title: Synergistic effect of ascorbic acid and collagen addition on the increase in type 2 collagen accumulation in cartilage-like MSC sheet.
abstract: Aiming to increase the content of type 2 collagen in scaffold-free cartilage-like cell sheets prepared using human bone marrow mesenchymal stem cells, the effect of several kinds of additives in a chondrogenic medium was investigated. Addition of ascorbic acid 2 phosphate (VCP) at a high concentration (250 µg/ml) and type 1 atelocollagen (5 µg/ml) increased the accumulation of type 2 collagen by fourfold and twofold, respectively. On the other hand, an antioxidant, glutathione showed no such effect. The synergistic effect of VCP and type 1 atelocollagen resulted in an eightfold increase in the accumulation level of type 2 collagen. Furthermore, the gene expression level of type 2 collagen increased and that of matrix metalloproteinase-13 (MMP-13) decreased to approximately one-third of the control. The increase in type 2 collagen accumulation in the scaffold-free cartilage-like cell sheet might be due to not only the enhancement of the synthesis but also the suppression of the degradation of type 2 collagen by MMP-13.
authors.affiliation: Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Kita-ku, N13W8, Sapporo, 060-8628, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Sato
authors.affiliation: School of Health and Sports Sciences, Mukogawa Women's University, 6-46 Ikebiraki, Nishinomiya, Hyogo, 663-8558, Japan.|Foundation for Biomedical Research and Innovation, International Medical Device Alliance, 1-6-5, Minatojima Minamimachi, Chuo-ku, Kobe, Hyogo, 650-0047, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Mera
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine, Hokkaido University, Kita-ku, N15W7, Sapporo, 060-8638, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Takahashi
authors.affiliation: Department of Joint Replacement and Tissue Engineering, Graduate School of Medicine, Hokkaido University, Kita-ku, N15W7, Sapporo, 060-8638, Japan.
authors.forename: Tokifumi
authors.initials: T
authors.lastname: Majima
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine, Hokkaido University, Kita-ku, N15W7, Sapporo, 060-8638, Japan.
authors.forename: Norimasa
authors.initials: N
authors.lastname: Iwasaki
authors.affiliation: Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 739-8553, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Wakitani
authors.affiliation: Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Kita-ku, N13W8, Sapporo, 060-8628, Japan. takagi-m@eng.hokudai.ac.jp.
authors.forename: Mutsumi
authors.initials: M
authors.lastname: Takagi
keywords: Ascorbic acid; Cartilage-like cell sheet; Mesenchymal stem cell; Synergistic effect; Type 2 collagen