title: Local transplantation of granulocyte colony-stimulating factor-mobilized human peripheral blood mononuclear cells for unhealing bone fractures.
abstract: We previously reported the therapeutic potential of human peripheral blood (hPB) CD34(+) cells for bone fracture healing via vasculogenesis/angiogenesis and osteogenesis. Transplantation of not only hPB CD34(+) cells but also hPB total mononuclear cells (MNCs) has  shown their therapeutic efficiency for enhancing ischemic neovascularization. Compared with transplantation of purified hPB CD34(+) cells, transplantation of hPB MNCs is more attractive due to its simple method of cell isolation and inexpensive cost performance in the clinical setting.  Thus, in this report, we attempted to test a hypothesis that granulocyte colony-stimulating factor-mobilized (GM) hPB MNC transplantation could also contribute to fracture healing via vasculogenesis/angiogenesis and osteogenesis. Nude rats with unhealing fractures received local administration  of the following materials with atelocollagen: 1 × 10(7) GM hPB MNCs (Hi group), 1 × 10(6) GM hPB MNCs (Lo group), or PBS (PBS group). Immunohistochemistry and real-time reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrated human cell-derived vasculogenesis  and osteogenesis in the Hi and Lo groups, but not in the PBS group at week 1. Intrinsic angiogenesis and osteogenesis assessed by rat capillary, osteoblast density, and real-time RT-PCR analysis was significantly enhanced in the Hi group compared to the other groups. Blood flow assessment  by laser doppler perfusion imaging showed a significantly higher blood flow ratio at week 1 in the Hi group compared with the other groups. Morphological fracture healing was radiographically and histologically confirmed in about 30% of animals in the Hi group at week 8, whereas all animals  in the other groups resulted in nonunion. Local transplantation of GM hPB MNCs contributes to fracture healing via vasculogenesis/angiogenesis and osteogenesis.
authors.affiliation: Group of Vascular Regeneration, Institute of Biomedical Research and Innovation, Kobe, Hyogo, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Fukui
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Mifune
authors.forename: Taro
authors.initials: T
authors.lastname: Shoji
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kuroda
authors.forename: Yohei
authors.initials: Y
authors.lastname: Kawakami
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Kawamoto
authors.forename: Masaaki
authors.initials: M
authors.lastname: Ii
authors.forename: Shin
authors.initials: S
authors.lastname: Kawamata
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kurosaka
authors.forename: Takayuki
authors.initials: T
authors.lastname: Asahara
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Kuroda
mesh_terms: D000328:Adult; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D050723:Fractures, Bone / Q000628:therapy*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D007963:Leukocytes, Mononuclear / Q000187:drug effects*; D008297:Male; D018919:Neovascularization, Physiologic / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology; D051381:Rats; D011923:Rats, Nude; D060888:Real-Time Polymerase Chain Reaction; D055815:Young Adult
chemical_list: D018952:Antigens, CD34; D016179:Granulocyte Colony-Stimulating Factor