title: Immature muscular tissue differentiation into bone-like tissue by bone morphogenetic proteins in vitro, with ossification potential in vivo.
abstract: The objective of this study was to induce bone formation from immature muscular tissue (IMT) in vitro, using bone morphogenetic proteins (BMPs) as a cytokine source and an expanded polytetrafluoroethylene (ePTFE) scaffold. In addition, cultured IMTs were implanted subcutaneously into Sprague-Dawley (SD) rats to determine their in vivo ossification potential. BMPs, extracted from bovine cortical bones, were applied to embryonic SD rat IMT cultures, before 2 weeks culture on ePTFE scaffolds. Osteoblast-like cells and osteoid tissues were partially identified by hematoxylin-eosin staining 2 weeks after culture. Collagen type I (Col-I), osteopontin (OP), and osteocalcin (OC) were detected in the osteoid tissues by immunohistochemical staining. OC gene expression remained low, but OP and Col-I were upregulated during the culture period. In vivo implanted IMTs showed slight radiopacity 1 week after implantation and strong radiopacity 2 and 3 weeks after implantation. One week after implantation, migration of numerous capillaries was observed and ossification was detected after 2 weeks by histological observation. These results suggest that IMTs are able to differentiate into bone-like tissue in vitro, with an ossification potential after implantation in vivo.
authors.affiliation: Department of Dental Materials Science, Aichi Gakuin University School of Dentistry, Nagoya, Japan.
authors.forename: Tatsuhide
authors.initials: T
authors.lastname: Hayashi
authors.forename: Syuichiro
authors.initials: S
authors.lastname: Kobayashi
authors.forename: Masaki
authors.initials: M
authors.lastname: Asakura
authors.forename: Mayu
authors.initials: M
authors.lastname: Kawase
authors.forename: Atsuko
authors.initials: A
authors.lastname: Ueno
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Uematsu
authors.forename: Tatsushi
authors.initials: T
authors.lastname: Kawai
mesh_terms: D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D008297:Male; D009132:Muscles / Q000166:cytology / Q000187:drug effects* / Q000637:transplantation*; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis / Q000187:drug effects*; D011138:Polytetrafluoroethylene / Q000737:chemistry; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D019485:Bone Morphogenetic Proteins; D011138:Polytetrafluoroethylene
keywords: bone morphogenetic proteins; ePTFE; immature muscular tissue; induced bone-like tissue; ossification potential