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TRANSLATIONAL AND CLINICAL RESEARCH |
aDepartment of Orthopedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan;
bStem Cell Translational Research, Kobe Institute of Biomedical Research and Innovation, RIKEN Center for Developmental Biology, Kobe, Japan;
cDepartment of Regenerative Medicine Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan
Key Words. Endothelial progenitor cells • Vascularization • Ligament healing
Correspondence: Correspondence: Ryosuke Kuroda, M.D., Ph.D., Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine 7-5-1, Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan. Telephone: 81-78-382-5985; Fax: 81-78-351-6930; e-mail: kurodar{at}med.kobe-u.ac.jp
Received on August 23, 2007;
accepted for publication on December 25, 2007.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS January 10, 2008.
Neoangiogenesis is a key process in the initial phase of ligament healing. Adult human circulating CD34+ cells, an endothelial/hematopoietic progenitor-enriched cell population, have been reported to contribute to neoangiogenesis; however, the therapeutic potential of CD34+ cells for ligament healing is still unclear. Therefore, we performed a series of experiments to test our hypothesis that ligament healing is supported by CD34+ cells via vasculogenesis. Granulocyte colony-stimulating factor-mobilized peripheral blood (GM-PB) CD34+ cells with atelocollagen (CD34+ group), GM-PB mononuclear cells (MNCs) with atelocollagen (MNC group), or atelocollagen alone (control group) was locally transplanted after the creation of medial collateral ligament injury in immunodeficient rats. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining at the injury site demonstrated that molecular and histological expression of human-specific markers for endothelial cells was higher in the CD34+ group compared with the other groups at week 1. Endogenous effect, assessed by capillary density and mRNA expression of vascular endothelial growth factor, was significantly higher in CD34+ cell group than the other groups. In addition to the observation that, as assessed by real-time RT-PCR, gene expression of ligament-specific marker was significantly higher in the CD34+ group than in the other groups, ligament healing assessed by macroscopic, histological, and biomechanical examination was significantly enhanced by CD34+ cell transplantation compared with the other groups. Our data strongly suggest that local transplantation of circulating human CD34+ cells may augment the ligament healing process by promoting a favorable environment through neovascularization.
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