title: The effect of simulated microgravity by three-dimensional clinostat on bone tissue engineering.
abstract: Evidence suggests that mechanical stress, including gravity, is associated with osteoblast differentiation and function. To examine effects of microgravity on bone tissue engineering, we used a three-dimensional (3D) clinostat manufactured by Mitsubishi Heavy Industries (Kobe, Japan). A 3D clinostat is a device that generates multidirectional G force. By controlled rotation on two axes, it cancels the cumulative gravity vector at the center of the device. We cultured rat marrow mesenchymal cells (MMCs) in the pores of interconnected porous calcium hydroxyapatite (IP-CHA) for 2 weeks in the presence of dexamethasone using the 3D clinostat (clinostat group). MMCs cultured using the 3D clinostat exhibited a 40% decrease in alkaline phosphatase activity (a marker of osteoblastic differentiation), compared with control static cultures (control group). SEM analysis revealed that although there was no difference between the two groups in number or distribution of cells in the pores, the clinostat group exhibited less extensive extracellular matrix formation than the control group. Cultured IP-CHA/MMC composites were then implanted into subcutaneous sites of syngeneic rats and harvested 8 weeks after implantation. All implants showed bone formation inside the pores, as indicated by decalcified histological sections and microfocus computed tomography. However, the volume of newly formed bone was significantly lower for the clinostat group than for the control group, especially in the superficial pores close to the implant surface. These results indicate that new bone formation in culture was inhibited by use of the 3D clinostat, and that this inhibition was mainly due to suppression of osteoblastic differentiation of MMCs.
authors.affiliation: Department of Orthopaedics, Osaka University Graduate School of Medicine, Suita City, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Nishikawa
authors.forename: Hajime
authors.initials: H
authors.lastname: Ohgushi
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Tamai
authors.forename: Koichi
authors.initials: K
authors.lastname: Osuga
authors.forename: Masaru
authors.initials: M
authors.lastname: Uemura
authors.forename: Hideki
authors.initials: H
authors.lastname: Yoshikawa
authors.forename: Akira
authors.initials: A
authors.lastname: Myoui
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D001842:Bone and Bones / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D017886:Durapatite; D005109:Extracellular Matrix / Q000502:physiology; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D010012:Osteogenesis; D051381:Rats; D011916:Rats, Inbred F344; D023822:Tissue Engineering / Q000379:methods*; D018474:Weightlessness Simulation / Q000295:instrumentation
chemical_list: D003907:Dexamethasone; D017886:Durapatite