖细胞富集的细胞群，据报道在体外可分化为成骨细胞;然而，CD 34（+）细胞用于骨折愈合的治疗潜力仍不清楚。因此，我们进行了一系列实验来验证我们的假设，即功能性骨折愈合是由CD 34（+）细胞再生可塑性的血管生成和骨生成支持的。从成人志愿者的总单核细胞中分离的外周血CD 34（+）细胞，通过单细胞逆转录-聚合酶链反应分析，在20个新鲜分离的细胞中的4个中显示骨钙素基因表达。在裸大鼠股骨骨折不愈合后，静脉内移植磷酸盐缓冲盐水、单核细胞或CD 34（+）细胞。在骨折周围部位的逆转录聚合酶链反应和免疫组化染色表明，在第2周，内皮细胞和成骨细胞的人类特异性标志物的分子和组织学表达。通过生物力学、放射学和组织学检查评估，与其他组相比，CD 34（+）细胞移植显著增强了功能性骨愈合。我们的数据表明，循环中的人CD 34（+）细胞具有治疗潜力，可以促进受损骨骼组织中有利于新血管形成和骨生成的环境，从而使骨折完全愈合。
Failures in fracture healing are mainly caused by a lack of vascularization. Adult human circulating CD34(+) cells, an endothelial/hematopoietic progenitor-enriched cell population, have been reported to differentiate into osteoblasts in vitro; however, the therapeutic potential of CD34(+) cells for fracture healing is still unclear. Therefore, we performed a series of experiments to test our hypothesis that functional fracture healing is supported by vasculogenesis and osteogenesis via regenerative plasticity of CD34(+) cells. Peripheral blood CD34(+) cells, isolated from total mononuclear cells of adult human volunteers, showed gene expression of osteocalcin in 4 of 20 freshly isolated cells by single cell reverse transcriptase-polymerase chain reaction analysis. Phosphate-buffered saline, mononuclear cells, or CD34(+) cells were intravenously transplanted after producing nonhealing femoral fractures in nude rats. Reverse transcriptase-polymerase chain reaction and immunohistochemical staining at the peri-fracture site demonstrated molecular and histological expression of human-specific markers for endothelial cells and osteoblasts at week 2. Functional bone healing assessed by biomechanical as well as radiological and histological examinations was significantly enhanced by CD34(+) cell transplantation compared with the other groups. Our data suggest circulating human CD34(+) cells have therapeutic potential to promote an environment conducive to neovascularization and osteogenesis in damaged skeletal tissue, allowing the complete healing of fractures.