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(American Journal of Pathology. 2006;169:1440-1457.)
© 2006 American Society for Investigative Pathology
DOI: 10.2353/ajpath.2006.060064

Therapeutic Potential of Vasculogenesis and Osteogenesis Promoted by Peripheral Blood CD34-Positive Cells for Functional Bone Healing

Tomoyuki Matsumoto*{dagger}, Atsuhiko Kawamoto*, Ryosuke Kuroda{dagger}, Masakazu Ishikawa*, Yutaka Mifune*{dagger}, Hiroto Iwasaki*, Masahiko Miwa{dagger}, Miki Horii*, Saeko Hayashi*, Akira Oyamada*, Hiromi Nishimura*, Satoshi Murasawa*, Minoru Doita{dagger}, Masahiro Kurosaka{dagger} and Takayuki Asahara*{ddagger}

From Stem Cell Translational Research,* Kobe Institute of Biomedical Research and Innovation/Riken Center for Developmental Biology, Kobe; the Department of Orthopedic Surgery,{dagger} Kobe University Graduate School of Medicine, Kobe; and the Department of Regenerative Medicine Science,{ddagger} Tokai University School of Medicine, Tokai, Japan

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.




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