title: Acceleration of bone formation during fracture healing by poly(pro-hyp-gly)10 and basic fibroblast growth factor containing polycystic kidney disease and collagen-binding domains from Clostridium histolyticum collagenase.
abstract: Growth factor delivered in combination with animal-derived collagen materials has been used to accelerate bone fracture healing in human patients. However, the introduction of bovine proteins into humans carries the risk of zoonotic and immunologic complications. Here, we developed a collagen-like polypeptide-based bone formation system consisting of poly(Pro-Hyp-Gly)10 , which mimics the triple helical conformation of collagen, and basic fibroblast growth factor (bFGF) fused to the polycystic kidney disease (PKD) domain and collagen-binding domain (CBD) of Clostridium histolyticum collagenase. Circular dichroism spectral analysis showed that when pepsin-soluble bovine type I collagen was treated at 50°C, a positive signal corresponding to the collagen triple helix at 220 nm was not detected. In contrast, poly(Pro-Hyp-Gly)10 retained the 220-nm positive peak, even when treated at 80°C. The combination of the collagen binding-bFGF fusion protein (bFGF-PKD-CBD) with poly(Pro-Hyp-Gly)10 induced greater bone formation compared to bFGF alone in mice bone fracture models. Taken together, these properties suggest that the bFGF-PKD-CBD/poly(Pro-Hyp-Gly)10 composite is a promising material for bone repair in the clinical setting. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1372-1378, 2016.
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sekiguchi
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Gen
authors.initials: G
authors.lastname: Inoue
authors.affiliation: Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Kita-Ku Shikata-Cho, Okayama, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Matsushita
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Saito
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Aikawa
authors.affiliation: Nippi Research Institute of Biomatrix and Protein Engineering Project, 520-11, Kuwabara Toride-Shi, Ibaraki-Ken, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Hisako
authors.initials: H
authors.lastname: Fujimaki
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Miyagi
authors.affiliation: Department of Orthopedic Surgery, Kitasato University School of Medicine, 1-15-1 Minami-Ku, Kitasato, Sagamihara City, Kanagawa, 252-0374, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Takaso
mesh_terms: D000818:Animals; D002146:Bony Callus / Q000187:drug effects / Q000473:pathology; D002417:Cattle; D002942:Circular Dichroism; D047009:Hathewaya histolytica / Q000201:enzymology*; D017364:Collagenases / Q000737:chemistry*; D005269:Femur / Q000000981:diagnostic imaging / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000494:pharmacology*; D017102:Fracture Healing / Q000187:drug effects*; D006801:Humans; D021621:Imaging, Three-Dimensional; D008810:Mice, Inbred C57BL; D010012:Osteogenesis / Q000187:drug effects*; D010316:Particle Size; D010455:Peptides / Q000494:pharmacology*; D000072417:Protein Domains; D055114:X-Ray Microtomography
chemical_list: D010455:Peptides; D016222:Fibroblast Growth Factor 2; C083881:(prolyl-hydroxylprolyl-glycine)10; D017364:Collagenases
keywords: basic fibroblast growth factor; collagen-binding domain; fracture healing; poly(Pro-Hyp-Gly)10